TY  - JOUR
AU  - Gligorijević, Nikola
AU  - Jovanović, Zorana
AU  - Cvijetić, Ilija
AU  - Šunderić, Miloš
AU  - Veličković, Luka
AU  - Katrlík, Jaroslav
AU  - Holazová, Alena
AU  - Nikolić, Milan
AU  - Minić, Simeon
PY  - 2024
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7298
AB  - Blue C-phycocyanin (C-PC), the major Spirulina protein with innumerable
health-promoting benefits, is an attractive colourant and food supplement. A crucial obstacle to its
more extensive use is its relatively low stability. This study aimed to screen various food-derived
ligands for their ability to bind and stabilise C-PC, utilising spectroscopic techniques and molecular
docking. Among twelve examined ligands, the protein fluorescence quenching revealed that
only quercetin, coenzyme Q10 and resveratrol had a moderate affinity to C-PC (Ka of 2.2 to 3.7 × 105
M–1). Docking revealed these three ligands bind more strongly to the C-PC hexamer than the trimer,
with the binding sites located at the interface of two (αβ)3 trimers. UV/VIS absorption spectroscopy
demonstrated the changes in the C-PC absorption spectra in a complex with quercetin
and resveratrol compared to the spectra of free protein and ligands. Selected ligands did not affect
the secondary structure content, but they induced changes in the tertiary protein structure in the
CD study. A fluorescence-based thermal stability assay demonstrated quercetin and coenzyme Q10
increased the C-PC melting point by nearly 5 °C. Our study identified food-derived ligands that
interact with C-PC and improve its thermal stability, indicating their potential as stabilising agents
for C-PC in the food industry.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - Investigation of the Potential of Selected Food-Derived Antioxidants to Bind and Stabilise the Bioactive Blue Protein C-Phycocyanin from Cyanobacteria Spirulina
VL  - 25
IS  - 1
SP  - 229
DO  - 10.3390/ijms25010229
ER  - 

@article{
author = "Gligorijević, Nikola and Jovanović, Zorana and Cvijetić, Ilija and Šunderić, Miloš and Veličković, Luka and Katrlík, Jaroslav and Holazová, Alena and Nikolić, Milan and Minić, Simeon",
year = "2024",
abstract = "Blue C-phycocyanin (C-PC), the major Spirulina protein with innumerable
health-promoting benefits, is an attractive colourant and food supplement. A crucial obstacle to its
more extensive use is its relatively low stability. This study aimed to screen various food-derived
ligands for their ability to bind and stabilise C-PC, utilising spectroscopic techniques and molecular
docking. Among twelve examined ligands, the protein fluorescence quenching revealed that
only quercetin, coenzyme Q10 and resveratrol had a moderate affinity to C-PC (Ka of 2.2 to 3.7 × 105
M–1). Docking revealed these three ligands bind more strongly to the C-PC hexamer than the trimer,
with the binding sites located at the interface of two (αβ)3 trimers. UV/VIS absorption spectroscopy
demonstrated the changes in the C-PC absorption spectra in a complex with quercetin
and resveratrol compared to the spectra of free protein and ligands. Selected ligands did not affect
the secondary structure content, but they induced changes in the tertiary protein structure in the
CD study. A fluorescence-based thermal stability assay demonstrated quercetin and coenzyme Q10
increased the C-PC melting point by nearly 5 °C. Our study identified food-derived ligands that
interact with C-PC and improve its thermal stability, indicating their potential as stabilising agents
for C-PC in the food industry.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "Investigation of the Potential of Selected Food-Derived Antioxidants to Bind and Stabilise the Bioactive Blue Protein C-Phycocyanin from Cyanobacteria Spirulina",
volume = "25",
number = "1",
pages = "229",
doi = "10.3390/ijms25010229"
}

Gligorijević, N., Jovanović, Z., Cvijetić, I., Šunderić, M., Veličković, L., Katrlík, J., Holazová, A., Nikolić, M.,& Minić, S.. (2024). Investigation of the Potential of Selected Food-Derived Antioxidants to Bind and Stabilise the Bioactive Blue Protein C-Phycocyanin from Cyanobacteria Spirulina. in International Journal of Molecular Sciences
MDPI., 25(1), 229.
https://doi.org/10.3390/ijms25010229

Gligorijević N, Jovanović Z, Cvijetić I, Šunderić M, Veličković L, Katrlík J, Holazová A, Nikolić M, Minić S. Investigation of the Potential of Selected Food-Derived Antioxidants to Bind and Stabilise the Bioactive Blue Protein C-Phycocyanin from Cyanobacteria Spirulina. in International Journal of Molecular Sciences. 2024;25(1):229.
doi:10.3390/ijms25010229 .

Gligorijević, Nikola, Jovanović, Zorana, Cvijetić, Ilija, Šunderić, Miloš, Veličković, Luka, Katrlík, Jaroslav, Holazová, Alena, Nikolić, Milan, Minić, Simeon, "Investigation of the Potential of Selected Food-Derived Antioxidants to Bind and Stabilise the Bioactive Blue Protein C-Phycocyanin from Cyanobacteria Spirulina" in International Journal of Molecular Sciences, 25, no. 1 (2024):229,
https://doi.org/10.3390/ijms25010229 . .

TY  - CONF
AU  - Jovanović, Zorana
AU  - Annighöfer, Burkhard
AU  - Dudzinski, Daniel
AU  - Gligorijević, Nikola
AU  - Nikolić, Milan
AU  - Pavlović, Vladimir B.
AU  - Lević, Steva
AU  - Brûlet, Annie
AU  - Assifaoui, Ali
AU  - Combet, Sophie
AU  - Minić, Simeon
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7521
AB  - Our study aimed to preserve the natural blue dye of C-phycocyanin (C-PC)
phycobiliprotein from Spirulina microalgae due to its importance in the food industry. We
incorporated C-PC into hydrogels formed by combining starch and β-lactoglobulin (BLG)
using high-pressure (HP) processing to achieve this objective. Notably, thermal treatment
resulted in the complete loss of colour derived from C-PC.
We performed a comprehensive characterization of the resulting HP gels by rheology
measurements, texture profile analysis (TPA), small-angle X-ray scattering (SAXS), and
scanning electron microscopy (SEM).
Different compositions of binary (BLG/C-PC) and ternary (starch/BLG/C-PC)
systems were processed under high-pressure (HP) conditions reaching up to 4,500 bar. The
C-PC pigment was effectively preserved by mixing BLG and starch with C-PC at pH 7,
maintaining concentrations of 180, 5, and 10 mg/mL, respectively. The same concentrations
of components were retained in the binary systems.
Rheological properties of the gels were determined using a rheometer with
plane/plane geometry, and texture analysis was conducted through TPA. These findings
enabled the assessment of food gel's properties, such as hardness, springiness, chewiness, and
cohesiveness. The structural characteristics of the gels were determined by SAXS, offering
insights into the interactions between C-PC, BLG, and starch after HP processing. Adding CPC
and starch formed solid gels with a larger mesh than the pure BLG gels. SEM scans of the
gel surface revealed that all components influenced the overall morphology of gels. Even at
low concentrations, the addition of starch notably influenced the gels' visual appearance and
mechanical properties. Our investigation highlights the superior effectiveness of HP treatment
in the preservation of C-PC compared to high-temperature treatment, evident in the sustained
colour integrity of the C-PC blue dye.
PB  - Faculty of Technology and Metallurgy, University of Belgrade
C3  - Book of abstracts - International Conference on Biochemical Engineering and Biotechnology for Young Scientists, Belgrade
T1  - The use of starch and β-lactoglobulin composite hydrogels as frameworks for preserving c-phycocyanin
SP  - 60
EP  - 60
UR  - https://hdl.handle.net/21.15107/rcub_cer_7521
ER  - 

@conference{
author = "Jovanović, Zorana and Annighöfer, Burkhard and Dudzinski, Daniel and Gligorijević, Nikola and Nikolić, Milan and Pavlović, Vladimir B. and Lević, Steva and Brûlet, Annie and Assifaoui, Ali and Combet, Sophie and Minić, Simeon",
year = "2023",
abstract = "Our study aimed to preserve the natural blue dye of C-phycocyanin (C-PC)
phycobiliprotein from Spirulina microalgae due to its importance in the food industry. We
incorporated C-PC into hydrogels formed by combining starch and β-lactoglobulin (BLG)
using high-pressure (HP) processing to achieve this objective. Notably, thermal treatment
resulted in the complete loss of colour derived from C-PC.
We performed a comprehensive characterization of the resulting HP gels by rheology
measurements, texture profile analysis (TPA), small-angle X-ray scattering (SAXS), and
scanning electron microscopy (SEM).
Different compositions of binary (BLG/C-PC) and ternary (starch/BLG/C-PC)
systems were processed under high-pressure (HP) conditions reaching up to 4,500 bar. The
C-PC pigment was effectively preserved by mixing BLG and starch with C-PC at pH 7,
maintaining concentrations of 180, 5, and 10 mg/mL, respectively. The same concentrations
of components were retained in the binary systems.
Rheological properties of the gels were determined using a rheometer with
plane/plane geometry, and texture analysis was conducted through TPA. These findings
enabled the assessment of food gel's properties, such as hardness, springiness, chewiness, and
cohesiveness. The structural characteristics of the gels were determined by SAXS, offering
insights into the interactions between C-PC, BLG, and starch after HP processing. Adding CPC
and starch formed solid gels with a larger mesh than the pure BLG gels. SEM scans of the
gel surface revealed that all components influenced the overall morphology of gels. Even at
low concentrations, the addition of starch notably influenced the gels' visual appearance and
mechanical properties. Our investigation highlights the superior effectiveness of HP treatment
in the preservation of C-PC compared to high-temperature treatment, evident in the sustained
colour integrity of the C-PC blue dye.",
publisher = "Faculty of Technology and Metallurgy, University of Belgrade",
journal = "Book of abstracts - International Conference on Biochemical Engineering and Biotechnology for Young Scientists, Belgrade",
title = "The use of starch and β-lactoglobulin composite hydrogels as frameworks for preserving c-phycocyanin",
pages = "60-60",
url = "https://hdl.handle.net/21.15107/rcub_cer_7521"
}

Jovanović, Z., Annighöfer, B., Dudzinski, D., Gligorijević, N., Nikolić, M., Pavlović, V. B., Lević, S., Brûlet, A., Assifaoui, A., Combet, S.,& Minić, S.. (2023). The use of starch and β-lactoglobulin composite hydrogels as frameworks for preserving c-phycocyanin. in Book of abstracts - International Conference on Biochemical Engineering and Biotechnology for Young Scientists, Belgrade
Faculty of Technology and Metallurgy, University of Belgrade., 60-60.
https://hdl.handle.net/21.15107/rcub_cer_7521

Jovanović Z, Annighöfer B, Dudzinski D, Gligorijević N, Nikolić M, Pavlović VB, Lević S, Brûlet A, Assifaoui A, Combet S, Minić S. The use of starch and β-lactoglobulin composite hydrogels as frameworks for preserving c-phycocyanin. in Book of abstracts - International Conference on Biochemical Engineering and Biotechnology for Young Scientists, Belgrade. 2023;:60-60.
https://hdl.handle.net/21.15107/rcub_cer_7521 .

Jovanović, Zorana, Annighöfer, Burkhard, Dudzinski, Daniel, Gligorijević, Nikola, Nikolić, Milan, Pavlović, Vladimir B., Lević, Steva, Brûlet, Annie, Assifaoui, Ali, Combet, Sophie, Minić, Simeon, "The use of starch and β-lactoglobulin composite hydrogels as frameworks for preserving c-phycocyanin" in Book of abstracts - International Conference on Biochemical Engineering and Biotechnology for Young Scientists, Belgrade (2023):60-60,
https://hdl.handle.net/21.15107/rcub_cer_7521 .

TY  - JOUR
AU  - Šunderić, Miloš
AU  - Gligorijević, Nikola
AU  - Milčić, Miloš
AU  - Minić, Simeon
AU  - Nedić, Olgica
AU  - Nikolić, Milan
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6471
AB  - Under simulated physiological conditions, this study investigates the interaction between nutraceutical phycocyanobilin (PCB) and the universal anti-protease protein human alpha-2-macroglobulin (a2M). Extensive molecular docking analyses on multiple a2M conformations, spectroscopic techniques, and a2M activity assays were utilized to examine the complex formation. The results revealed that for every protein conformation, two high energy binding sites exist: the first, conformationally independent, at the interface region between two monomer chains and the second, conformationally dependent, in the pocket composed of amino acids from four distinct domains (TED, RBD, CUB, and MG2) of the single protein chain. Spectrofluorimetric measurements indicated a moderate affinity between a2M and PCB with a moderately high binding constant of 6.3 x 10^5 M^-1 at 25 °C. The binding of PCB to a2M resulted in minor changes in the secondary structure content of a2M. Furthermore, PCB protected a2M from oxidation and preserved its anti-protease activity in the oxidative environment. These findings suggest that PCB binding could indirectly impact the body’s response to oxidative stress by influencing a2M’s role in controlling enzyme activity during the inflammatory process.
PB  - Taylor & Francis Group
T2  - Journal of Biomolecular Structure and Dynamics
T1  - Phycocyanobilin is a new binding partner of human alpha-2-macroglobulin that protects the protein against oxidative stress
DO  - 10.1080/07391102.2023.2248273
ER  - 

@article{
author = "Šunderić, Miloš and Gligorijević, Nikola and Milčić, Miloš and Minić, Simeon and Nedić, Olgica and Nikolić, Milan",
year = "2023",
abstract = "Under simulated physiological conditions, this study investigates the interaction between nutraceutical phycocyanobilin (PCB) and the universal anti-protease protein human alpha-2-macroglobulin (a2M). Extensive molecular docking analyses on multiple a2M conformations, spectroscopic techniques, and a2M activity assays were utilized to examine the complex formation. The results revealed that for every protein conformation, two high energy binding sites exist: the first, conformationally independent, at the interface region between two monomer chains and the second, conformationally dependent, in the pocket composed of amino acids from four distinct domains (TED, RBD, CUB, and MG2) of the single protein chain. Spectrofluorimetric measurements indicated a moderate affinity between a2M and PCB with a moderately high binding constant of 6.3 x 10^5 M^-1 at 25 °C. The binding of PCB to a2M resulted in minor changes in the secondary structure content of a2M. Furthermore, PCB protected a2M from oxidation and preserved its anti-protease activity in the oxidative environment. These findings suggest that PCB binding could indirectly impact the body’s response to oxidative stress by influencing a2M’s role in controlling enzyme activity during the inflammatory process.",
publisher = "Taylor & Francis Group",
journal = "Journal of Biomolecular Structure and Dynamics",
title = "Phycocyanobilin is a new binding partner of human alpha-2-macroglobulin that protects the protein against oxidative stress",
doi = "10.1080/07391102.2023.2248273"
}

Šunderić, M., Gligorijević, N., Milčić, M., Minić, S., Nedić, O.,& Nikolić, M.. (2023). Phycocyanobilin is a new binding partner of human alpha-2-macroglobulin that protects the protein against oxidative stress. in Journal of Biomolecular Structure and Dynamics
Taylor & Francis Group..
https://doi.org/10.1080/07391102.2023.2248273

Šunderić M, Gligorijević N, Milčić M, Minić S, Nedić O, Nikolić M. Phycocyanobilin is a new binding partner of human alpha-2-macroglobulin that protects the protein against oxidative stress. in Journal of Biomolecular Structure and Dynamics. 2023;.
doi:10.1080/07391102.2023.2248273 .

Šunderić, Miloš, Gligorijević, Nikola, Milčić, Miloš, Minić, Simeon, Nedić, Olgica, Nikolić, Milan, "Phycocyanobilin is a new binding partner of human alpha-2-macroglobulin that protects the protein against oxidative stress" in Journal of Biomolecular Structure and Dynamics (2023),
https://doi.org/10.1080/07391102.2023.2248273 . .

TY  - JOUR
AU  - Veličković, Luka
AU  - Simović, Ana
AU  - Gligorijević, Nikola
AU  - Thureau, Aurelien
AU  - Obradović, Milica
AU  - Vasović, Tamara
AU  - Sotiroudis, Georgios
AU  - Zoumpanioti, Maria
AU  - Brulet, Annie
AU  - Ćirković Veličković, Tanja
AU  - Combet, Sophie
AU  - Nikolić, Milan
AU  - Minić, Simeon
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6473
AB  - This study aimed to purify, characterise and stabilise the natural food colourant, R-phycocyanin (R-PC), from the red algae Porphyra spp. (Nori). We purified R-PC from dried Nori flakes with a high purity ratio (A618/A280 ≥ 3.4) in native form (α-helix content 53%). SAXS measurements revealed that R-PC is trimeric ((αβ)3) in solution. The thermal denaturation of α-helix revealed one transition (Tm at 52 ◦C), while the pH stability study showed R-PC is stable in the pH range 4–8. The thermal treatment of R-PC at 60 °C has detrimental and irreversible effects on RPC colour and antioxidant capacity (22 % of residual capacity). However, immobilisation of R-PC within calcium alginate beads completely preserves R-PC colour and mainly retains its antioxidant ability (78 % of residual
capacity). Results give new insights into the stability of R-PC and preservation of its purple colour and bioactivity by encapsulation in calcium alginate beads.
PB  - Elsevier Ltd.
T2  - Food Chemistry
T1  - Exploring and strengthening the potential of R-phycocyanin from Nori flakes as a food colourant
VL  - 426
SP  - 136669
DO  - 10.1016/j.foodchem.2023.136669
ER  - 

@article{
author = "Veličković, Luka and Simović, Ana and Gligorijević, Nikola and Thureau, Aurelien and Obradović, Milica and Vasović, Tamara and Sotiroudis, Georgios and Zoumpanioti, Maria and Brulet, Annie and Ćirković Veličković, Tanja and Combet, Sophie and Nikolić, Milan and Minić, Simeon",
year = "2023",
abstract = "This study aimed to purify, characterise and stabilise the natural food colourant, R-phycocyanin (R-PC), from the red algae Porphyra spp. (Nori). We purified R-PC from dried Nori flakes with a high purity ratio (A618/A280 ≥ 3.4) in native form (α-helix content 53%). SAXS measurements revealed that R-PC is trimeric ((αβ)3) in solution. The thermal denaturation of α-helix revealed one transition (Tm at 52 ◦C), while the pH stability study showed R-PC is stable in the pH range 4–8. The thermal treatment of R-PC at 60 °C has detrimental and irreversible effects on RPC colour and antioxidant capacity (22 % of residual capacity). However, immobilisation of R-PC within calcium alginate beads completely preserves R-PC colour and mainly retains its antioxidant ability (78 % of residual
capacity). Results give new insights into the stability of R-PC and preservation of its purple colour and bioactivity by encapsulation in calcium alginate beads.",
publisher = "Elsevier Ltd.",
journal = "Food Chemistry",
title = "Exploring and strengthening the potential of R-phycocyanin from Nori flakes as a food colourant",
volume = "426",
pages = "136669",
doi = "10.1016/j.foodchem.2023.136669"
}

Veličković, L., Simović, A., Gligorijević, N., Thureau, A., Obradović, M., Vasović, T., Sotiroudis, G., Zoumpanioti, M., Brulet, A., Ćirković Veličković, T., Combet, S., Nikolić, M.,& Minić, S.. (2023). Exploring and strengthening the potential of R-phycocyanin from Nori flakes as a food colourant. in Food Chemistry
Elsevier Ltd.., 426, 136669.
https://doi.org/10.1016/j.foodchem.2023.136669

Veličković L, Simović A, Gligorijević N, Thureau A, Obradović M, Vasović T, Sotiroudis G, Zoumpanioti M, Brulet A, Ćirković Veličković T, Combet S, Nikolić M, Minić S. Exploring and strengthening the potential of R-phycocyanin from Nori flakes as a food colourant. in Food Chemistry. 2023;426:136669.
doi:10.1016/j.foodchem.2023.136669 .

Veličković, Luka, Simović, Ana, Gligorijević, Nikola, Thureau, Aurelien, Obradović, Milica, Vasović, Tamara, Sotiroudis, Georgios, Zoumpanioti, Maria, Brulet, Annie, Ćirković Veličković, Tanja, Combet, Sophie, Nikolić, Milan, Minić, Simeon, "Exploring and strengthening the potential of R-phycocyanin from Nori flakes as a food colourant" in Food Chemistry, 426 (2023):136669,
https://doi.org/10.1016/j.foodchem.2023.136669 . .

TY  - JOUR
AU  - Platanić Arizanović, Lena
AU  - Gligorijević, Nikola
AU  - Cvijetić, Ilija
AU  - Mijatović, Aleksandar
AU  - Krstić Ristivojević, Maja
AU  - Minić, Simeon
AU  - Nikolić Kokić, Aleksandra
AU  - Miljević, Čedo
AU  - Nikolić, Milan
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6482
AB  - Packed with hemoglobin, an essential protein for oxygen transport, human erythrocytes
are a suitable model system for testing the pleiotropic effects of lipophilic drugs. Our study investigated the interaction between antipsychotic drugs clozapine, ziprasidone, sertindole, and human hemoglobin under simulated physiological conditions. Analysis of protein fluorescence quenching at different temperatures and data obtained from the van’t Hoff diagram and molecular docking indicate that the interactions are static and that the tetrameric human hemoglobin has one binding site for all drugs in the central cavity near αβ  interfaces and is dominantly mediated through hydrophobic forces. The association constants were lower-moderate strength (~10^4 M^-1), the highest observed for clozapine (2.2 x 10^4 M^-1 at 25 °C). The clozapine binding showed “friendly” effects: increased α-helical content, a higher melting point, and protein protection from free radical-mediated oxidation. On the other hand, bound ziprasidone and sertindole had a slightly pro-oxidative effect, increasing ferrihemoglobin content, a possible “foe”. Since the interaction of proteins with drugs plays a vital role in their pharmacokinetic and pharmacodynamic properties, the physiological significance of the obtained findings is briefly discussed.
PB  - Multidisciplinary Digital Publishing Institute (MDPI)
T2  - International Journal of Molecular Sciences
T1  - Human Hemoglobin and Antipsychotics Clozapine, Ziprasidone and Sertindole: Friends or Foes?
VL  - 24
IS  - 10
SP  - 8921
DO  - 10.3390/ijms24108921
ER  - 

@article{
author = "Platanić Arizanović, Lena and Gligorijević, Nikola and Cvijetić, Ilija and Mijatović, Aleksandar and Krstić Ristivojević, Maja and Minić, Simeon and Nikolić Kokić, Aleksandra and Miljević, Čedo and Nikolić, Milan",
year = "2023",
abstract = "Packed with hemoglobin, an essential protein for oxygen transport, human erythrocytes
are a suitable model system for testing the pleiotropic effects of lipophilic drugs. Our study investigated the interaction between antipsychotic drugs clozapine, ziprasidone, sertindole, and human hemoglobin under simulated physiological conditions. Analysis of protein fluorescence quenching at different temperatures and data obtained from the van’t Hoff diagram and molecular docking indicate that the interactions are static and that the tetrameric human hemoglobin has one binding site for all drugs in the central cavity near αβ  interfaces and is dominantly mediated through hydrophobic forces. The association constants were lower-moderate strength (~10^4 M^-1), the highest observed for clozapine (2.2 x 10^4 M^-1 at 25 °C). The clozapine binding showed “friendly” effects: increased α-helical content, a higher melting point, and protein protection from free radical-mediated oxidation. On the other hand, bound ziprasidone and sertindole had a slightly pro-oxidative effect, increasing ferrihemoglobin content, a possible “foe”. Since the interaction of proteins with drugs plays a vital role in their pharmacokinetic and pharmacodynamic properties, the physiological significance of the obtained findings is briefly discussed.",
publisher = "Multidisciplinary Digital Publishing Institute (MDPI)",
journal = "International Journal of Molecular Sciences",
title = "Human Hemoglobin and Antipsychotics Clozapine, Ziprasidone and Sertindole: Friends or Foes?",
volume = "24",
number = "10",
pages = "8921",
doi = "10.3390/ijms24108921"
}

Platanić Arizanović, L., Gligorijević, N., Cvijetić, I., Mijatović, A., Krstić Ristivojević, M., Minić, S., Nikolić Kokić, A., Miljević, Č.,& Nikolić, M.. (2023). Human Hemoglobin and Antipsychotics Clozapine, Ziprasidone and Sertindole: Friends or Foes?. in International Journal of Molecular Sciences
Multidisciplinary Digital Publishing Institute (MDPI)., 24(10), 8921.
https://doi.org/10.3390/ijms24108921

Platanić Arizanović L, Gligorijević N, Cvijetić I, Mijatović A, Krstić Ristivojević M, Minić S, Nikolić Kokić A, Miljević Č, Nikolić M. Human Hemoglobin and Antipsychotics Clozapine, Ziprasidone and Sertindole: Friends or Foes?. in International Journal of Molecular Sciences. 2023;24(10):8921.
doi:10.3390/ijms24108921 .

Platanić Arizanović, Lena, Gligorijević, Nikola, Cvijetić, Ilija, Mijatović, Aleksandar, Krstić Ristivojević, Maja, Minić, Simeon, Nikolić Kokić, Aleksandra, Miljević, Čedo, Nikolić, Milan, "Human Hemoglobin and Antipsychotics Clozapine, Ziprasidone and Sertindole: Friends or Foes?" in International Journal of Molecular Sciences, 24, no. 10 (2023):8921,
https://doi.org/10.3390/ijms24108921 . .

TY  - JOUR
AU  - Nedić, Olgica
AU  - Penezić, Ana
AU  - Minić, Simeon
AU  - Radomirović, Mirjana
AU  - Nikolić, Milan
AU  - Ćirković Veličković, Tanja
AU  - Gligorijević, Nikola
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6485
AB  - Common to all biological systems and living organisms are molecular interactions, which
may lead to specific physiological events. Most often, a cascade of events occurs, establishing an
equilibrium between possibly competing and/or synergistic processes. Biochemical pathways that
sustain life depend on multiple intrinsic and extrinsic factors contributing to aging and/or diseases.
This article deals with food antioxidants and human proteins from the circulation, their interaction,
their effect on the structure, properties, and function of antioxidant-bound proteins, and the possible
impact of complex formation on antioxidants. An overview of studies examining interactions between
individual antioxidant compounds and major blood proteins is presented with findings. Investigating
antioxidant/protein interactions at the level of the human organism and determining antioxidant
distribution between proteins and involvement in the particular physiological role is a very complex
and challenging task. However, by knowing the role of a particular protein in certain pathology or
aging, and the effect exerted by a particular antioxidant bound to it, it is possible to recommend
specific food intake or resistance to it to improve the condition or slow down the process.
PB  - Multidisciplinary Digital Publishing Institute (MDPI)
T2  - Antioxidants
T1  - Food Antioxidants and Their Interaction with Human Proteins
VL  - 12
IS  - 4
SP  - 815
DO  - 10.3390/antiox12040815
ER  - 

@article{
author = "Nedić, Olgica and Penezić, Ana and Minić, Simeon and Radomirović, Mirjana and Nikolić, Milan and Ćirković Veličković, Tanja and Gligorijević, Nikola",
year = "2023",
abstract = "Common to all biological systems and living organisms are molecular interactions, which
may lead to specific physiological events. Most often, a cascade of events occurs, establishing an
equilibrium between possibly competing and/or synergistic processes. Biochemical pathways that
sustain life depend on multiple intrinsic and extrinsic factors contributing to aging and/or diseases.
This article deals with food antioxidants and human proteins from the circulation, their interaction,
their effect on the structure, properties, and function of antioxidant-bound proteins, and the possible
impact of complex formation on antioxidants. An overview of studies examining interactions between
individual antioxidant compounds and major blood proteins is presented with findings. Investigating
antioxidant/protein interactions at the level of the human organism and determining antioxidant
distribution between proteins and involvement in the particular physiological role is a very complex
and challenging task. However, by knowing the role of a particular protein in certain pathology or
aging, and the effect exerted by a particular antioxidant bound to it, it is possible to recommend
specific food intake or resistance to it to improve the condition or slow down the process.",
publisher = "Multidisciplinary Digital Publishing Institute (MDPI)",
journal = "Antioxidants",
title = "Food Antioxidants and Their Interaction with Human Proteins",
volume = "12",
number = "4",
pages = "815",
doi = "10.3390/antiox12040815"
}

Nedić, O., Penezić, A., Minić, S., Radomirović, M., Nikolić, M., Ćirković Veličković, T.,& Gligorijević, N.. (2023). Food Antioxidants and Their Interaction with Human Proteins. in Antioxidants
Multidisciplinary Digital Publishing Institute (MDPI)., 12(4), 815.
https://doi.org/10.3390/antiox12040815

Nedić O, Penezić A, Minić S, Radomirović M, Nikolić M, Ćirković Veličković T, Gligorijević N. Food Antioxidants and Their Interaction with Human Proteins. in Antioxidants. 2023;12(4):815.
doi:10.3390/antiox12040815 .

Nedić, Olgica, Penezić, Ana, Minić, Simeon, Radomirović, Mirjana, Nikolić, Milan, Ćirković Veličković, Tanja, Gligorijević, Nikola, "Food Antioxidants and Their Interaction with Human Proteins" in Antioxidants, 12, no. 4 (2023):815,
https://doi.org/10.3390/antiox12040815 . .

TY  - CONF
AU  - Veličković, Luka
AU  - Simović, Ana
AU  - Gligorijević, Nikola
AU  - Obradović, Milica
AU  - Sotiroudis, Georgios
AU  - Zoumpanioti, Maria
AU  - Minić, Simeon
AU  - Nikolić, Milan
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6505
AB  - Purple R-phycocyanin is a protein from red algae with the potential for application in the food industry (colorant) and wastewater treatment (binding of heavy metals). Analytical grade R-phycocyanin was purified from the buffered extract of dried Nori flakes (Porphyra spp.) by combining ammonium sulfate precipitation, hydroxyapatite, and DEAE-Sepharose column chromatography. The multimeric protein had absorption maxima characteristic for phycoerythrobilin (at 580 nm) and phycocyanobilin (at 640 nm) chromophores, high α- helical content, and melting temperature of 52°C. The secondary R-PC structure was stable under a wide range of pH values (3–9). R-phycocyanin immobilized in calcium alginate beads showed increased thermal stability and preserved antioxidant activity.
AB  - Ljubičasti R-fikocijanin je protein crvenih algi sa mogućnostima primene u industriji hrane (kolorant) i za tretman otpadnih voda (vezuje teške metale). R-fikocijanin analitičke čistoće je izolovan iz puferisanog ekstrakta osušenih Nori algi (Porphyra spp.), kombinacijom taloženja amonijum-sulfatom, hidroksiapatitne i hromatografije na DEAE-Sepharose koloni. Multimerni protein imao je apsorpcione maksimume karakteristične za fikoeritrobilinsku (na 580 nm) i fikocijanobilinsku (na 640 nm) hromoforu, visok sadržaj α-zavojnica i temperaturu topljenja od 52°C. Sekundarna struktura proteina bila je stabilna u širokom rasponu pH vrednosti (3–9). R-fikocijanin imobilisan u kuglice kalcijum-alginata pokazao je povećanu toplotnu stabilnost i očuvana antioksidativna svojstva.
PB  - Belgrade : Serbian Chemical Society
C3  - Kratki izvodi radova, knjiga radova - 59. Savetovanje Srpskog hemijskog društva, 1. i 2. jun 2023. godine, Novi Sad / Book of Abstracts, Proceedings - 59th Meeting of the Serbian Chemical Society, June 1-2, 2023, Novi Sad, Serbia
T1  - Purification and structural characterization of R-phycocyanin
T1  - Prečišćavanje i strukturna karakterizacija R-fikocijanina
SP  - 53
EP  - 53
UR  - https://hdl.handle.net/21.15107/rcub_cer_6505
ER  - 

@conference{
author = "Veličković, Luka and Simović, Ana and Gligorijević, Nikola and Obradović, Milica and Sotiroudis, Georgios and Zoumpanioti, Maria and Minić, Simeon and Nikolić, Milan",
year = "2023",
abstract = "Purple R-phycocyanin is a protein from red algae with the potential for application in the food industry (colorant) and wastewater treatment (binding of heavy metals). Analytical grade R-phycocyanin was purified from the buffered extract of dried Nori flakes (Porphyra spp.) by combining ammonium sulfate precipitation, hydroxyapatite, and DEAE-Sepharose column chromatography. The multimeric protein had absorption maxima characteristic for phycoerythrobilin (at 580 nm) and phycocyanobilin (at 640 nm) chromophores, high α- helical content, and melting temperature of 52°C. The secondary R-PC structure was stable under a wide range of pH values (3–9). R-phycocyanin immobilized in calcium alginate beads showed increased thermal stability and preserved antioxidant activity., Ljubičasti R-fikocijanin je protein crvenih algi sa mogućnostima primene u industriji hrane (kolorant) i za tretman otpadnih voda (vezuje teške metale). R-fikocijanin analitičke čistoće je izolovan iz puferisanog ekstrakta osušenih Nori algi (Porphyra spp.), kombinacijom taloženja amonijum-sulfatom, hidroksiapatitne i hromatografije na DEAE-Sepharose koloni. Multimerni protein imao je apsorpcione maksimume karakteristične za fikoeritrobilinsku (na 580 nm) i fikocijanobilinsku (na 640 nm) hromoforu, visok sadržaj α-zavojnica i temperaturu topljenja od 52°C. Sekundarna struktura proteina bila je stabilna u širokom rasponu pH vrednosti (3–9). R-fikocijanin imobilisan u kuglice kalcijum-alginata pokazao je povećanu toplotnu stabilnost i očuvana antioksidativna svojstva.",
publisher = "Belgrade : Serbian Chemical Society",
journal = "Kratki izvodi radova, knjiga radova - 59. Savetovanje Srpskog hemijskog društva, 1. i 2. jun 2023. godine, Novi Sad / Book of Abstracts, Proceedings - 59th Meeting of the Serbian Chemical Society, June 1-2, 2023, Novi Sad, Serbia",
title = "Purification and structural characterization of R-phycocyanin, Prečišćavanje i strukturna karakterizacija R-fikocijanina",
pages = "53-53",
url = "https://hdl.handle.net/21.15107/rcub_cer_6505"
}

Veličković, L., Simović, A., Gligorijević, N., Obradović, M., Sotiroudis, G., Zoumpanioti, M., Minić, S.,& Nikolić, M.. (2023). Purification and structural characterization of R-phycocyanin. in Kratki izvodi radova, knjiga radova - 59. Savetovanje Srpskog hemijskog društva, 1. i 2. jun 2023. godine, Novi Sad / Book of Abstracts, Proceedings - 59th Meeting of the Serbian Chemical Society, June 1-2, 2023, Novi Sad, Serbia
Belgrade : Serbian Chemical Society., 53-53.
https://hdl.handle.net/21.15107/rcub_cer_6505

Veličković L, Simović A, Gligorijević N, Obradović M, Sotiroudis G, Zoumpanioti M, Minić S, Nikolić M. Purification and structural characterization of R-phycocyanin. in Kratki izvodi radova, knjiga radova - 59. Savetovanje Srpskog hemijskog društva, 1. i 2. jun 2023. godine, Novi Sad / Book of Abstracts, Proceedings - 59th Meeting of the Serbian Chemical Society, June 1-2, 2023, Novi Sad, Serbia. 2023;:53-53.
https://hdl.handle.net/21.15107/rcub_cer_6505 .

Veličković, Luka, Simović, Ana, Gligorijević, Nikola, Obradović, Milica, Sotiroudis, Georgios, Zoumpanioti, Maria, Minić, Simeon, Nikolić, Milan, "Purification and structural characterization of R-phycocyanin" in Kratki izvodi radova, knjiga radova - 59. Savetovanje Srpskog hemijskog društva, 1. i 2. jun 2023. godine, Novi Sad / Book of Abstracts, Proceedings - 59th Meeting of the Serbian Chemical Society, June 1-2, 2023, Novi Sad, Serbia (2023):53-53,
https://hdl.handle.net/21.15107/rcub_cer_6505 .

TY  - CONF
AU  - Gligorijević, Nikola
AU  - Veličković, Luka
AU  - Svrzić, Nikola
AU  - Jovanović, Zorana
AU  - Minić, Simeon
AU  - Nikolić, Milan
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6519
AB  - C-Phycocyanin (C-PC), the major protein of cyanobacteria Arthrospira platensis, is a phycobiliprotein with potent biological activity. It has several beneficial effects, including anti-oxidant, anti-inflammatory, immunomodulatory, and anti-cancer. A significant challenge for the broader application of C-PC in the food industry is its stability in food processing conditions, such
as increased light exposure, temperature, and high pressure and drying. This work aimed to investigate if the immobilization of C-PC onto alginate beads could improve its stability. C-PC was
immobilized by dropping the solution of C-PC and 1% alginate (final concentration) in the solution of 2% CaCl2. Both protein/alginate mixture and CaCl2 were kept at pH 4. Immobilized CPC
was treated for 30 min at 65°C, by high pressure up to 4500 bar, and incubated under light exposure for a month. Alginate beads with immobilized C-PC were also left to dry in the fridge and kept for a month. C-PC was extracted from alginate beads by immersing them in 20 mM phosphate buffer, pH 7. The stability of C-PC was assessed by a color change and UV-VIS spectroscopy. Immobilized C-PC was stable under all tested conditions, with only small aggregation and color change appearing
after high-pressure treatment. Immobilization of C-PC by alginate thus shows promise for its efficient stabilization under food processing conditions.
PB  - Wiley
C3  - FEBS Open Bio
T1  - Stabilization of C-phycocyanin by immobilization in alginate beads
VL  - 13
IS  - S2
SP  - 234
EP  - 234
DO  - 10.1002/2211-5463.13646
ER  - 

@conference{
author = "Gligorijević, Nikola and Veličković, Luka and Svrzić, Nikola and Jovanović, Zorana and Minić, Simeon and Nikolić, Milan",
year = "2023",
abstract = "C-Phycocyanin (C-PC), the major protein of cyanobacteria Arthrospira platensis, is a phycobiliprotein with potent biological activity. It has several beneficial effects, including anti-oxidant, anti-inflammatory, immunomodulatory, and anti-cancer. A significant challenge for the broader application of C-PC in the food industry is its stability in food processing conditions, such
as increased light exposure, temperature, and high pressure and drying. This work aimed to investigate if the immobilization of C-PC onto alginate beads could improve its stability. C-PC was
immobilized by dropping the solution of C-PC and 1% alginate (final concentration) in the solution of 2% CaCl2. Both protein/alginate mixture and CaCl2 were kept at pH 4. Immobilized CPC
was treated for 30 min at 65°C, by high pressure up to 4500 bar, and incubated under light exposure for a month. Alginate beads with immobilized C-PC were also left to dry in the fridge and kept for a month. C-PC was extracted from alginate beads by immersing them in 20 mM phosphate buffer, pH 7. The stability of C-PC was assessed by a color change and UV-VIS spectroscopy. Immobilized C-PC was stable under all tested conditions, with only small aggregation and color change appearing
after high-pressure treatment. Immobilization of C-PC by alginate thus shows promise for its efficient stabilization under food processing conditions.",
publisher = "Wiley",
journal = "FEBS Open Bio",
title = "Stabilization of C-phycocyanin by immobilization in alginate beads",
volume = "13",
number = "S2",
pages = "234-234",
doi = "10.1002/2211-5463.13646"
}

Gligorijević, N., Veličković, L., Svrzić, N., Jovanović, Z., Minić, S.,& Nikolić, M.. (2023). Stabilization of C-phycocyanin by immobilization in alginate beads. in FEBS Open Bio
Wiley., 13(S2), 234-234.
https://doi.org/10.1002/2211-5463.13646

Gligorijević N, Veličković L, Svrzić N, Jovanović Z, Minić S, Nikolić M. Stabilization of C-phycocyanin by immobilization in alginate beads. in FEBS Open Bio. 2023;13(S2):234-234.
doi:10.1002/2211-5463.13646 .

Gligorijević, Nikola, Veličković, Luka, Svrzić, Nikola, Jovanović, Zorana, Minić, Simeon, Nikolić, Milan, "Stabilization of C-phycocyanin by immobilization in alginate beads" in FEBS Open Bio, 13, no. S2 (2023):234-234,
https://doi.org/10.1002/2211-5463.13646 . .

TY  - CONF
AU  - Šunderić, Miloš
AU  - Veličković, Luka
AU  - Gligorijević, Nikola
AU  - Aleksić, Ljubodrag
AU  - Nikolić, Milan
AU  - Takić, Marija
AU  - Minić, Simeon
PY  - 2023
UR  - https://euroanalysis2023.ch/
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6592
AB  - C-Phycocyanin (C-PC) is a phycobiliprotein from cyanobacteria, where it harvests light energy that is then transferred to chlorophylls during photosynthesis. It has an intense blue color due to a covalently bonded tetrapyrrole chromophore, and owing to this property is used in the food industry as a good natural alternative for food coloring. In addition to its coloring properties, C-PC has anti-inflammatory, antioxidant, anti-cancer, and immune-enhancing effects that qualify it as a dietary supplement already included in various formulations, mainly Spirulina extract powders. Since it is used as a food colorant and as a dietary supplement, it may interact with food ingredients, affecting its stability, digestibility, or antioxidant properties. Palmitic acid and linoleic acid (which can be metabolized to linolenic acid) are abundant in meat, milk, and edible oils, so that they could interact with C-PC. C-Phycocyanin isolated from the cyanobacterium Arthrospira platensis (Spirulina) was incubated with increasing concentrations of these three fatty acids, and its fluorescence intensity was monitored. Incubation resulted in a fluorescence quenching effect, indicating that binding had occurred. The binding equations indicated that the association constants were of the same order of magnitude and that the number of approximate binding sites was more than one (Ka = 4.64 x 10⁴ M-¹, n = 1.5 for linoleic acid; Ka = 2.88 x 10⁴ M–¹, n = 1.9 for linolenic acid; Ka = 0.44 x 10⁴ M–¹, n = 0.8 for palmitic acid). This moderate interaction between C-PC and fatty acids could influence its behavior as a nutraceutical and food colorant.
PB  - European Chemical Society
C3  - Euroanalysis 2023 - Analytical Probing of Complex Systems, Abstract book, August 27th - 31st, 2023, Geneva, Switzerland
T1  - Dietary fatty acids as a new binding partner of C - phycocyanin: a fluorimetric study
SP  - 332
EP  - 333
UR  - https://hdl.handle.net/21.15107/rcub_cer_6592
ER  - 

@conference{
author = "Šunderić, Miloš and Veličković, Luka and Gligorijević, Nikola and Aleksić, Ljubodrag and Nikolić, Milan and Takić, Marija and Minić, Simeon",
year = "2023",
abstract = "C-Phycocyanin (C-PC) is a phycobiliprotein from cyanobacteria, where it harvests light energy that is then transferred to chlorophylls during photosynthesis. It has an intense blue color due to a covalently bonded tetrapyrrole chromophore, and owing to this property is used in the food industry as a good natural alternative for food coloring. In addition to its coloring properties, C-PC has anti-inflammatory, antioxidant, anti-cancer, and immune-enhancing effects that qualify it as a dietary supplement already included in various formulations, mainly Spirulina extract powders. Since it is used as a food colorant and as a dietary supplement, it may interact with food ingredients, affecting its stability, digestibility, or antioxidant properties. Palmitic acid and linoleic acid (which can be metabolized to linolenic acid) are abundant in meat, milk, and edible oils, so that they could interact with C-PC. C-Phycocyanin isolated from the cyanobacterium Arthrospira platensis (Spirulina) was incubated with increasing concentrations of these three fatty acids, and its fluorescence intensity was monitored. Incubation resulted in a fluorescence quenching effect, indicating that binding had occurred. The binding equations indicated that the association constants were of the same order of magnitude and that the number of approximate binding sites was more than one (Ka = 4.64 x 10⁴ M-¹, n = 1.5 for linoleic acid; Ka = 2.88 x 10⁴ M–¹, n = 1.9 for linolenic acid; Ka = 0.44 x 10⁴ M–¹, n = 0.8 for palmitic acid). This moderate interaction between C-PC and fatty acids could influence its behavior as a nutraceutical and food colorant.",
publisher = "European Chemical Society",
journal = "Euroanalysis 2023 - Analytical Probing of Complex Systems, Abstract book, August 27th - 31st, 2023, Geneva, Switzerland",
title = "Dietary fatty acids as a new binding partner of C - phycocyanin: a fluorimetric study",
pages = "332-333",
url = "https://hdl.handle.net/21.15107/rcub_cer_6592"
}

Šunderić, M., Veličković, L., Gligorijević, N., Aleksić, L., Nikolić, M., Takić, M.,& Minić, S.. (2023). Dietary fatty acids as a new binding partner of C - phycocyanin: a fluorimetric study. in Euroanalysis 2023 - Analytical Probing of Complex Systems, Abstract book, August 27th - 31st, 2023, Geneva, Switzerland
European Chemical Society., 332-333.
https://hdl.handle.net/21.15107/rcub_cer_6592

Šunderić M, Veličković L, Gligorijević N, Aleksić L, Nikolić M, Takić M, Minić S. Dietary fatty acids as a new binding partner of C - phycocyanin: a fluorimetric study. in Euroanalysis 2023 - Analytical Probing of Complex Systems, Abstract book, August 27th - 31st, 2023, Geneva, Switzerland. 2023;:332-333.
https://hdl.handle.net/21.15107/rcub_cer_6592 .

Šunderić, Miloš, Veličković, Luka, Gligorijević, Nikola, Aleksić, Ljubodrag, Nikolić, Milan, Takić, Marija, Minić, Simeon, "Dietary fatty acids as a new binding partner of C - phycocyanin: a fluorimetric study" in Euroanalysis 2023 - Analytical Probing of Complex Systems, Abstract book, August 27th - 31st, 2023, Geneva, Switzerland (2023):332-333,
https://hdl.handle.net/21.15107/rcub_cer_6592 .

TY  - CONF
AU  - Ivanov, Aleksandar
AU  - Veličković, Luka
AU  - Jovanović, Zorana
AU  - Gligorijević, Nikola
AU  - Minić, Simeon
AU  - Nikolić, Milan
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6665
AB  - C-phycocyanin (C-PC) is a photosynthetic protein from Arthrospira platensis
(cyanobacteria). Due to its intense blue colour, which is very rare in nature, C-PC has
industrial applications as a food colourant as a substitute for synthetic food colourants.
Disadvantages of C-PC as a food colourant are its poor stability at high temperatures
(during thermal treatment of the food) and its sensibility to change pH value. The binding
of food-derived small molecules, such as vitamins, could stabilize the structure of C-PC at
high temperatures and wide pH ranges. In this study, we characterized the binding of
selected vitamins to C-PC, purified from the commercial powder of Arthrospira platensis.
We used hydrophilic vitamins (B1, B2, B7, B9, B12), lipophilic vitamins (A, D3) and
provitamin (β-carotene). Fluorescent spectroscopy showed a decrease in fluorescence of CPC
i n t he p resence o f v itamin A, v itamin D3 a nd β -carotene (lipophilic molecules)
compared to the control. In contrast, the fluorescence of C-PC in the presence of
hydrophilic vitamins showed minimal change. The protein fluorescence quenching
approach demonstrated hydrophobic (pro)vitamins binding affinities ranging from 0.02 to
5.9 x 105 M-1, with the ability of hydrophobic (pro)vitamins to bind at the different sites on
C-PC. UV-VIS spectrophotometry showed that the binding of hydrophobic (pro)vitamins
does not affect the protein colour, while CD spectroscopy revealed that the binding of
chosen molecules does not significantly influence the secondary structure of C-PC.
Overall, this study demonstrated C-PC's significant potential in binding hydrophobic
(pro)vitamins, while further research is required to test if these ligands could improve CPC
stability.
PB  - Serbian Biochemical Society
C3  - Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
T1  - Examination of C-phycocyanin interactions with selected vitamins
SP  - 106
EP  - 106
UR  - https://hdl.handle.net/21.15107/rcub_cer_6665
ER  - 

@conference{
author = "Ivanov, Aleksandar and Veličković, Luka and Jovanović, Zorana and Gligorijević, Nikola and Minić, Simeon and Nikolić, Milan",
year = "2023",
abstract = "C-phycocyanin (C-PC) is a photosynthetic protein from Arthrospira platensis
(cyanobacteria). Due to its intense blue colour, which is very rare in nature, C-PC has
industrial applications as a food colourant as a substitute for synthetic food colourants.
Disadvantages of C-PC as a food colourant are its poor stability at high temperatures
(during thermal treatment of the food) and its sensibility to change pH value. The binding
of food-derived small molecules, such as vitamins, could stabilize the structure of C-PC at
high temperatures and wide pH ranges. In this study, we characterized the binding of
selected vitamins to C-PC, purified from the commercial powder of Arthrospira platensis.
We used hydrophilic vitamins (B1, B2, B7, B9, B12), lipophilic vitamins (A, D3) and
provitamin (β-carotene). Fluorescent spectroscopy showed a decrease in fluorescence of CPC
i n t he p resence o f v itamin A, v itamin D3 a nd β -carotene (lipophilic molecules)
compared to the control. In contrast, the fluorescence of C-PC in the presence of
hydrophilic vitamins showed minimal change. The protein fluorescence quenching
approach demonstrated hydrophobic (pro)vitamins binding affinities ranging from 0.02 to
5.9 x 105 M-1, with the ability of hydrophobic (pro)vitamins to bind at the different sites on
C-PC. UV-VIS spectrophotometry showed that the binding of hydrophobic (pro)vitamins
does not affect the protein colour, while CD spectroscopy revealed that the binding of
chosen molecules does not significantly influence the secondary structure of C-PC.
Overall, this study demonstrated C-PC's significant potential in binding hydrophobic
(pro)vitamins, while further research is required to test if these ligands could improve CPC
stability.",
publisher = "Serbian Biochemical Society",
journal = "Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia",
title = "Examination of C-phycocyanin interactions with selected vitamins",
pages = "106-106",
url = "https://hdl.handle.net/21.15107/rcub_cer_6665"
}

Ivanov, A., Veličković, L., Jovanović, Z., Gligorijević, N., Minić, S.,& Nikolić, M.. (2023). Examination of C-phycocyanin interactions with selected vitamins. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
Serbian Biochemical Society., 106-106.
https://hdl.handle.net/21.15107/rcub_cer_6665

Ivanov A, Veličković L, Jovanović Z, Gligorijević N, Minić S, Nikolić M. Examination of C-phycocyanin interactions with selected vitamins. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia. 2023;:106-106.
https://hdl.handle.net/21.15107/rcub_cer_6665 .

Ivanov, Aleksandar, Veličković, Luka, Jovanović, Zorana, Gligorijević, Nikola, Minić, Simeon, Nikolić, Milan, "Examination of C-phycocyanin interactions with selected vitamins" in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia (2023):106-106,
https://hdl.handle.net/21.15107/rcub_cer_6665 .

TY  - CONF
AU  - Radović, Jelena
AU  - Popović, Dragana
AU  - Ćurčić, Tatjana
AU  - Nikolić, Milan
AU  - Minić, Simeon
AU  - Gligorijević, Nikola
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6666
AB  - This study aimed to improve chitosan polymer's capabilities to absorb mercury by
immobilising phycobiliproteins (PBPs) onto the surface of chitosan beads (chitosan–
PBPs). Phycobiliproteins, light-harvesting proteins from algae and cyanobacteria, have
several industrially essential applications. These proteins can bind heavy metals with high
affinities. Protein extracts obtained from both Arthrospira platensis, with C-phycocyanin
as the dominant phycobiliprotein and Neoporphyra haitanensis, with R-phycocyanin and
R-phycoerythrin as the dominant PBPs, were covalently immobilised onto chitosan beads.
Binding analysis showed that, on average, 54 μg of PBPs were immobilised per bead.
Immobilised proteins were still in their native state, with no visible colour change after
immobilisation. Chitosan–PBPs and chitosan alone were tested for mercury adsorption at
pH 4 and pH 7 by atomic absorption spectroscopy. The tested concentration range of
mercury was from 1 to 70 ppm. Affinity, calculated using Henry's binding isotherm, of
chitosan–PBPs for mercury was higher at both pH values than chitosan alone.
Furthermore, chitosan–PBPs beads were able to absorb significantly more mercury than
chitosan alone. These results show that the covalent immobilisation of PBPs onto chitosan
improves its mercury adsorption characteristics and creates a more efficient eco-friendly
adsorbent for removing mercury ions in the tested concentration range.
PB  - Serbian Biochemical Society
C3  - Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
T1  - Applying immobilised phycobiliproteins onto chitosan for efficient mercury removal
SP  - 116
EP  - 116
UR  - https://hdl.handle.net/21.15107/rcub_cer_6666
ER  - 

@conference{
author = "Radović, Jelena and Popović, Dragana and Ćurčić, Tatjana and Nikolić, Milan and Minić, Simeon and Gligorijević, Nikola",
year = "2023",
abstract = "This study aimed to improve chitosan polymer's capabilities to absorb mercury by
immobilising phycobiliproteins (PBPs) onto the surface of chitosan beads (chitosan–
PBPs). Phycobiliproteins, light-harvesting proteins from algae and cyanobacteria, have
several industrially essential applications. These proteins can bind heavy metals with high
affinities. Protein extracts obtained from both Arthrospira platensis, with C-phycocyanin
as the dominant phycobiliprotein and Neoporphyra haitanensis, with R-phycocyanin and
R-phycoerythrin as the dominant PBPs, were covalently immobilised onto chitosan beads.
Binding analysis showed that, on average, 54 μg of PBPs were immobilised per bead.
Immobilised proteins were still in their native state, with no visible colour change after
immobilisation. Chitosan–PBPs and chitosan alone were tested for mercury adsorption at
pH 4 and pH 7 by atomic absorption spectroscopy. The tested concentration range of
mercury was from 1 to 70 ppm. Affinity, calculated using Henry's binding isotherm, of
chitosan–PBPs for mercury was higher at both pH values than chitosan alone.
Furthermore, chitosan–PBPs beads were able to absorb significantly more mercury than
chitosan alone. These results show that the covalent immobilisation of PBPs onto chitosan
improves its mercury adsorption characteristics and creates a more efficient eco-friendly
adsorbent for removing mercury ions in the tested concentration range.",
publisher = "Serbian Biochemical Society",
journal = "Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia",
title = "Applying immobilised phycobiliproteins onto chitosan for efficient mercury removal",
pages = "116-116",
url = "https://hdl.handle.net/21.15107/rcub_cer_6666"
}

Radović, J., Popović, D., Ćurčić, T., Nikolić, M., Minić, S.,& Gligorijević, N.. (2023). Applying immobilised phycobiliproteins onto chitosan for efficient mercury removal. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
Serbian Biochemical Society., 116-116.
https://hdl.handle.net/21.15107/rcub_cer_6666

Radović J, Popović D, Ćurčić T, Nikolić M, Minić S, Gligorijević N. Applying immobilised phycobiliproteins onto chitosan for efficient mercury removal. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia. 2023;:116-116.
https://hdl.handle.net/21.15107/rcub_cer_6666 .

Radović, Jelena, Popović, Dragana, Ćurčić, Tatjana, Nikolić, Milan, Minić, Simeon, Gligorijević, Nikola, "Applying immobilised phycobiliproteins onto chitosan for efficient mercury removal" in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia (2023):116-116,
https://hdl.handle.net/21.15107/rcub_cer_6666 .

TY  - CONF
AU  - Aleksić, Ljubodrag
AU  - Veličković, Luka
AU  - Gligorijević, Nikola
AU  - Šunderić, Miloš
AU  - Takić, Marija
AU  - Nikolić, Milan
AU  - Minić, Simeon
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6667
AB  - Cultured meat requires less land and water and is less polluting, but still costly. The critical
challenge in cultivated meat science is identifying and developing bovine serum albumin
alternatives as the key component in cell media. Phycobiliproteins (PBPs) from micro- and
macroalgae are promising candidates for albumin replacement due to their high abundance
and well-known excellent antioxidative and metal-binding activities of covalently attached
tetrapyrrole chromophores. Considering the importance of fatty acids (FA) binding by
albumin for cell cultivation, the additional prerequisites for developing PBPs as albumin
replacement components is their validation for the ability to bind FA. This study aims to
examine the ability of C-phycocyanin (C-PC), the major PBP of microalgae Arthrospira
platensis, to bind seven fatty acids (stearic, palmitic, oleic, elaidic, linoleic, linolenic and
docosahexaenoic acid). For this purpose, we employed various optical spectroscopy
techniques (fluorescence, CD, and VIS absorption spectroscopy). The protein fluorescence
quenching approach demonstrated FA binding affinities ranging from 0.42 to 2.4 x 105
M−1, with the ability of FA to bind at different sites on C-PC. Fatty acid binding induces
substantial changes in the VIS absorption spectra of C-PC, indicating the FA are attached
in the vicinity of C-PC chromophores. On the other hand, CD spectroscopy did not show
significant effects of FA binding on C-PC secondary structure content. Overall, this study
revealed C-PC's significant potential in binding FA, the critical prerequisite to replacing
albumin for developing animal-free cell media for meat cultivation.
PB  - Serbian Biochemical Society
C3  - Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
T1  - Examining fatty acid interactions with Arthrospira platensis-derived C-phycocyanin
SP  - 121
EP  - 121
UR  - https://hdl.handle.net/21.15107/rcub_cer_6667
ER  - 

@conference{
author = "Aleksić, Ljubodrag and Veličković, Luka and Gligorijević, Nikola and Šunderić, Miloš and Takić, Marija and Nikolić, Milan and Minić, Simeon",
year = "2023",
abstract = "Cultured meat requires less land and water and is less polluting, but still costly. The critical
challenge in cultivated meat science is identifying and developing bovine serum albumin
alternatives as the key component in cell media. Phycobiliproteins (PBPs) from micro- and
macroalgae are promising candidates for albumin replacement due to their high abundance
and well-known excellent antioxidative and metal-binding activities of covalently attached
tetrapyrrole chromophores. Considering the importance of fatty acids (FA) binding by
albumin for cell cultivation, the additional prerequisites for developing PBPs as albumin
replacement components is their validation for the ability to bind FA. This study aims to
examine the ability of C-phycocyanin (C-PC), the major PBP of microalgae Arthrospira
platensis, to bind seven fatty acids (stearic, palmitic, oleic, elaidic, linoleic, linolenic and
docosahexaenoic acid). For this purpose, we employed various optical spectroscopy
techniques (fluorescence, CD, and VIS absorption spectroscopy). The protein fluorescence
quenching approach demonstrated FA binding affinities ranging from 0.42 to 2.4 x 105
M−1, with the ability of FA to bind at different sites on C-PC. Fatty acid binding induces
substantial changes in the VIS absorption spectra of C-PC, indicating the FA are attached
in the vicinity of C-PC chromophores. On the other hand, CD spectroscopy did not show
significant effects of FA binding on C-PC secondary structure content. Overall, this study
revealed C-PC's significant potential in binding FA, the critical prerequisite to replacing
albumin for developing animal-free cell media for meat cultivation.",
publisher = "Serbian Biochemical Society",
journal = "Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia",
title = "Examining fatty acid interactions with Arthrospira platensis-derived C-phycocyanin",
pages = "121-121",
url = "https://hdl.handle.net/21.15107/rcub_cer_6667"
}

Aleksić, L., Veličković, L., Gligorijević, N., Šunderić, M., Takić, M., Nikolić, M.,& Minić, S.. (2023). Examining fatty acid interactions with Arthrospira platensis-derived C-phycocyanin. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
Serbian Biochemical Society., 121-121.
https://hdl.handle.net/21.15107/rcub_cer_6667

Aleksić L, Veličković L, Gligorijević N, Šunderić M, Takić M, Nikolić M, Minić S. Examining fatty acid interactions with Arthrospira platensis-derived C-phycocyanin. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia. 2023;:121-121.
https://hdl.handle.net/21.15107/rcub_cer_6667 .

Aleksić, Ljubodrag, Veličković, Luka, Gligorijević, Nikola, Šunderić, Miloš, Takić, Marija, Nikolić, Milan, Minić, Simeon, "Examining fatty acid interactions with Arthrospira platensis-derived C-phycocyanin" in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia (2023):121-121,
https://hdl.handle.net/21.15107/rcub_cer_6667 .

TY  - CONF
AU  - Jovanović, Zorana
AU  - Annighöfer, Burkhard
AU  - Dudzinski, Daniel
AU  - Veličković, Luka
AU  - Gligorijević, Nikola
AU  - Nikolić, Milan
AU  - Brûlet, Annie
AU  - Assifaoui, Ali
AU  - Combet, Sophie
AU  - Minić, Simeon
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6669
AB  - The color of food products is an important aspect in food industry, and its preservation
remains a big challenge. We aim to preserve the natural blue dye of C-phycocyanin (C-PC)
phycobiliprotein from Spirulina microalgae. For this purpose, we incorporated C-PC in
combined starch and β-lactoglobulin (BLG) hydrogels by using a high-pressure (HP)
process. Indeed, in thermal treatment, the color derived from C-PC is entirely lost. We
characterized the obtained HP gels by both rheology and small-angle X-ray scattering
(SAXS). Various formulations of binary (BLG/C-PC) and ternary (starch/BLG/C-PC)
systems were tested under HP up to 4,500 bar. A good preservation of the C-PC pigment
was established by mixing BLG and starch with C-PC at pH 7, with concentrations of 180,
5, and 10 mg/mL, respectively. Identical component concentrations were maintained in the
binary systems. Structure of gels was characterized by SAXS providing insight of C-PC
interactions with BLG and starch after HP process which leads to the formation of solid
gels with larger mesh compared to two-component systems. This results in enhanced
mechanical properties, which were determined by amplitude and frequency sweep
measurements using a rheometer with applied plane/plane geometry. Therefore, adding
starch, even at small concentration, significantly improves gel visual appearance and
mechanical properties. Our study reveals that preservation through HP treatment is more
effective than high temperature treatment, as visually observed through the sustained color
integrity of C-PC blue dye.
PB  - Serbian Biochemical Society
C3  - Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
T1  - Combined hydrogels of starch and β-lactoglobulin as matrices for the preservation of C-phycocyanin
SP  - 150
EP  - 150
UR  - https://hdl.handle.net/21.15107/rcub_cer_6669
ER  - 

@conference{
author = "Jovanović, Zorana and Annighöfer, Burkhard and Dudzinski, Daniel and Veličković, Luka and Gligorijević, Nikola and Nikolić, Milan and Brûlet, Annie and Assifaoui, Ali and Combet, Sophie and Minić, Simeon",
year = "2023",
abstract = "The color of food products is an important aspect in food industry, and its preservation
remains a big challenge. We aim to preserve the natural blue dye of C-phycocyanin (C-PC)
phycobiliprotein from Spirulina microalgae. For this purpose, we incorporated C-PC in
combined starch and β-lactoglobulin (BLG) hydrogels by using a high-pressure (HP)
process. Indeed, in thermal treatment, the color derived from C-PC is entirely lost. We
characterized the obtained HP gels by both rheology and small-angle X-ray scattering
(SAXS). Various formulations of binary (BLG/C-PC) and ternary (starch/BLG/C-PC)
systems were tested under HP up to 4,500 bar. A good preservation of the C-PC pigment
was established by mixing BLG and starch with C-PC at pH 7, with concentrations of 180,
5, and 10 mg/mL, respectively. Identical component concentrations were maintained in the
binary systems. Structure of gels was characterized by SAXS providing insight of C-PC
interactions with BLG and starch after HP process which leads to the formation of solid
gels with larger mesh compared to two-component systems. This results in enhanced
mechanical properties, which were determined by amplitude and frequency sweep
measurements using a rheometer with applied plane/plane geometry. Therefore, adding
starch, even at small concentration, significantly improves gel visual appearance and
mechanical properties. Our study reveals that preservation through HP treatment is more
effective than high temperature treatment, as visually observed through the sustained color
integrity of C-PC blue dye.",
publisher = "Serbian Biochemical Society",
journal = "Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia",
title = "Combined hydrogels of starch and β-lactoglobulin as matrices for the preservation of C-phycocyanin",
pages = "150-150",
url = "https://hdl.handle.net/21.15107/rcub_cer_6669"
}

Jovanović, Z., Annighöfer, B., Dudzinski, D., Veličković, L., Gligorijević, N., Nikolić, M., Brûlet, A., Assifaoui, A., Combet, S.,& Minić, S.. (2023). Combined hydrogels of starch and β-lactoglobulin as matrices for the preservation of C-phycocyanin. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
Serbian Biochemical Society., 150-150.
https://hdl.handle.net/21.15107/rcub_cer_6669

Jovanović Z, Annighöfer B, Dudzinski D, Veličković L, Gligorijević N, Nikolić M, Brûlet A, Assifaoui A, Combet S, Minić S. Combined hydrogels of starch and β-lactoglobulin as matrices for the preservation of C-phycocyanin. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia. 2023;:150-150.
https://hdl.handle.net/21.15107/rcub_cer_6669 .

Jovanović, Zorana, Annighöfer, Burkhard, Dudzinski, Daniel, Veličković, Luka, Gligorijević, Nikola, Nikolić, Milan, Brûlet, Annie, Assifaoui, Ali, Combet, Sophie, Minić, Simeon, "Combined hydrogels of starch and β-lactoglobulin as matrices for the preservation of C-phycocyanin" in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia (2023):150-150,
https://hdl.handle.net/21.15107/rcub_cer_6669 .

TY  - CONF
AU  - Veličković, Luka
AU  - Sibinčić, Nikolina
AU  - Stojadinović, Marija
AU  - Gligorijević, Nikola
AU  - Nikolić, Milan
AU  - Srdić, Tatjana
AU  - Minić, Simeon
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6671
AB  - This study investigates the impact of Porphyra sp. extracts on HT29 cell line growth and
viability at reduced serum conditions. The concentration-dependent effects of
phycobiliproteins (PBPs) on cell proliferation were examined over various time intervals.
Lower concentrations of PBPs (20 μg/mL) demonstrated an increase in HT29 cell viability
after 48 hours and 5 days of cultivation at reduced serum concentration (final serum
concentration was in the range from 5 to 8%). This suggests a potential positive influence
on cell proliferation, likely due to their antioxidant properties. Conversely, higher
concentrations of PBPs exhibited inhibitory effects on cell growth, possibly due to
cytotoxicity at elevated levels. Remarkably, when HT29 cells were cultured solely in algal
extract without fetal calf serum (FCS), complete growth inhibition was observed after 72
hours. This finding underscores the insufficient nutrient and growth factor provision of
PBPs alone for sustaining cell viability. Morphological differences observed in cells
cultured with 70 μg/mL of PBPs indicated potential alterations in cellular morphology.
Notably, 70 μg/mL of PBPs in RPMI medium with 5% FCS displayed growth inhibition
compared to the control (5% FCS). Furthermore, we assessed HT29 cell adaptability to
changes in FCS concentration and PBP supplementation. Cells incubated under varying
FCS and PBP conditions were subcultured into RPMI medium with lower FCS
concentration and PBPs from Porphyra. The viability of cells following subculturing
indicated sustained adaptability to reduced FCS levels. Overall, this study provides
valuable insights into the concentration-dependent effects of PBPs from Porphyra extracts
on HT29 cell growth and viability. The findings underscore the potential benefits of PBPs
at lower concentrations for cell proliferation at reduced serum conditions and reveal the
adaptability of HT29 cells to changing culture conditions.
PB  - Serbian Biochemical Society
C3  - Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
T1  - Exploring if Porphyra sp. extract functions as serum substitute in HT29 cell culture
SP  - 82
EP  - 82
UR  - https://hdl.handle.net/21.15107/rcub_cer_6671
ER  - 

@conference{
author = "Veličković, Luka and Sibinčić, Nikolina and Stojadinović, Marija and Gligorijević, Nikola and Nikolić, Milan and Srdić, Tatjana and Minić, Simeon",
year = "2023",
abstract = "This study investigates the impact of Porphyra sp. extracts on HT29 cell line growth and
viability at reduced serum conditions. The concentration-dependent effects of
phycobiliproteins (PBPs) on cell proliferation were examined over various time intervals.
Lower concentrations of PBPs (20 μg/mL) demonstrated an increase in HT29 cell viability
after 48 hours and 5 days of cultivation at reduced serum concentration (final serum
concentration was in the range from 5 to 8%). This suggests a potential positive influence
on cell proliferation, likely due to their antioxidant properties. Conversely, higher
concentrations of PBPs exhibited inhibitory effects on cell growth, possibly due to
cytotoxicity at elevated levels. Remarkably, when HT29 cells were cultured solely in algal
extract without fetal calf serum (FCS), complete growth inhibition was observed after 72
hours. This finding underscores the insufficient nutrient and growth factor provision of
PBPs alone for sustaining cell viability. Morphological differences observed in cells
cultured with 70 μg/mL of PBPs indicated potential alterations in cellular morphology.
Notably, 70 μg/mL of PBPs in RPMI medium with 5% FCS displayed growth inhibition
compared to the control (5% FCS). Furthermore, we assessed HT29 cell adaptability to
changes in FCS concentration and PBP supplementation. Cells incubated under varying
FCS and PBP conditions were subcultured into RPMI medium with lower FCS
concentration and PBPs from Porphyra. The viability of cells following subculturing
indicated sustained adaptability to reduced FCS levels. Overall, this study provides
valuable insights into the concentration-dependent effects of PBPs from Porphyra extracts
on HT29 cell growth and viability. The findings underscore the potential benefits of PBPs
at lower concentrations for cell proliferation at reduced serum conditions and reveal the
adaptability of HT29 cells to changing culture conditions.",
publisher = "Serbian Biochemical Society",
journal = "Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia",
title = "Exploring if Porphyra sp. extract functions as serum substitute in HT29 cell culture",
pages = "82-82",
url = "https://hdl.handle.net/21.15107/rcub_cer_6671"
}

Veličković, L., Sibinčić, N., Stojadinović, M., Gligorijević, N., Nikolić, M., Srdić, T.,& Minić, S.. (2023). Exploring if Porphyra sp. extract functions as serum substitute in HT29 cell culture. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
Serbian Biochemical Society., 82-82.
https://hdl.handle.net/21.15107/rcub_cer_6671

Veličković L, Sibinčić N, Stojadinović M, Gligorijević N, Nikolić M, Srdić T, Minić S. Exploring if Porphyra sp. extract functions as serum substitute in HT29 cell culture. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia. 2023;:82-82.
https://hdl.handle.net/21.15107/rcub_cer_6671 .

Veličković, Luka, Sibinčić, Nikolina, Stojadinović, Marija, Gligorijević, Nikola, Nikolić, Milan, Srdić, Tatjana, Minić, Simeon, "Exploring if Porphyra sp. extract functions as serum substitute in HT29 cell culture" in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia (2023):82-82,
https://hdl.handle.net/21.15107/rcub_cer_6671 .

TY  - CONF
AU  - Gligorijević, Nikola
AU  - Stanić-Vučinić, Dragana
AU  - Mutić, Tamara
AU  - Lujić, Tamara
AU  - Ćirković Veličković, Tanja
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6524
AB  - Microplastic represents one of the major types of pollutants in modern era. Over several years of research in the field of microplastic, there are still many unknown gaps, including the effects and mechanisms of action of these particles on human health. Studies in this field conducted experiments on cells and human tissues or animals like rats and mice. While these studies suggest the toxic effects of microplastic, it is not clear if concentrations used for exposure are relevant for humans. Also, most of the studies used spherical polystyrene, which does not reflect well the diversity of microplastic particles found in nature. Another gap is lack of studies describing direct interactions of microplastics and proteins. While it is generally known that proteins form corona around microplastic particles, affinity studies and consequences on protein structure are usually missing. The aim of this work was to analyze interaction of a major egg white protein and allergen, ovalbumin to several to microplastic particles, including polystyrene (PS) of 120 and 500 μm in size and polyethylene terephthalate (PET) of 120 μm in size. Binding affinity was determined at both acidic, pH 3 and neutral, pH 7 conditions, at the room temperature, by measuring bulk ovalbumin concentration in supernatants at the equilibrium time. Several binding models, including Langmuir,
Freundlich, Redlich–Peterson and Guggenheim-Anderson-de Boer (GAB), were used to determine binding parameters. The formation of soft and hard corona was analyzed according to the published protocol [1]. Structural analysis was performed using near and far-UV CD spectrometry.
Obtained results showed that ovalbumin binds to both PS and PET. All binding models indicated that ovalbumin binds with higher affinity to tested microplastics on pH 3, compared to pH 7, with the highest affinity being calculated for PS 120 μm. Further analysis showed that ovalbumin forms both soft and hard corona onto the surface of all three microplastics. Structural alterations of ovalbumin as a consequence of its interaction with microplastic was shown to be both pH and
microplastic type dependent. Also, more pronounced effect on its tertiary structure was observed, compared to secondary. At pH3, tertiary structure of bulk ovalbumin becomes destabilized, especially in the presence of PET 120 μm and PS 500 μm, while at pH 7, structural stabilization is observed, especially in the presence of PS 120 μm. Considering that the microplastic was discovered in eggs [2], obtained results suggest that direct interactions of native ovalbumin with microplastic particles could have influence on its structure and thus affect its techno-functional properties.
PB  - Belgrade : Serbian Chemical Society
C3  - Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023
T1  - Binding and corona formation of ovalbumin to polystyrene and polyethylene terephthalate microplastics under neutral and acidic conditions
SP  - 137
EP  - 137
UR  - https://hdl.handle.net/21.15107/rcub_cer_6524
ER  - 

@conference{
author = "Gligorijević, Nikola and Stanić-Vučinić, Dragana and Mutić, Tamara and Lujić, Tamara and Ćirković Veličković, Tanja",
year = "2023",
abstract = "Microplastic represents one of the major types of pollutants in modern era. Over several years of research in the field of microplastic, there are still many unknown gaps, including the effects and mechanisms of action of these particles on human health. Studies in this field conducted experiments on cells and human tissues or animals like rats and mice. While these studies suggest the toxic effects of microplastic, it is not clear if concentrations used for exposure are relevant for humans. Also, most of the studies used spherical polystyrene, which does not reflect well the diversity of microplastic particles found in nature. Another gap is lack of studies describing direct interactions of microplastics and proteins. While it is generally known that proteins form corona around microplastic particles, affinity studies and consequences on protein structure are usually missing. The aim of this work was to analyze interaction of a major egg white protein and allergen, ovalbumin to several to microplastic particles, including polystyrene (PS) of 120 and 500 μm in size and polyethylene terephthalate (PET) of 120 μm in size. Binding affinity was determined at both acidic, pH 3 and neutral, pH 7 conditions, at the room temperature, by measuring bulk ovalbumin concentration in supernatants at the equilibrium time. Several binding models, including Langmuir,
Freundlich, Redlich–Peterson and Guggenheim-Anderson-de Boer (GAB), were used to determine binding parameters. The formation of soft and hard corona was analyzed according to the published protocol [1]. Structural analysis was performed using near and far-UV CD spectrometry.
Obtained results showed that ovalbumin binds to both PS and PET. All binding models indicated that ovalbumin binds with higher affinity to tested microplastics on pH 3, compared to pH 7, with the highest affinity being calculated for PS 120 μm. Further analysis showed that ovalbumin forms both soft and hard corona onto the surface of all three microplastics. Structural alterations of ovalbumin as a consequence of its interaction with microplastic was shown to be both pH and
microplastic type dependent. Also, more pronounced effect on its tertiary structure was observed, compared to secondary. At pH3, tertiary structure of bulk ovalbumin becomes destabilized, especially in the presence of PET 120 μm and PS 500 μm, while at pH 7, structural stabilization is observed, especially in the presence of PS 120 μm. Considering that the microplastic was discovered in eggs [2], obtained results suggest that direct interactions of native ovalbumin with microplastic particles could have influence on its structure and thus affect its techno-functional properties.",
publisher = "Belgrade : Serbian Chemical Society",
journal = "Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023",
title = "Binding and corona formation of ovalbumin to polystyrene and polyethylene terephthalate microplastics under neutral and acidic conditions",
pages = "137-137",
url = "https://hdl.handle.net/21.15107/rcub_cer_6524"
}

Gligorijević, N., Stanić-Vučinić, D., Mutić, T., Lujić, T.,& Ćirković Veličković, T.. (2023). Binding and corona formation of ovalbumin to polystyrene and polyethylene terephthalate microplastics under neutral and acidic conditions. in Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023
Belgrade : Serbian Chemical Society., 137-137.
https://hdl.handle.net/21.15107/rcub_cer_6524

Gligorijević N, Stanić-Vučinić D, Mutić T, Lujić T, Ćirković Veličković T. Binding and corona formation of ovalbumin to polystyrene and polyethylene terephthalate microplastics under neutral and acidic conditions. in Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023. 2023;:137-137.
https://hdl.handle.net/21.15107/rcub_cer_6524 .

Gligorijević, Nikola, Stanić-Vučinić, Dragana, Mutić, Tamara, Lujić, Tamara, Ćirković Veličković, Tanja, "Binding and corona formation of ovalbumin to polystyrene and polyethylene terephthalate microplastics under neutral and acidic conditions" in Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023 (2023):137-137,
https://hdl.handle.net/21.15107/rcub_cer_6524 .

TY  - CONF
AU  - Lujić, Tamara
AU  - Gligorijević, Nikola
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković Veličković, Tanja
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6525
AB  - Ovalbumin (OVA) is the most abundant protein in chicken egg white. It is one of the major allergens in eggs. Micro- and nanoplatic particles (MNPs) are a widespread contaminant and have been found in food and water. It is still unclear how MNPs might affect human health. However, due to their large surface area they have been found to bind various biopolymers,
including proteins. These biopolymers can be bound more strongly or loosely, and are referred to as hard and soft corona, respectfully [1]. MPs have been found in eggs, in the size range of 50-100 μm [2]. It is shown that these particles can interact with proteins and induce structural changes, but there is still not enough information on this topic [3]. These structural changes could lead to a decreased digestibility in the gastrointestinal tract, which could increase the immune
response to known allergens. The aim of this study was to determine whether there are structural changes present in the OVA after incubation with two types of MPs – 120 μm polyethylene terephthalate (PET) and 120 μm polystyrene (PS) and whether they could influence
digestion of OVA with gastrointestinal enzymes. 20 mg of MPs were incubated with 1.3 mg/mL ovalbumin for 4 h at room temperature in a 20 mM phosphate buffer at pH 7. Bulk ovalbumin was separated from the MPs by centrifugation and by filtration through a 0.22 μm PVDF filter. Soft corona was obtained by washing the MPs with water, and the MPs were later removed as described with bulk ovalbumin. Formation of amyloids was monitored with a Thioflavin T (ThT) assay at room temperature and after thermal treatment, and additional structural analysis was performed by circular dichroism (CD) spectrometry in the far-UV region. Thermal stability was also determined by spectrofluorimetry. Digestion with two proteases (pepsin and trypsin) was performed to determine whether there is a change in the gastrointestinal digestibility of OVA.
Results from the ThT assay show that at room temperature there is no significant difference between the fluorescence emission obtained for all samples, with bulk OVA from both MPs showing a slight decrease. However, there is an increase of fluorescence after thermal treatment in all OVA samples, where OVA from the soft corona emits significantly less fluorescence
than control and bulk samples for both types of MPs. Additionally, soft coronas have been shown to have more β-sheet content than other samples, which is more pronounced for OVA incubated with PET. For the heated samples there is a sharp change from α-helix to β-sheets in all the samples, but it is the most dramatic in the soft coronas. This could impose rigidity to the tertiary structure, which would explain why the ThT molecule does not bind as strongly. Despite differences in both the secondary and tertiary structure, the thermal stability is almost the same in all samples. Digestion of the samples shows that the soft corona incubated with PS tends to be more resistant to trypsin than other samples after 2 min, but it is not significant. For digestion with pepsin there is no difference between the samples. In conjunction with the previous results, which indicates a structural stabilisation of the soft corona at pH 7, it is not surprising that there is an increased resistance to trypsin, compared to pepsin which is a gastric enzyme and for which digestion is performed at an acidic pH. In conclusion, there is a structural change present in samples upon contact with MPs, particularly in the soft corona, of which the most pronounced is a decrease of α-helix content and increase in β-sheet content as determined by far-UV CD.
This leads to a structural stabilization which could further impact the digestibility of the OVA protein and impact its allergenicity. However, this must be confirmed with further experiments.
PB  - Belgrade : Serbian Chemical Society
C3  - Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023
T1  - Investigation of structural changes in ovalbumin induced by two types of MPs and its impact on protein digestibility
SP  - 153
EP  - 153
UR  - https://hdl.handle.net/21.15107/rcub_cer_6525
ER  - 

@conference{
author = "Lujić, Tamara and Gligorijević, Nikola and Stanić-Vučinić, Dragana and Ćirković Veličković, Tanja",
year = "2023",
abstract = "Ovalbumin (OVA) is the most abundant protein in chicken egg white. It is one of the major allergens in eggs. Micro- and nanoplatic particles (MNPs) are a widespread contaminant and have been found in food and water. It is still unclear how MNPs might affect human health. However, due to their large surface area they have been found to bind various biopolymers,
including proteins. These biopolymers can be bound more strongly or loosely, and are referred to as hard and soft corona, respectfully [1]. MPs have been found in eggs, in the size range of 50-100 μm [2]. It is shown that these particles can interact with proteins and induce structural changes, but there is still not enough information on this topic [3]. These structural changes could lead to a decreased digestibility in the gastrointestinal tract, which could increase the immune
response to known allergens. The aim of this study was to determine whether there are structural changes present in the OVA after incubation with two types of MPs – 120 μm polyethylene terephthalate (PET) and 120 μm polystyrene (PS) and whether they could influence
digestion of OVA with gastrointestinal enzymes. 20 mg of MPs were incubated with 1.3 mg/mL ovalbumin for 4 h at room temperature in a 20 mM phosphate buffer at pH 7. Bulk ovalbumin was separated from the MPs by centrifugation and by filtration through a 0.22 μm PVDF filter. Soft corona was obtained by washing the MPs with water, and the MPs were later removed as described with bulk ovalbumin. Formation of amyloids was monitored with a Thioflavin T (ThT) assay at room temperature and after thermal treatment, and additional structural analysis was performed by circular dichroism (CD) spectrometry in the far-UV region. Thermal stability was also determined by spectrofluorimetry. Digestion with two proteases (pepsin and trypsin) was performed to determine whether there is a change in the gastrointestinal digestibility of OVA.
Results from the ThT assay show that at room temperature there is no significant difference between the fluorescence emission obtained for all samples, with bulk OVA from both MPs showing a slight decrease. However, there is an increase of fluorescence after thermal treatment in all OVA samples, where OVA from the soft corona emits significantly less fluorescence
than control and bulk samples for both types of MPs. Additionally, soft coronas have been shown to have more β-sheet content than other samples, which is more pronounced for OVA incubated with PET. For the heated samples there is a sharp change from α-helix to β-sheets in all the samples, but it is the most dramatic in the soft coronas. This could impose rigidity to the tertiary structure, which would explain why the ThT molecule does not bind as strongly. Despite differences in both the secondary and tertiary structure, the thermal stability is almost the same in all samples. Digestion of the samples shows that the soft corona incubated with PS tends to be more resistant to trypsin than other samples after 2 min, but it is not significant. For digestion with pepsin there is no difference between the samples. In conjunction with the previous results, which indicates a structural stabilisation of the soft corona at pH 7, it is not surprising that there is an increased resistance to trypsin, compared to pepsin which is a gastric enzyme and for which digestion is performed at an acidic pH. In conclusion, there is a structural change present in samples upon contact with MPs, particularly in the soft corona, of which the most pronounced is a decrease of α-helix content and increase in β-sheet content as determined by far-UV CD.
This leads to a structural stabilization which could further impact the digestibility of the OVA protein and impact its allergenicity. However, this must be confirmed with further experiments.",
publisher = "Belgrade : Serbian Chemical Society",
journal = "Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023",
title = "Investigation of structural changes in ovalbumin induced by two types of MPs and its impact on protein digestibility",
pages = "153-153",
url = "https://hdl.handle.net/21.15107/rcub_cer_6525"
}

Lujić, T., Gligorijević, N., Stanić-Vučinić, D.,& Ćirković Veličković, T.. (2023). Investigation of structural changes in ovalbumin induced by two types of MPs and its impact on protein digestibility. in Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023
Belgrade : Serbian Chemical Society., 153-153.
https://hdl.handle.net/21.15107/rcub_cer_6525

Lujić T, Gligorijević N, Stanić-Vučinić D, Ćirković Veličković T. Investigation of structural changes in ovalbumin induced by two types of MPs and its impact on protein digestibility. in Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023. 2023;:153-153.
https://hdl.handle.net/21.15107/rcub_cer_6525 .

Lujić, Tamara, Gligorijević, Nikola, Stanić-Vučinić, Dragana, Ćirković Veličković, Tanja, "Investigation of structural changes in ovalbumin induced by two types of MPs and its impact on protein digestibility" in Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023 (2023):153-153,
https://hdl.handle.net/21.15107/rcub_cer_6525 .

TY  - CONF
AU  - Četić, Danilo
AU  - Miljuš, Goran
AU  - Dobrijević, Zorana
AU  - Gligorijević, Nikola
AU  - Vilotić, Aleksandra
AU  - Nedić, Olgica
AU  - Penezić, Ana
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6663
AB  - Human transferrin (Tf) is a bilobal 76 kDa iron-binding glycoprotein present in human
serum. Each lobe has the ability to bind one ferric ion (Fe3+) and a single synergistic
bicarbonate anion. The main role of Tf is to transport Fe3+ ions through the circulation to
cells, via interaction with transferrin receptor (TFR) on the cell surface. Previously
described methods for Tf isolation and purification are either very time-consuming or
provide Tf of lower final purity. Here we describe a fast and simple FPLC method for the
isolation and purification of Tf from human serum. Serum samples were prepared by
precipitation, while protein purification was performed on FPLC system, using an anionexchange
column. Several different buffers at the same pH were tested. Tf purified by this
method was analyzed by Western blot, followed by immunodetection, as well as with
silver staining after SDS PAGE. Its functionality was tested with respect to iron-binding
capacity (ferozzine method) and its ability to interact with TFR by immunofluorescent
staining. The conformation of purified Tf was analyzed by recording intrinsic fluorescent
emmision spectra originating from Trp residues. The method itself is highly reproducible
(intra- and interday), easy to perform (only two steps) and fast (under an hour), yielding
98% to 99% pure Tf with all buffers. Purified Tf was shown to have retained its ironbinding
capacity, as well as the ability to interact with TFR. Purified Tf also retained its
native three-dimensional structure. Described method for the isolation and purification of
Tf is fast, simple and highly reproducible, yielding a functional Tf of high purity in its
native state while offering the flexibility of using different buffer systems. All of these
features make this protocol a method of choice for the isolation and purification of Tf on a
semi-preparative scale.
PB  - Serbian Biochemical Society
C3  - Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
T1  - Simple two-step semi-preparative isolation and purification of transferrin from human serum
SP  - 66
EP  - 66
UR  - https://hdl.handle.net/21.15107/rcub_cer_6663
ER  - 

@conference{
author = "Četić, Danilo and Miljuš, Goran and Dobrijević, Zorana and Gligorijević, Nikola and Vilotić, Aleksandra and Nedić, Olgica and Penezić, Ana",
year = "2023",
abstract = "Human transferrin (Tf) is a bilobal 76 kDa iron-binding glycoprotein present in human
serum. Each lobe has the ability to bind one ferric ion (Fe3+) and a single synergistic
bicarbonate anion. The main role of Tf is to transport Fe3+ ions through the circulation to
cells, via interaction with transferrin receptor (TFR) on the cell surface. Previously
described methods for Tf isolation and purification are either very time-consuming or
provide Tf of lower final purity. Here we describe a fast and simple FPLC method for the
isolation and purification of Tf from human serum. Serum samples were prepared by
precipitation, while protein purification was performed on FPLC system, using an anionexchange
column. Several different buffers at the same pH were tested. Tf purified by this
method was analyzed by Western blot, followed by immunodetection, as well as with
silver staining after SDS PAGE. Its functionality was tested with respect to iron-binding
capacity (ferozzine method) and its ability to interact with TFR by immunofluorescent
staining. The conformation of purified Tf was analyzed by recording intrinsic fluorescent
emmision spectra originating from Trp residues. The method itself is highly reproducible
(intra- and interday), easy to perform (only two steps) and fast (under an hour), yielding
98% to 99% pure Tf with all buffers. Purified Tf was shown to have retained its ironbinding
capacity, as well as the ability to interact with TFR. Purified Tf also retained its
native three-dimensional structure. Described method for the isolation and purification of
Tf is fast, simple and highly reproducible, yielding a functional Tf of high purity in its
native state while offering the flexibility of using different buffer systems. All of these
features make this protocol a method of choice for the isolation and purification of Tf on a
semi-preparative scale.",
publisher = "Serbian Biochemical Society",
journal = "Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia",
title = "Simple two-step semi-preparative isolation and purification of transferrin from human serum",
pages = "66-66",
url = "https://hdl.handle.net/21.15107/rcub_cer_6663"
}

Četić, D., Miljuš, G., Dobrijević, Z., Gligorijević, N., Vilotić, A., Nedić, O.,& Penezić, A.. (2023). Simple two-step semi-preparative isolation and purification of transferrin from human serum. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
Serbian Biochemical Society., 66-66.
https://hdl.handle.net/21.15107/rcub_cer_6663

Četić D, Miljuš G, Dobrijević Z, Gligorijević N, Vilotić A, Nedić O, Penezić A. Simple two-step semi-preparative isolation and purification of transferrin from human serum. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia. 2023;:66-66.
https://hdl.handle.net/21.15107/rcub_cer_6663 .

Četić, Danilo, Miljuš, Goran, Dobrijević, Zorana, Gligorijević, Nikola, Vilotić, Aleksandra, Nedić, Olgica, Penezić, Ana, "Simple two-step semi-preparative isolation and purification of transferrin from human serum" in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia (2023):66-66,
https://hdl.handle.net/21.15107/rcub_cer_6663 .

TY  - CONF
AU  - Miljuš, Goran
AU  - Penezić, Ana
AU  - Noe, Dragana
AU  - Dobrijević, Zorana
AU  - Baralić, Marko
AU  - Robajac, Dragana
AU  - Šunderić, Miloš
AU  - Gligorijević, Nikola
AU  - Dimitrijević, Ivan
AU  - Barišić, Goran
AU  - Nedić, Olgica
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6664
AB  - Human serum tansferrin (hsTf) is a major circulatory protein crucial for the
transport/metabolism of Fe3+ ions. By sequestering and delivering ferric ions to target
tissues/cells hsTf maintains redox homeostasis. Oxidative stress (OS), one of the hallmarks
of (patho)physiological conditions, alters protein structure and function. The main role of
hsTf hinges on specific interaction with cellular Tf receptor (TfR) while other interactions
contribute to diverse functions. The aim of this study was to profile interacting partners of
hsTf in the samples of serum coming from patients diagnosed with a wide range of
pathological conditions with underlying OS status. Anti-hsTf pull-down samples were
analysed using mass spectrometry. Data went through analysis by appropriate
bioinformatic tools. Results reveal differences in expression of hsTf interacting proteins in
sample groups of patients suffering from kidney insufficiency subjected to dialysis
treatment (peritoneal-PD or hemo-HD) also with patients with gestational diabetes
compared to respective healthy sample groups. Colorectal cancer stage T3 versus T2 stage
shows an inverse distribution of expression profiles in comparison to healthy samples.
Most prominent differences are seen in hsTf interacting partners involved in the
complement and coagulation cascades and cholesterol metabolic pathways, suggesting a
multifaceted role of hsTf in these processes throughout the course of the disease.
PB  - Serbian Biochemical Society
C3  - Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
T1  - Proteomic profiling of anti-transferrin pull-down in patients with underlying oxidative stress
SP  - 69
EP  - 69
UR  - https://hdl.handle.net/21.15107/rcub_cer_6664
ER  - 

@conference{
author = "Miljuš, Goran and Penezić, Ana and Noe, Dragana and Dobrijević, Zorana and Baralić, Marko and Robajac, Dragana and Šunderić, Miloš and Gligorijević, Nikola and Dimitrijević, Ivan and Barišić, Goran and Nedić, Olgica",
year = "2023",
abstract = "Human serum tansferrin (hsTf) is a major circulatory protein crucial for the
transport/metabolism of Fe3+ ions. By sequestering and delivering ferric ions to target
tissues/cells hsTf maintains redox homeostasis. Oxidative stress (OS), one of the hallmarks
of (patho)physiological conditions, alters protein structure and function. The main role of
hsTf hinges on specific interaction with cellular Tf receptor (TfR) while other interactions
contribute to diverse functions. The aim of this study was to profile interacting partners of
hsTf in the samples of serum coming from patients diagnosed with a wide range of
pathological conditions with underlying OS status. Anti-hsTf pull-down samples were
analysed using mass spectrometry. Data went through analysis by appropriate
bioinformatic tools. Results reveal differences in expression of hsTf interacting proteins in
sample groups of patients suffering from kidney insufficiency subjected to dialysis
treatment (peritoneal-PD or hemo-HD) also with patients with gestational diabetes
compared to respective healthy sample groups. Colorectal cancer stage T3 versus T2 stage
shows an inverse distribution of expression profiles in comparison to healthy samples.
Most prominent differences are seen in hsTf interacting partners involved in the
complement and coagulation cascades and cholesterol metabolic pathways, suggesting a
multifaceted role of hsTf in these processes throughout the course of the disease.",
publisher = "Serbian Biochemical Society",
journal = "Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia",
title = "Proteomic profiling of anti-transferrin pull-down in patients with underlying oxidative stress",
pages = "69-69",
url = "https://hdl.handle.net/21.15107/rcub_cer_6664"
}

Miljuš, G., Penezić, A., Noe, D., Dobrijević, Z., Baralić, M., Robajac, D., Šunderić, M., Gligorijević, N., Dimitrijević, I., Barišić, G.,& Nedić, O.. (2023). Proteomic profiling of anti-transferrin pull-down in patients with underlying oxidative stress. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
Serbian Biochemical Society., 69-69.
https://hdl.handle.net/21.15107/rcub_cer_6664

Miljuš G, Penezić A, Noe D, Dobrijević Z, Baralić M, Robajac D, Šunderić M, Gligorijević N, Dimitrijević I, Barišić G, Nedić O. Proteomic profiling of anti-transferrin pull-down in patients with underlying oxidative stress. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia. 2023;:69-69.
https://hdl.handle.net/21.15107/rcub_cer_6664 .

Miljuš, Goran, Penezić, Ana, Noe, Dragana, Dobrijević, Zorana, Baralić, Marko, Robajac, Dragana, Šunderić, Miloš, Gligorijević, Nikola, Dimitrijević, Ivan, Barišić, Goran, Nedić, Olgica, "Proteomic profiling of anti-transferrin pull-down in patients with underlying oxidative stress" in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia (2023):69-69,
https://hdl.handle.net/21.15107/rcub_cer_6664 .

TY  - CONF
AU  - Radomirović, Mirjana
AU  - Gligorijević, Nikola
AU  - Stanić-Vučinić, Dragana
AU  - Rajković, Andreja
AU  - Ćirković Veličković, Tanja
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6668
AB  - Tropomyosin has been recognized as one of the most common allergens among shellfish
allergens. Sensitive and specific quantification of traces of allergens present in food
samples is of critical importance for people with food allergies. This study thus aimed to
develop a highly sensitive immuno-PCR method for detecting crustacean tropomyosin in
foods. Method couples conventional sandwich ELISA assay with real-time PCR
amplification of marker DNA. Monoclonal mouse anti-tropomyosin antibody was used as
a capture antibody, while polyclonal rabbit anti-tropomyosin antibody served as a
detection antibody in sandwich ELISA. A double-stranded amino-DNA molecule of 77
base pairs was covalently conjugated to a secondary goat anti-rabbit antibody and
subsequently amplified and quantified by real-time PCR. Tropomyosin was quantified
using highly purified natural shrimp tropomyosin as standard. The sensitivity of immuno-
PCR for quantification of tropomyosin was increased by up to 20-fold compared to
ELISA, demonstrating a ccuracy a s l ow a s 1 9.8 p g/mL. Recovery of tropomyosin in
vegetable soup as a food matrix was in the 87.7–115.6% range, with relative standard
deviations in the 5–24.5% range. Tropomyosin was also quantified in the commercially
available food products. Developed immuno-PCR technique thus shows the potential to be
a method of choice for specific and ultrasensitive detection of tropomyosin in food
samples, with the final aim of reducing risks of accidental food contamination.
PB  - Serbian Biochemical Society
C3  - Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
T1  - Immuno-PCR for crustacean tropomyosin quantification
SP  - 130
EP  - 130
UR  - https://hdl.handle.net/21.15107/rcub_cer_6668
ER  - 

@conference{
author = "Radomirović, Mirjana and Gligorijević, Nikola and Stanić-Vučinić, Dragana and Rajković, Andreja and Ćirković Veličković, Tanja",
year = "2023",
abstract = "Tropomyosin has been recognized as one of the most common allergens among shellfish
allergens. Sensitive and specific quantification of traces of allergens present in food
samples is of critical importance for people with food allergies. This study thus aimed to
develop a highly sensitive immuno-PCR method for detecting crustacean tropomyosin in
foods. Method couples conventional sandwich ELISA assay with real-time PCR
amplification of marker DNA. Monoclonal mouse anti-tropomyosin antibody was used as
a capture antibody, while polyclonal rabbit anti-tropomyosin antibody served as a
detection antibody in sandwich ELISA. A double-stranded amino-DNA molecule of 77
base pairs was covalently conjugated to a secondary goat anti-rabbit antibody and
subsequently amplified and quantified by real-time PCR. Tropomyosin was quantified
using highly purified natural shrimp tropomyosin as standard. The sensitivity of immuno-
PCR for quantification of tropomyosin was increased by up to 20-fold compared to
ELISA, demonstrating a ccuracy a s l ow a s 1 9.8 p g/mL. Recovery of tropomyosin in
vegetable soup as a food matrix was in the 87.7–115.6% range, with relative standard
deviations in the 5–24.5% range. Tropomyosin was also quantified in the commercially
available food products. Developed immuno-PCR technique thus shows the potential to be
a method of choice for specific and ultrasensitive detection of tropomyosin in food
samples, with the final aim of reducing risks of accidental food contamination.",
publisher = "Serbian Biochemical Society",
journal = "Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia",
title = "Immuno-PCR for crustacean tropomyosin quantification",
pages = "130-130",
url = "https://hdl.handle.net/21.15107/rcub_cer_6668"
}

Radomirović, M., Gligorijević, N., Stanić-Vučinić, D., Rajković, A.,& Ćirković Veličković, T.. (2023). Immuno-PCR for crustacean tropomyosin quantification. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
Serbian Biochemical Society., 130-130.
https://hdl.handle.net/21.15107/rcub_cer_6668

Radomirović M, Gligorijević N, Stanić-Vučinić D, Rajković A, Ćirković Veličković T. Immuno-PCR for crustacean tropomyosin quantification. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia. 2023;:130-130.
https://hdl.handle.net/21.15107/rcub_cer_6668 .

Radomirović, Mirjana, Gligorijević, Nikola, Stanić-Vučinić, Dragana, Rajković, Andreja, Ćirković Veličković, Tanja, "Immuno-PCR for crustacean tropomyosin quantification" in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia (2023):130-130,
https://hdl.handle.net/21.15107/rcub_cer_6668 .

TY  - CONF
AU  - Lujić, Tamara
AU  - Gligorijević, Nikola
AU  - Stanić-Vučinić, Dragana
AU  - Bićanin, Maša
AU  - Ćirković Veličković, Tanja
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6651
AB  - Mircoplastics (MPs) are an abundant contaminant in the environment with ingestion being the most common way of exposure for humans. Binding of protein to MPs is proposed to be multilayered with the formation of a soft and hard corona. It has been proven that MPs interact with enzymes present in the digestive system and impact their activity. The aim of this study is to investigate the impact of MPs on the activity of trypsin in simulated intestinal fluid (SIF). For this purpose, two sizes of polypropylene (large – 180-500 μm, small – 63-180 μm) and one size of polyethylene terephthalate (<80 μm) have been studied. Activity in bulk and soft corona was determined in SIF at 405 nm with N-α- Benzoyl-DL-arginine 4-nitroanilide hydrochloride after different times of incubation. Activity in hard corona was determined after 1 h of incubation with the MPs. Although specific activity in the control decreases through time, there is a tendency for all MPs to preserve activity in bulk and soft corona trypsin after 4 h of incubation. Trypsin remains active in the hard corona, with the activity being an order of magnitude lower than in the control, possibly due to significant changes in structure.
PB  - Serbian Biochemical Society
C3  - Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
T1  - Impact of MPs on trypsin activity in simulated intestinal fluid
SP  - 145
EP  - 145
UR  - https://hdl.handle.net/21.15107/rcub_cer_6651
ER  - 

@conference{
author = "Lujić, Tamara and Gligorijević, Nikola and Stanić-Vučinić, Dragana and Bićanin, Maša and Ćirković Veličković, Tanja",
year = "2023",
abstract = "Mircoplastics (MPs) are an abundant contaminant in the environment with ingestion being the most common way of exposure for humans. Binding of protein to MPs is proposed to be multilayered with the formation of a soft and hard corona. It has been proven that MPs interact with enzymes present in the digestive system and impact their activity. The aim of this study is to investigate the impact of MPs on the activity of trypsin in simulated intestinal fluid (SIF). For this purpose, two sizes of polypropylene (large – 180-500 μm, small – 63-180 μm) and one size of polyethylene terephthalate (<80 μm) have been studied. Activity in bulk and soft corona was determined in SIF at 405 nm with N-α- Benzoyl-DL-arginine 4-nitroanilide hydrochloride after different times of incubation. Activity in hard corona was determined after 1 h of incubation with the MPs. Although specific activity in the control decreases through time, there is a tendency for all MPs to preserve activity in bulk and soft corona trypsin after 4 h of incubation. Trypsin remains active in the hard corona, with the activity being an order of magnitude lower than in the control, possibly due to significant changes in structure.",
publisher = "Serbian Biochemical Society",
journal = "Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia",
title = "Impact of MPs on trypsin activity in simulated intestinal fluid",
pages = "145-145",
url = "https://hdl.handle.net/21.15107/rcub_cer_6651"
}

Lujić, T., Gligorijević, N., Stanić-Vučinić, D., Bićanin, M.,& Ćirković Veličković, T.. (2023). Impact of MPs on trypsin activity in simulated intestinal fluid. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
Serbian Biochemical Society., 145-145.
https://hdl.handle.net/21.15107/rcub_cer_6651

Lujić T, Gligorijević N, Stanić-Vučinić D, Bićanin M, Ćirković Veličković T. Impact of MPs on trypsin activity in simulated intestinal fluid. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia. 2023;:145-145.
https://hdl.handle.net/21.15107/rcub_cer_6651 .

Lujić, Tamara, Gligorijević, Nikola, Stanić-Vučinić, Dragana, Bićanin, Maša, Ćirković Veličković, Tanja, "Impact of MPs on trypsin activity in simulated intestinal fluid" in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia (2023):145-145,
https://hdl.handle.net/21.15107/rcub_cer_6651 .

TY  - JOUR
AU  - Radomirović, Mirjana
AU  - Gligorijević, Nikola
AU  - Stanić-Vučinić, Dragana
AU  - Rajković, Andreja
AU  - Ćirković Veličković, Tanja
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6782
AB  - Tropomyosin is the major and predominant allergen among shellfish. This study developed
an ultrasensitive immuno-PCR method for the quantification of crustacean tropomyosin in foods.
The method couples sandwich ELISA with the real-time PCR (rtPCR) amplification of marker DNAs.
Monoclonal anti-TPM antibody was the capture antibody, polyclonal rabbit anti-shrimp tropomyosin
antibody was the detection antibody, while natural shrimp tropomyosin served as the standard. A
double-stranded amino-DNA was covalently conjugated to a secondary anti-rabbit antibody and
subsequently amplified and quantified via rtPCR. The quantification sensitivity of immuno-PCR
was 20-fold higher than analogous ELISA, with LOQ 19.8 pg/mL. The developed immuno-PCR
method is highly specific for the detection of crustacean tropomyosin and is highly precise in a
broad concentration range. Tropomyosin recovery in the spiked vegetable soup was 87.7–115.6%.
Crustacean tropomyosin was also quantified in commercial food products. The reported immuno-
PCR assay is the most sensitive method for the quantification of crustacean tropomyosin and is the
first immuno-PCR-based assay for the quantification of food allergen and food protein in general.
The described method could be easily adapted for the specific and ultrasensitive immuno-PCR-based
detection of traces of any food allergen that is currently being quantified with ELISA, which is of
critical importance for people with food allergies.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - Ultrasensitive Quantification of Crustacean Tropomyosin by Immuno-PCR
VL  - 24
IS  - 20
SP  - 15410
DO  - 10.3390/ijms242015410
ER  - 

@article{
author = "Radomirović, Mirjana and Gligorijević, Nikola and Stanić-Vučinić, Dragana and Rajković, Andreja and Ćirković Veličković, Tanja",
year = "2023",
abstract = "Tropomyosin is the major and predominant allergen among shellfish. This study developed
an ultrasensitive immuno-PCR method for the quantification of crustacean tropomyosin in foods.
The method couples sandwich ELISA with the real-time PCR (rtPCR) amplification of marker DNAs.
Monoclonal anti-TPM antibody was the capture antibody, polyclonal rabbit anti-shrimp tropomyosin
antibody was the detection antibody, while natural shrimp tropomyosin served as the standard. A
double-stranded amino-DNA was covalently conjugated to a secondary anti-rabbit antibody and
subsequently amplified and quantified via rtPCR. The quantification sensitivity of immuno-PCR
was 20-fold higher than analogous ELISA, with LOQ 19.8 pg/mL. The developed immuno-PCR
method is highly specific for the detection of crustacean tropomyosin and is highly precise in a
broad concentration range. Tropomyosin recovery in the spiked vegetable soup was 87.7–115.6%.
Crustacean tropomyosin was also quantified in commercial food products. The reported immuno-
PCR assay is the most sensitive method for the quantification of crustacean tropomyosin and is the
first immuno-PCR-based assay for the quantification of food allergen and food protein in general.
The described method could be easily adapted for the specific and ultrasensitive immuno-PCR-based
detection of traces of any food allergen that is currently being quantified with ELISA, which is of
critical importance for people with food allergies.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "Ultrasensitive Quantification of Crustacean Tropomyosin by Immuno-PCR",
volume = "24",
number = "20",
pages = "15410",
doi = "10.3390/ijms242015410"
}

Radomirović, M., Gligorijević, N., Stanić-Vučinić, D., Rajković, A.,& Ćirković Veličković, T.. (2023). Ultrasensitive Quantification of Crustacean Tropomyosin by Immuno-PCR. in International Journal of Molecular Sciences
MDPI., 24(20), 15410.
https://doi.org/10.3390/ijms242015410

Radomirović M, Gligorijević N, Stanić-Vučinić D, Rajković A, Ćirković Veličković T. Ultrasensitive Quantification of Crustacean Tropomyosin by Immuno-PCR. in International Journal of Molecular Sciences. 2023;24(20):15410.
doi:10.3390/ijms242015410 .

Radomirović, Mirjana, Gligorijević, Nikola, Stanić-Vučinić, Dragana, Rajković, Andreja, Ćirković Veličković, Tanja, "Ultrasensitive Quantification of Crustacean Tropomyosin by Immuno-PCR" in International Journal of Molecular Sciences, 24, no. 20 (2023):15410,
https://doi.org/10.3390/ijms242015410 . .

TY  - CONF
AU  - Gligorijević, Nikola
AU  - Lujić, Tamara
AU  - Mutić, Tamara
AU  - Vasović, Tamara
AU  - de Guzman, Maria Krishna
AU  - Aćimović, Jelena
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković Veličković, Tanja
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6898
AB  - Ovalbumin (OVA), a main protein of egg white, has characteristic structural fold of a
serpin-family of proteins, propensity to fibril formation and stability to digestion.
Microplastics (MPs) contaminating our food can interact with food proteins in the food
matrix and during digestion. In this study adsorption of OVA to polystyrene (PS) (110 μm
and 260 μm), polyethylene terephthalate (PET) (140 μm) MPs were investigated in acidic
(pH 3) and neutral (pH 7) conditions. Formations of corona on MPs were investigated
using isolated OVA and egg white protein extract comparatively. OVA adsorption depends
on MPs size, polymer chemistry and pH, being highest in acidic pH and higher for PS.
Adsorption of OVA to PS and PET reaches dynamic equilibrium after 4h resulting in
disruption of tertiary structure and formation of hard and soft corona around MPs. Shorter
fragments of OVA populate hard corona, while soft corona exclusively consist of full
length OVA, albeit in its non-native conformation. The conformational changes resemble
those induced by heat treatment with re-arrangement of α-β secondary structures.
Structural changes are striking for the OVA in corona around MPs. Soft corona OVA
preserves thermal and proteolytic stability, but loses ability to form fibrils upon heating.
OVA is abundantly present in corona around MPs also in the presence of other egg white
proteins. MPs contaminating food may bind and change structure and functional properties
of main egg white protein.
PB  - Kragujevac, Srbija : Prirodno-matematički fakultet, Univerzitet u Kragujevcu /  Kragujevac, Serbia : Faculty of Science, University of Kragujevac
C3  - VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike,” Zbornik apstrakata, 2. jun 2023. godine, Kragujevac / VI Symposium of a Serbian proteomic society: „Discussion and Application of  New Methods of Proteomics,“ Book of abstratcs,
T1  - Biocorona formation of hen egg white proteins onto the surface of polystyrene and polyethylene terephthalate
SP  - OP10
EP  - OP10
UR  - https://hdl.handle.net/21.15107/rcub_cer_6898
ER  - 

@conference{
author = "Gligorijević, Nikola and Lujić, Tamara and Mutić, Tamara and Vasović, Tamara and de Guzman, Maria Krishna and Aćimović, Jelena and Stanić-Vučinić, Dragana and Ćirković Veličković, Tanja",
year = "2023",
abstract = "Ovalbumin (OVA), a main protein of egg white, has characteristic structural fold of a
serpin-family of proteins, propensity to fibril formation and stability to digestion.
Microplastics (MPs) contaminating our food can interact with food proteins in the food
matrix and during digestion. In this study adsorption of OVA to polystyrene (PS) (110 μm
and 260 μm), polyethylene terephthalate (PET) (140 μm) MPs were investigated in acidic
(pH 3) and neutral (pH 7) conditions. Formations of corona on MPs were investigated
using isolated OVA and egg white protein extract comparatively. OVA adsorption depends
on MPs size, polymer chemistry and pH, being highest in acidic pH and higher for PS.
Adsorption of OVA to PS and PET reaches dynamic equilibrium after 4h resulting in
disruption of tertiary structure and formation of hard and soft corona around MPs. Shorter
fragments of OVA populate hard corona, while soft corona exclusively consist of full
length OVA, albeit in its non-native conformation. The conformational changes resemble
those induced by heat treatment with re-arrangement of α-β secondary structures.
Structural changes are striking for the OVA in corona around MPs. Soft corona OVA
preserves thermal and proteolytic stability, but loses ability to form fibrils upon heating.
OVA is abundantly present in corona around MPs also in the presence of other egg white
proteins. MPs contaminating food may bind and change structure and functional properties
of main egg white protein.",
publisher = "Kragujevac, Srbija : Prirodno-matematički fakultet, Univerzitet u Kragujevcu /  Kragujevac, Serbia : Faculty of Science, University of Kragujevac",
journal = "VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike,” Zbornik apstrakata, 2. jun 2023. godine, Kragujevac / VI Symposium of a Serbian proteomic society: „Discussion and Application of  New Methods of Proteomics,“ Book of abstratcs,",
title = "Biocorona formation of hen egg white proteins onto the surface of polystyrene and polyethylene terephthalate",
pages = "OP10-OP10",
url = "https://hdl.handle.net/21.15107/rcub_cer_6898"
}

Gligorijević, N., Lujić, T., Mutić, T., Vasović, T., de Guzman, M. K., Aćimović, J., Stanić-Vučinić, D.,& Ćirković Veličković, T.. (2023). Biocorona formation of hen egg white proteins onto the surface of polystyrene and polyethylene terephthalate. in VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike,” Zbornik apstrakata, 2. jun 2023. godine, Kragujevac / VI Symposium of a Serbian proteomic society: „Discussion and Application of  New Methods of Proteomics,“ Book of abstratcs,
Kragujevac, Srbija : Prirodno-matematički fakultet, Univerzitet u Kragujevcu /  Kragujevac, Serbia : Faculty of Science, University of Kragujevac., OP10-OP10.
https://hdl.handle.net/21.15107/rcub_cer_6898

Gligorijević N, Lujić T, Mutić T, Vasović T, de Guzman MK, Aćimović J, Stanić-Vučinić D, Ćirković Veličković T. Biocorona formation of hen egg white proteins onto the surface of polystyrene and polyethylene terephthalate. in VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike,” Zbornik apstrakata, 2. jun 2023. godine, Kragujevac / VI Symposium of a Serbian proteomic society: „Discussion and Application of  New Methods of Proteomics,“ Book of abstratcs,. 2023;:OP10-OP10.
https://hdl.handle.net/21.15107/rcub_cer_6898 .

Gligorijević, Nikola, Lujić, Tamara, Mutić, Tamara, Vasović, Tamara, de Guzman, Maria Krishna, Aćimović, Jelena, Stanić-Vučinić, Dragana, Ćirković Veličković, Tanja, "Biocorona formation of hen egg white proteins onto the surface of polystyrene and polyethylene terephthalate" in VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike,” Zbornik apstrakata, 2. jun 2023. godine, Kragujevac / VI Symposium of a Serbian proteomic society: „Discussion and Application of  New Methods of Proteomics,“ Book of abstratcs, (2023):OP10-OP10,
https://hdl.handle.net/21.15107/rcub_cer_6898 .

TY  - CONF
AU  - Lujić, Tamara
AU  - Gligorijević, Nikola
AU  - Jovanović, Vesna B.
AU  - Aćimović, Jelena
AU  - Mitić, Dragana
AU  - Vasović, Tamara
AU  - Stojadinović, Marija
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7473
AB  - Microplastics is abundant in the environment, food and beverages and get ingested by humans.
Its complex interplay with proteins lead to formation of corona. Tightly bound proteins represent
hard corona, while weaker binding partners are found in soft corona. Separation of hard and soft
corona of allergenic proteins of shrimps, eggs and cow’s milk, tropomyosin (TPM), ovalbumin
(OVA) and beta-lactoglobulin (BLG) and identification of binding partners by proteomics was
aim of our study.
Allergenic proteins were purified from egg white, shrimps and cow’s milk. Binding to
polyethylene terephthalate microplastics (PET) (70-100 m) was probed at pH 7 for purified
allergens and egg white proteins. After establishment of binding equilibrium, soft and hard
corona were separated and analyzed by SDS PAGE, followed by identification of bound
proteins by nanoLC-HRMS. Binding of all allergenic proteins was observed in both soft and
hard corona. Soft corona contains exclusively intact, full length OVA, TPM and BLG. Hard
corona is enriched for truncated OVA and oligomers of TPM. OVA fragments are partially or
fully enfolded and have higher level of exposed hydrophobic patches resulting in higher affinity
for PET microplastics. In comparison to OVA and TPM, hard corona of BLG is less abundant
under similar conditions. BLG is compact globular protein with lower level of exposed
hydrophobic patches in comparison to ovalbumin and tropomyosin. In hard corona, trace
amounts of contaminating alfa-lactalbumin become enriched. In the presence of egg white
protein extract OVA forms both SC and HC on microplastics, being the dominant protein of
hard corona (with ovotransferrin). Lysozyme and ovomucin are present only in hard corona.
Both proteins are known for their strong bioactivity and represent a small fraction of total egg
white proteins.
Our results show that allergenic proteins form hard corona on PET microplastics. Among egg
white proteins, minor proteins such as lysozyme and ovomucin become enriched. Denaturing
effect of strong binding to microplastics may change functional characteristics of allergens and
bioactive proteins of foods and should be further investigated in functional assays.
PB  - Italian Proteomics Association
C3  - XVII International Italian Proteomics Association Annual Meeting in partnership with the Hellenic Proteomics Society and Serbian Proteomics Association, "Proteomics and Metabolomics towards Global Health", November 29th-December 1st, 2023, Ospedale Isola Tiberina - Gemelli Isola,  Roma, Italy
T1  - Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics
SP  - 11
EP  - 11
UR  - https://hdl.handle.net/21.15107/rcub_cer_7473
ER  - 

@conference{
author = "Lujić, Tamara and Gligorijević, Nikola and Jovanović, Vesna B. and Aćimović, Jelena and Mitić, Dragana and Vasović, Tamara and Stojadinović, Marija and Stanić-Vučinić, Dragana and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "Microplastics is abundant in the environment, food and beverages and get ingested by humans.
Its complex interplay with proteins lead to formation of corona. Tightly bound proteins represent
hard corona, while weaker binding partners are found in soft corona. Separation of hard and soft
corona of allergenic proteins of shrimps, eggs and cow’s milk, tropomyosin (TPM), ovalbumin
(OVA) and beta-lactoglobulin (BLG) and identification of binding partners by proteomics was
aim of our study.
Allergenic proteins were purified from egg white, shrimps and cow’s milk. Binding to
polyethylene terephthalate microplastics (PET) (70-100 m) was probed at pH 7 for purified
allergens and egg white proteins. After establishment of binding equilibrium, soft and hard
corona were separated and analyzed by SDS PAGE, followed by identification of bound
proteins by nanoLC-HRMS. Binding of all allergenic proteins was observed in both soft and
hard corona. Soft corona contains exclusively intact, full length OVA, TPM and BLG. Hard
corona is enriched for truncated OVA and oligomers of TPM. OVA fragments are partially or
fully enfolded and have higher level of exposed hydrophobic patches resulting in higher affinity
for PET microplastics. In comparison to OVA and TPM, hard corona of BLG is less abundant
under similar conditions. BLG is compact globular protein with lower level of exposed
hydrophobic patches in comparison to ovalbumin and tropomyosin. In hard corona, trace
amounts of contaminating alfa-lactalbumin become enriched. In the presence of egg white
protein extract OVA forms both SC and HC on microplastics, being the dominant protein of
hard corona (with ovotransferrin). Lysozyme and ovomucin are present only in hard corona.
Both proteins are known for their strong bioactivity and represent a small fraction of total egg
white proteins.
Our results show that allergenic proteins form hard corona on PET microplastics. Among egg
white proteins, minor proteins such as lysozyme and ovomucin become enriched. Denaturing
effect of strong binding to microplastics may change functional characteristics of allergens and
bioactive proteins of foods and should be further investigated in functional assays.",
publisher = "Italian Proteomics Association",
journal = "XVII International Italian Proteomics Association Annual Meeting in partnership with the Hellenic Proteomics Society and Serbian Proteomics Association, "Proteomics and Metabolomics towards Global Health", November 29th-December 1st, 2023, Ospedale Isola Tiberina - Gemelli Isola,  Roma, Italy",
title = "Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics",
pages = "11-11",
url = "https://hdl.handle.net/21.15107/rcub_cer_7473"
}

Lujić, T., Gligorijević, N., Jovanović, V. B., Aćimović, J., Mitić, D., Vasović, T., Stojadinović, M., Stanić-Vučinić, D.,& Ćirković-Veličković, T.. (2023). Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics. in XVII International Italian Proteomics Association Annual Meeting in partnership with the Hellenic Proteomics Society and Serbian Proteomics Association, "Proteomics and Metabolomics towards Global Health", November 29th-December 1st, 2023, Ospedale Isola Tiberina - Gemelli Isola,  Roma, Italy
Italian Proteomics Association., 11-11.
https://hdl.handle.net/21.15107/rcub_cer_7473

Lujić T, Gligorijević N, Jovanović VB, Aćimović J, Mitić D, Vasović T, Stojadinović M, Stanić-Vučinić D, Ćirković-Veličković T. Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics. in XVII International Italian Proteomics Association Annual Meeting in partnership with the Hellenic Proteomics Society and Serbian Proteomics Association, "Proteomics and Metabolomics towards Global Health", November 29th-December 1st, 2023, Ospedale Isola Tiberina - Gemelli Isola,  Roma, Italy. 2023;:11-11.
https://hdl.handle.net/21.15107/rcub_cer_7473 .

Lujić, Tamara, Gligorijević, Nikola, Jovanović, Vesna B., Aćimović, Jelena, Mitić, Dragana, Vasović, Tamara, Stojadinović, Marija, Stanić-Vučinić, Dragana, Ćirković-Veličković, Tanja, "Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics" in XVII International Italian Proteomics Association Annual Meeting in partnership with the Hellenic Proteomics Society and Serbian Proteomics Association, "Proteomics and Metabolomics towards Global Health", November 29th-December 1st, 2023, Ospedale Isola Tiberina - Gemelli Isola,  Roma, Italy (2023):11-11,
https://hdl.handle.net/21.15107/rcub_cer_7473 .

TY  - JOUR
AU  - de Guzman, Maria Krishna
AU  - Stanić-Vučinić, Dragana
AU  - Gligorijević, Nikola
AU  - Wimmer, Lukas
AU  - Gasparyan, Manvel
AU  - Lujić, Tamara
AU  - Vasović, Tamara
AU  - Dailey, Lea Ann
AU  - Van Haute, Sam
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6466
AB  - Human ingestion of microplastics (MPs) is common and inevitable due to the widespread contamination of food items, but implications on the gastric digestion of food proteins are still unknown. In this study, the interactions between pepsin and polystyrene (PS) MPs were evaluated by investigating enzyme activity and conformation in a simulated human gastric environment in the presence or absence of PS MPs. The impact on food digestion was also assessed by monitoring the kinetics of protein hydrolysis through static in vitro gastric digestion of cow's milk contaminated with PS. The binding of pepsin to PS showed that the surface chemistry of MPs dictates binding affinity. The key contributor to pepsin adsorption seems to be π−π interactions between the aromatic residues and the PS phenyl rings. During quick exposure (10 min) of pepsin to increasing concentrations (222, 2219, 22188 particles/mL) of 10 μm PS (PS10) and 100 μm PS (PS100), total enzymatic activities were not affected remarkably. However, upon prolonged exposure at 1 and 2 h, preferential binding of pepsin to the small, low zeta-potential PS caused structural changes in the protein which led to a significant reduction of its activity. Digestion of cow's milk mixed with PS10 resulted in transient accumulation of larger peptides (10–35 kDa) and reduced bioavailability of short peptides (2–9 kDa) in the gastric phase. This, however, was only observed at extremely high PS10 concentration (0.3 mg/mL or 5.46E+05 particles/mL). The digestion of milk peptides, bound preferentially over pepsin within the hard corona on the PS10 surface, was delayed up to 15 min in comparison to bulk protein digestion. Intact caseins, otherwise rapidly digested, remained bound to PS10 in the hard corona for up to 15 min. This work presents valuable insights regarding the interaction of MPs, food proteins, and pepsin, and their dynamics during gastric digestion.
PB  - Elsevier Ltd.
T2  - Environmental Pollution
T1  - Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion
VL  - 335
SP  - 122282
DO  - 10.1016/j.envpol.2023.122282
ER  - 

@article{
author = "de Guzman, Maria Krishna and Stanić-Vučinić, Dragana and Gligorijević, Nikola and Wimmer, Lukas and Gasparyan, Manvel and Lujić, Tamara and Vasović, Tamara and Dailey, Lea Ann and Van Haute, Sam and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "Human ingestion of microplastics (MPs) is common and inevitable due to the widespread contamination of food items, but implications on the gastric digestion of food proteins are still unknown. In this study, the interactions between pepsin and polystyrene (PS) MPs were evaluated by investigating enzyme activity and conformation in a simulated human gastric environment in the presence or absence of PS MPs. The impact on food digestion was also assessed by monitoring the kinetics of protein hydrolysis through static in vitro gastric digestion of cow's milk contaminated with PS. The binding of pepsin to PS showed that the surface chemistry of MPs dictates binding affinity. The key contributor to pepsin adsorption seems to be π−π interactions between the aromatic residues and the PS phenyl rings. During quick exposure (10 min) of pepsin to increasing concentrations (222, 2219, 22188 particles/mL) of 10 μm PS (PS10) and 100 μm PS (PS100), total enzymatic activities were not affected remarkably. However, upon prolonged exposure at 1 and 2 h, preferential binding of pepsin to the small, low zeta-potential PS caused structural changes in the protein which led to a significant reduction of its activity. Digestion of cow's milk mixed with PS10 resulted in transient accumulation of larger peptides (10–35 kDa) and reduced bioavailability of short peptides (2–9 kDa) in the gastric phase. This, however, was only observed at extremely high PS10 concentration (0.3 mg/mL or 5.46E+05 particles/mL). The digestion of milk peptides, bound preferentially over pepsin within the hard corona on the PS10 surface, was delayed up to 15 min in comparison to bulk protein digestion. Intact caseins, otherwise rapidly digested, remained bound to PS10 in the hard corona for up to 15 min. This work presents valuable insights regarding the interaction of MPs, food proteins, and pepsin, and their dynamics during gastric digestion.",
publisher = "Elsevier Ltd.",
journal = "Environmental Pollution",
title = "Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion",
volume = "335",
pages = "122282",
doi = "10.1016/j.envpol.2023.122282"
}

de Guzman, M. K., Stanić-Vučinić, D., Gligorijević, N., Wimmer, L., Gasparyan, M., Lujić, T., Vasović, T., Dailey, L. A., Van Haute, S.,& Ćirković-Veličković, T.. (2023). Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion. in Environmental Pollution
Elsevier Ltd.., 335, 122282.
https://doi.org/10.1016/j.envpol.2023.122282

de Guzman MK, Stanić-Vučinić D, Gligorijević N, Wimmer L, Gasparyan M, Lujić T, Vasović T, Dailey LA, Van Haute S, Ćirković-Veličković T. Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion. in Environmental Pollution. 2023;335:122282.
doi:10.1016/j.envpol.2023.122282 .

de Guzman, Maria Krishna, Stanić-Vučinić, Dragana, Gligorijević, Nikola, Wimmer, Lukas, Gasparyan, Manvel, Lujić, Tamara, Vasović, Tamara, Dailey, Lea Ann, Van Haute, Sam, Ćirković-Veličković, Tanja, "Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion" in Environmental Pollution, 335 (2023):122282,
https://doi.org/10.1016/j.envpol.2023.122282 . .

TY  - JOUR
AU  - Baralić, Marko
AU  - Robajac, Dragana
AU  - Penezić, Ana
AU  - Brković, Voin
AU  - Gligorijević, Nikola
AU  - Bontić, Ana
AU  - Pavlović, Jelena
AU  - Nikolić, Jelena
AU  - Miljuš, Goran
AU  - Dobrijević, Zorana
AU  - Šunderić, Miloš
AU  - Pažitna, Lucija
AU  - Katrlík, Jaroslav
AU  - Nedić, Olgica
AU  - Laušević, Mirjana
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6445
AB  - In previous publications, we pointed out the importance of mannosylation of fibrinogen for the development of cardiovascular complications and fucosylation as a predictor of peritoneal membrane dysfunction in patients on peritoneal dialysis (PD). After a follow-up period of 30 months from the onset of the COVID-19 pandemic, we evaluated the significance of 1,25-dihydroxyvitamin D3 (calcitriol) therapy, primary disease, biochemical and hematologic analyzes, and previously performed glycan analysis by lectin-based microarray as predictors of mortality in this patient group. After univariate Cox regression analysis, diabetes mellitus (DM) and calcitriol therapy were found to be potential predictors of mortality. Additional multivariate Cox regression analysis confirmed that only DM was a predictor of mortality. Nevertheless, the use of calcitriol in therapy significantly reduced mortality in this patient group, as shown by the Kaplan–Meier survival curve. The presence of DM as a concomitant disease proved to be a strong predictor of fatal outcome in PD patients infected with SARS-CoV-2. This is the first study to indicate the importance and beneficial effect of calcitriol therapy on survival in PD patients with COVID-19 infection. In addition, this study points to the possibility that adverse thrombogenic events observed in PD patients during the pandemic may be caused by aberrant fibrinogen glycosylation.
PB  - Switzerland : Multidisciplinary Digital Publishing Institute (MDPI)
T2  - Nutrients
T1  - Significance of 1,25-Dihydroxyvitamin D3 on Overall Mortality in Peritoneal Dialysis Patients with COVID-19
VL  - 15
IS  - 9
SP  - 2050
DO  - 10.3390/nu15092050
ER  - 

@article{
author = "Baralić, Marko and Robajac, Dragana and Penezić, Ana and Brković, Voin and Gligorijević, Nikola and Bontić, Ana and Pavlović, Jelena and Nikolić, Jelena and Miljuš, Goran and Dobrijević, Zorana and Šunderić, Miloš and Pažitna, Lucija and Katrlík, Jaroslav and Nedić, Olgica and Laušević, Mirjana",
year = "2023",
abstract = "In previous publications, we pointed out the importance of mannosylation of fibrinogen for the development of cardiovascular complications and fucosylation as a predictor of peritoneal membrane dysfunction in patients on peritoneal dialysis (PD). After a follow-up period of 30 months from the onset of the COVID-19 pandemic, we evaluated the significance of 1,25-dihydroxyvitamin D3 (calcitriol) therapy, primary disease, biochemical and hematologic analyzes, and previously performed glycan analysis by lectin-based microarray as predictors of mortality in this patient group. After univariate Cox regression analysis, diabetes mellitus (DM) and calcitriol therapy were found to be potential predictors of mortality. Additional multivariate Cox regression analysis confirmed that only DM was a predictor of mortality. Nevertheless, the use of calcitriol in therapy significantly reduced mortality in this patient group, as shown by the Kaplan–Meier survival curve. The presence of DM as a concomitant disease proved to be a strong predictor of fatal outcome in PD patients infected with SARS-CoV-2. This is the first study to indicate the importance and beneficial effect of calcitriol therapy on survival in PD patients with COVID-19 infection. In addition, this study points to the possibility that adverse thrombogenic events observed in PD patients during the pandemic may be caused by aberrant fibrinogen glycosylation.",
publisher = "Switzerland : Multidisciplinary Digital Publishing Institute (MDPI)",
journal = "Nutrients",
title = "Significance of 1,25-Dihydroxyvitamin D3 on Overall Mortality in Peritoneal Dialysis Patients with COVID-19",
volume = "15",
number = "9",
pages = "2050",
doi = "10.3390/nu15092050"
}

Baralić, M., Robajac, D., Penezić, A., Brković, V., Gligorijević, N., Bontić, A., Pavlović, J., Nikolić, J., Miljuš, G., Dobrijević, Z., Šunderić, M., Pažitna, L., Katrlík, J., Nedić, O.,& Laušević, M.. (2023). Significance of 1,25-Dihydroxyvitamin D3 on Overall Mortality in Peritoneal Dialysis Patients with COVID-19. in Nutrients
Switzerland : Multidisciplinary Digital Publishing Institute (MDPI)., 15(9), 2050.
https://doi.org/10.3390/nu15092050

Baralić M, Robajac D, Penezić A, Brković V, Gligorijević N, Bontić A, Pavlović J, Nikolić J, Miljuš G, Dobrijević Z, Šunderić M, Pažitna L, Katrlík J, Nedić O, Laušević M. Significance of 1,25-Dihydroxyvitamin D3 on Overall Mortality in Peritoneal Dialysis Patients with COVID-19. in Nutrients. 2023;15(9):2050.
doi:10.3390/nu15092050 .

Baralić, Marko, Robajac, Dragana, Penezić, Ana, Brković, Voin, Gligorijević, Nikola, Bontić, Ana, Pavlović, Jelena, Nikolić, Jelena, Miljuš, Goran, Dobrijević, Zorana, Šunderić, Miloš, Pažitna, Lucija, Katrlík, Jaroslav, Nedić, Olgica, Laušević, Mirjana, "Significance of 1,25-Dihydroxyvitamin D3 on Overall Mortality in Peritoneal Dialysis Patients with COVID-19" in Nutrients, 15, no. 9 (2023):2050,
https://doi.org/10.3390/nu15092050 . .