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  - 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  - 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  - Radomirović, Mirjana
AU  - Gligorijević, Nikola
AU  - Minić, Simeon
AU  - Nikolić, Milan
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković Veličković, Tanja
PY  - 2022
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6945
AB  - Phycocyanobilin (PCB) is an open-chain tetrapyrrole chromophore of phycocyanin (PC), chromoprotein derived from the cyanobacterium Arthrospira platensis. Our group has previously demonstrated the potential of PCB to covalently modify free cysteine residue of proteins using food protein β-lactoglobulin as a model protein. Relying on the proven ability of PCB to be attached to sulfhydryl groups of proteins, we propose a new method for covalent attachment of PCB to potentially any protein. We used Traut’s reagent (TR, 2-iminothiolane) to introduce free sulfhydryl groups in the model protein, bovine serum albumin (BSA), by modifying its lysine residues. All tested molar ratios of TR per mole of protein successfully modified BSA. A higher degree of modification by TR induced more profound alterations of BSA structure, as evidenced by near-UV and far-UV circular dichroism spectroscopy. At the same time, minor changes in BSA oligomerization and aggregation profile occurred with increasing TR molar ratio. PCB was covalently attached to introduced sulfhydryl groups at pH 9 at 20–fold molar ratio of PCB per mole of protein. An increase in the molar ratio of TR per mole of BSA leads to amplification of fluorescent signal of PCB-modified BSA, most significantly observed starting from 50-fold and higher TR ratios. Using BSA as a model protein, a 50-fold molar excess of TR seems to be the optimal choice for balancing a satisfactory signal amplification level and the negative effect on protein structure. BSA covalently modified with PCB has higher antioxidative activity than free BSA. The proposed method thus serves as a proof of concept for labeling virtually any protein with PCB as means of either functionalization through covalent attachment of bioactive PCB or obtaining fluorescent probes for application in fluorescence-based techniques.
PB  - Wiley
PB  - Federation of European Biochemical Societies
C3  - The Biochemistry Global Summit, 9th-14th July, 2022. In: FEBS Open Bio
T1  - Covalent modification of bovine serum albumin with phycocyanobilin using Traut's reagent
VL  - 12
IS  - Suppl. 1
SP  - 302
EP  - 303
UR  - https://hdl.handle.net/21.15107/rcub_cer_6945
ER  - 

@conference{
author = "Radomirović, Mirjana and Gligorijević, Nikola and Minić, Simeon and Nikolić, Milan and Stanić-Vučinić, Dragana and Ćirković Veličković, Tanja",
year = "2022",
abstract = "Phycocyanobilin (PCB) is an open-chain tetrapyrrole chromophore of phycocyanin (PC), chromoprotein derived from the cyanobacterium Arthrospira platensis. Our group has previously demonstrated the potential of PCB to covalently modify free cysteine residue of proteins using food protein β-lactoglobulin as a model protein. Relying on the proven ability of PCB to be attached to sulfhydryl groups of proteins, we propose a new method for covalent attachment of PCB to potentially any protein. We used Traut’s reagent (TR, 2-iminothiolane) to introduce free sulfhydryl groups in the model protein, bovine serum albumin (BSA), by modifying its lysine residues. All tested molar ratios of TR per mole of protein successfully modified BSA. A higher degree of modification by TR induced more profound alterations of BSA structure, as evidenced by near-UV and far-UV circular dichroism spectroscopy. At the same time, minor changes in BSA oligomerization and aggregation profile occurred with increasing TR molar ratio. PCB was covalently attached to introduced sulfhydryl groups at pH 9 at 20–fold molar ratio of PCB per mole of protein. An increase in the molar ratio of TR per mole of BSA leads to amplification of fluorescent signal of PCB-modified BSA, most significantly observed starting from 50-fold and higher TR ratios. Using BSA as a model protein, a 50-fold molar excess of TR seems to be the optimal choice for balancing a satisfactory signal amplification level and the negative effect on protein structure. BSA covalently modified with PCB has higher antioxidative activity than free BSA. The proposed method thus serves as a proof of concept for labeling virtually any protein with PCB as means of either functionalization through covalent attachment of bioactive PCB or obtaining fluorescent probes for application in fluorescence-based techniques.",
publisher = "Wiley, Federation of European Biochemical Societies",
journal = "The Biochemistry Global Summit, 9th-14th July, 2022. In: FEBS Open Bio",
title = "Covalent modification of bovine serum albumin with phycocyanobilin using Traut's reagent",
volume = "12",
number = "Suppl. 1",
pages = "302-303",
url = "https://hdl.handle.net/21.15107/rcub_cer_6945"
}

Radomirović, M., Gligorijević, N., Minić, S., Nikolić, M., Stanić-Vučinić, D.,& Ćirković Veličković, T.. (2022). Covalent modification of bovine serum albumin with phycocyanobilin using Traut's reagent. in The Biochemistry Global Summit, 9th-14th July, 2022. In: FEBS Open Bio
Wiley., 12(Suppl. 1), 302-303.
https://hdl.handle.net/21.15107/rcub_cer_6945

Radomirović M, Gligorijević N, Minić S, Nikolić M, Stanić-Vučinić D, Ćirković Veličković T. Covalent modification of bovine serum albumin with phycocyanobilin using Traut's reagent. in The Biochemistry Global Summit, 9th-14th July, 2022. In: FEBS Open Bio. 2022;12(Suppl. 1):302-303.
https://hdl.handle.net/21.15107/rcub_cer_6945 .

Radomirović, Mirjana, Gligorijević, Nikola, Minić, Simeon, Nikolić, Milan, Stanić-Vučinić, Dragana, Ćirković Veličković, Tanja, "Covalent modification of bovine serum albumin with phycocyanobilin using Traut's reagent" in The Biochemistry Global Summit, 9th-14th July, 2022. In: FEBS Open Bio, 12, no. Suppl. 1 (2022):302-303,
https://hdl.handle.net/21.15107/rcub_cer_6945 .

TY  - CONF
AU  - Nedić, Olgica
AU  - Gligorijević, Nikola
AU  - Penezić, Ana
AU  - Minić, Simeon
AU  - Radomirović, Mirjana
AU  - Nikolić, Milan
AU  - Ćirković Veličković, Tanja
PY  - 2022
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6728
AB  - Our research work was focused on interactions between resveratrol (R) and tibnnogen (I), (dihydro)alpha-lipoic acid (ALA) and fibrinogen or albumin, and phycocyanobilin (PCB) and catalase. Resveratrol is found in grapes and berries, leafy greens are a source of ALA and alga Spirulina is a source of PCB. L-P interactions were investigated by following-up structural changes of proteins and/or ligands using spectrometric methods (spectrofluorimetry, CD, FTIR) and by examining the primary role of individual proteins upon ligand binding.
PB  - Belgrade : University of Belgrade - Faculty of Agriculture
C3  - Book of abstracts - 1st European Symposium on Phytochemicals in Medicine and Food (1EuSPMF), 7-9 September 2022, Belgrade, Serbia
T1  - Food antioxidants and their interaction with human proteins
SP  - 13
EP  - 13
UR  - https://hdl.handle.net/21.15107/rcub_cer_6728
ER  - 

@conference{
author = "Nedić, Olgica and Gligorijević, Nikola and Penezić, Ana and Minić, Simeon and Radomirović, Mirjana and Nikolić, Milan and Ćirković Veličković, Tanja",
year = "2022",
abstract = "Our research work was focused on interactions between resveratrol (R) and tibnnogen (I), (dihydro)alpha-lipoic acid (ALA) and fibrinogen or albumin, and phycocyanobilin (PCB) and catalase. Resveratrol is found in grapes and berries, leafy greens are a source of ALA and alga Spirulina is a source of PCB. L-P interactions were investigated by following-up structural changes of proteins and/or ligands using spectrometric methods (spectrofluorimetry, CD, FTIR) and by examining the primary role of individual proteins upon ligand binding.",
publisher = "Belgrade : University of Belgrade - Faculty of Agriculture",
journal = "Book of abstracts - 1st European Symposium on Phytochemicals in Medicine and Food (1EuSPMF), 7-9 September 2022, Belgrade, Serbia",
title = "Food antioxidants and their interaction with human proteins",
pages = "13-13",
url = "https://hdl.handle.net/21.15107/rcub_cer_6728"
}

Nedić, O., Gligorijević, N., Penezić, A., Minić, S., Radomirović, M., Nikolić, M.,& Ćirković Veličković, T.. (2022). Food antioxidants and their interaction with human proteins. in Book of abstracts - 1st European Symposium on Phytochemicals in Medicine and Food (1EuSPMF), 7-9 September 2022, Belgrade, Serbia
Belgrade : University of Belgrade - Faculty of Agriculture., 13-13.
https://hdl.handle.net/21.15107/rcub_cer_6728

Nedić O, Gligorijević N, Penezić A, Minić S, Radomirović M, Nikolić M, Ćirković Veličković T. Food antioxidants and their interaction with human proteins. in Book of abstracts - 1st European Symposium on Phytochemicals in Medicine and Food (1EuSPMF), 7-9 September 2022, Belgrade, Serbia. 2022;:13-13.
https://hdl.handle.net/21.15107/rcub_cer_6728 .

Nedić, Olgica, Gligorijević, Nikola, Penezić, Ana, Minić, Simeon, Radomirović, Mirjana, Nikolić, Milan, Ćirković Veličković, Tanja, "Food antioxidants and their interaction with human proteins" in Book of abstracts - 1st European Symposium on Phytochemicals in Medicine and Food (1EuSPMF), 7-9 September 2022, Belgrade, Serbia (2022):13-13,
https://hdl.handle.net/21.15107/rcub_cer_6728 .

TY  - CONF
AU  - Radomirović, Mirjana
AU  - Gligorijević, Nikola
AU  - Rajković, Andreja
AU  - Ćirković Veličković, Tanja
PY  - 2022
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6799
AB  - Tropomyosin (TPM) is considered a major allergen among different shellfish species.
Developing sensitive, specific, and reliable methods for quantifying TPM in food products
is crucial for persons allergic to shellfish. We have previously developed a highly sensitive
sandwich ELISA method for quantifying shrimp TPM. Despite high amino acid sequence
homology between shrimp and mussels TPM, the method has not been reliable for
quantifying TPM from mussels, underestimating its concentration up to three orders of
magnitude. Therefore, this work aimed to develop alternative immunological methods for
mussel TPM quantification. Western blot, dot blot, and indirect ELISA using monoclonal
anti-TPM antibody and alkaline phosphatase-labeled secondary antibody were developed
and compared in terms of their sensitivity. Tropomyosin in mussels extracts was quantified
using highly purified natural shrimp tropomyosin as standard. The linear range for TPM
quantification using dot blot was between 5 and 50 μg/mL, while Western blot has slightly
increased sensitivity, with a linear range between 1.25 and 12.5 μg/mL. Indirect ELISA
has further improved the sensitivity of TPM quantification, with a 0.04-0.4 μg/mL linear
range. Additional work will be performed to enhance the sensitivity of the presented
methods, with the final aim of reducing risks of inadvertent food contamination.
PB  - Serbian Biochemical Society
PB  - University of Belgrade - Faculty of Chemistry
C3  - Proceedings - XI Conference of Serbian Biochemical Society "Amazing Biochemistry", 22.09.2022. Novi Sad, Serbia, 2022, 2022, 141- Publisher:
T1  - Development and comparison of Western blot, dot blot and ELISA for mussel tropomyosin quantification
SP  - 125
EP  - 125
UR  - https://hdl.handle.net/21.15107/rcub_cer_6799
ER  - 

@conference{
author = "Radomirović, Mirjana and Gligorijević, Nikola and Rajković, Andreja and Ćirković Veličković, Tanja",
year = "2022",
abstract = "Tropomyosin (TPM) is considered a major allergen among different shellfish species.
Developing sensitive, specific, and reliable methods for quantifying TPM in food products
is crucial for persons allergic to shellfish. We have previously developed a highly sensitive
sandwich ELISA method for quantifying shrimp TPM. Despite high amino acid sequence
homology between shrimp and mussels TPM, the method has not been reliable for
quantifying TPM from mussels, underestimating its concentration up to three orders of
magnitude. Therefore, this work aimed to develop alternative immunological methods for
mussel TPM quantification. Western blot, dot blot, and indirect ELISA using monoclonal
anti-TPM antibody and alkaline phosphatase-labeled secondary antibody were developed
and compared in terms of their sensitivity. Tropomyosin in mussels extracts was quantified
using highly purified natural shrimp tropomyosin as standard. The linear range for TPM
quantification using dot blot was between 5 and 50 μg/mL, while Western blot has slightly
increased sensitivity, with a linear range between 1.25 and 12.5 μg/mL. Indirect ELISA
has further improved the sensitivity of TPM quantification, with a 0.04-0.4 μg/mL linear
range. Additional work will be performed to enhance the sensitivity of the presented
methods, with the final aim of reducing risks of inadvertent food contamination.",
publisher = "Serbian Biochemical Society, University of Belgrade - Faculty of Chemistry",
journal = "Proceedings - XI Conference of Serbian Biochemical Society "Amazing Biochemistry", 22.09.2022. Novi Sad, Serbia, 2022, 2022, 141- Publisher:",
title = "Development and comparison of Western blot, dot blot and ELISA for mussel tropomyosin quantification",
pages = "125-125",
url = "https://hdl.handle.net/21.15107/rcub_cer_6799"
}

Radomirović, M., Gligorijević, N., Rajković, A.,& Ćirković Veličković, T.. (2022). Development and comparison of Western blot, dot blot and ELISA for mussel tropomyosin quantification. in Proceedings - XI Conference of Serbian Biochemical Society "Amazing Biochemistry", 22.09.2022. Novi Sad, Serbia, 2022, 2022, 141- Publisher:
Serbian Biochemical Society., 125-125.
https://hdl.handle.net/21.15107/rcub_cer_6799

Radomirović M, Gligorijević N, Rajković A, Ćirković Veličković T. Development and comparison of Western blot, dot blot and ELISA for mussel tropomyosin quantification. in Proceedings - XI Conference of Serbian Biochemical Society "Amazing Biochemistry", 22.09.2022. Novi Sad, Serbia, 2022, 2022, 141- Publisher:. 2022;:125-125.
https://hdl.handle.net/21.15107/rcub_cer_6799 .

Radomirović, Mirjana, Gligorijević, Nikola, Rajković, Andreja, Ćirković Veličković, Tanja, "Development and comparison of Western blot, dot blot and ELISA for mussel tropomyosin quantification" in Proceedings - XI Conference of Serbian Biochemical Society "Amazing Biochemistry", 22.09.2022. Novi Sad, Serbia, 2022, 2022, 141- Publisher: (2022):125-125,
https://hdl.handle.net/21.15107/rcub_cer_6799 .

TY  - CONF
AU  - Radomirović, Mirjana
AU  - Čolaković, Maša
AU  - Pismestrović, Marina
AU  - Gligorijević, Nikola
AU  - Stanić-Vučinić, Dragana
AU  - Rajković, Andreja
AU  - Ćirković Veličković, Tanja
PY  - 2022
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6823
AB  - Tropomyosin (TPM) is considered as a major allergen among different shellfish species.
Therefore, the development of methods for quantifying TPM in food products iscrucial for
allergic persons. Several extraction buffers were tested for their efficiency in recovering
proteins from fresh frozen and cooked razor mud shrimp during 2 and 24 hours of
extraction. The protein content was quantified using the Bradford protein assay. SDSPAGE
was used for protein profiling of soluble extracts. Sandwich ELISA was developed
and used to quantify TPM content. None of the extraction buffers showed a significant
difference in total protein content between 2 and 24 hours of extraction. Significantly
fewer proteins were extracted from cooked shrimp compared to the raw shrimp. ELISA
quantification showed that phosphate-buffered saline (PBS) containing 1 M NaCl (PBSN)
and carbonate buffer, pH 10, extracted approximately 6 times higher amount of
tropomyosin in comparison to PBS, highlighting the importance of choosing the
appropriate extraction buffer for the precise quantification of TPM. Traditionally used
extraction buffer PBS could significantly underestimate shrimp TPM content.
AB  - Tropomiozin (TPM) se smatra glavnim alergenom morskih plodova. Razvoj metoda za kvantifikaciju TPM-a u prehrambenim proizvodima je ključan za alergične osobe. Nekoliko pufera za ekstrakciju je upoređeno u pogledu efikasnosti ekstrakcije protein iz sveže zamrznutih i kuvanih gambora, tokom 2 i 24 časa. Sadržaj proteina u solubilnim ekstraktima je određen Bredfordovom metodom. Proteinski profil ekstrakata je analiziran SDS-PAGE metodom. Za kvantifikaciju TPM-a je razvijena sendvič ELISA metoda. Nijedan puffer nije pokazao značajnu razliku u količini ekstrahovanih protein nakon 2 i 24 časa ekstrakcije. U poređenju sa sirovim, značajno manje proteina je ekstrahovano iz kuvanih gambora. Kvantifikacija TPM-a ELISA metodom je pokazala da se fosfatom puferisanim fiziološkim rastvorom (PBS) koji sadrži 1 M NaCl (PBSN) i karbonatnim 
puferom, pH 10, ekstrahuje oko 6 puta više TPM-a u poređenju sa PBS-om. Ovo ukazuje 
na značaj odabira adekvatnog pufera za ekstrakciju TPM-a, jer se upotrebom tradicionalno
korišćenih pufera može potceniti koncentracija TPM-a u gamborima.
PB  - Belgrade : Serbian Chemical Society
C3  - 58th Meeting of  the Serbian Chemical Society - Book of abstracts, Proceedings,  June 9-10, 2022 Belgrade / 58. Savetovanje Srpskog hemijskog društva, Kratki izvodi radova, Knjiga radova, 9. i 10. jun 2022. godine, Beograd
T1  - Optimization of extraction conditions of tropomyosin from razor mud  shrimp and its quantification by developed ELISA
SP  - 64
EP  - 64
UR  - https://hdl.handle.net/21.15107/rcub_cer_6823
ER  - 

@conference{
author = "Radomirović, Mirjana and Čolaković, Maša and Pismestrović, Marina and Gligorijević, Nikola and Stanić-Vučinić, Dragana and Rajković, Andreja and Ćirković Veličković, Tanja",
year = "2022",
abstract = "Tropomyosin (TPM) is considered as a major allergen among different shellfish species.
Therefore, the development of methods for quantifying TPM in food products iscrucial for
allergic persons. Several extraction buffers were tested for their efficiency in recovering
proteins from fresh frozen and cooked razor mud shrimp during 2 and 24 hours of
extraction. The protein content was quantified using the Bradford protein assay. SDSPAGE
was used for protein profiling of soluble extracts. Sandwich ELISA was developed
and used to quantify TPM content. None of the extraction buffers showed a significant
difference in total protein content between 2 and 24 hours of extraction. Significantly
fewer proteins were extracted from cooked shrimp compared to the raw shrimp. ELISA
quantification showed that phosphate-buffered saline (PBS) containing 1 M NaCl (PBSN)
and carbonate buffer, pH 10, extracted approximately 6 times higher amount of
tropomyosin in comparison to PBS, highlighting the importance of choosing the
appropriate extraction buffer for the precise quantification of TPM. Traditionally used
extraction buffer PBS could significantly underestimate shrimp TPM content., Tropomiozin (TPM) se smatra glavnim alergenom morskih plodova. Razvoj metoda za kvantifikaciju TPM-a u prehrambenim proizvodima je ključan za alergične osobe. Nekoliko pufera za ekstrakciju je upoređeno u pogledu efikasnosti ekstrakcije protein iz sveže zamrznutih i kuvanih gambora, tokom 2 i 24 časa. Sadržaj proteina u solubilnim ekstraktima je određen Bredfordovom metodom. Proteinski profil ekstrakata je analiziran SDS-PAGE metodom. Za kvantifikaciju TPM-a je razvijena sendvič ELISA metoda. Nijedan puffer nije pokazao značajnu razliku u količini ekstrahovanih protein nakon 2 i 24 časa ekstrakcije. U poređenju sa sirovim, značajno manje proteina je ekstrahovano iz kuvanih gambora. Kvantifikacija TPM-a ELISA metodom je pokazala da se fosfatom puferisanim fiziološkim rastvorom (PBS) koji sadrži 1 M NaCl (PBSN) i karbonatnim 
puferom, pH 10, ekstrahuje oko 6 puta više TPM-a u poređenju sa PBS-om. Ovo ukazuje 
na značaj odabira adekvatnog pufera za ekstrakciju TPM-a, jer se upotrebom tradicionalno
korišćenih pufera može potceniti koncentracija TPM-a u gamborima.",
publisher = "Belgrade : Serbian Chemical Society",
journal = "58th Meeting of  the Serbian Chemical Society - Book of abstracts, Proceedings,  June 9-10, 2022 Belgrade / 58. Savetovanje Srpskog hemijskog društva, Kratki izvodi radova, Knjiga radova, 9. i 10. jun 2022. godine, Beograd",
title = "Optimization of extraction conditions of tropomyosin from razor mud  shrimp and its quantification by developed ELISA",
pages = "64-64",
url = "https://hdl.handle.net/21.15107/rcub_cer_6823"
}

Radomirović, M., Čolaković, M., Pismestrović, M., Gligorijević, N., Stanić-Vučinić, D., Rajković, A.,& Ćirković Veličković, T.. (2022). Optimization of extraction conditions of tropomyosin from razor mud  shrimp and its quantification by developed ELISA. in 58th Meeting of  the Serbian Chemical Society - Book of abstracts, Proceedings,  June 9-10, 2022 Belgrade / 58. Savetovanje Srpskog hemijskog društva, Kratki izvodi radova, Knjiga radova, 9. i 10. jun 2022. godine, Beograd
Belgrade : Serbian Chemical Society., 64-64.
https://hdl.handle.net/21.15107/rcub_cer_6823

Radomirović M, Čolaković M, Pismestrović M, Gligorijević N, Stanić-Vučinić D, Rajković A, Ćirković Veličković T. Optimization of extraction conditions of tropomyosin from razor mud  shrimp and its quantification by developed ELISA. in 58th Meeting of  the Serbian Chemical Society - Book of abstracts, Proceedings,  June 9-10, 2022 Belgrade / 58. Savetovanje Srpskog hemijskog društva, Kratki izvodi radova, Knjiga radova, 9. i 10. jun 2022. godine, Beograd. 2022;:64-64.
https://hdl.handle.net/21.15107/rcub_cer_6823 .

Radomirović, Mirjana, Čolaković, Maša, Pismestrović, Marina, Gligorijević, Nikola, Stanić-Vučinić, Dragana, Rajković, Andreja, Ćirković Veličković, Tanja, "Optimization of extraction conditions of tropomyosin from razor mud  shrimp and its quantification by developed ELISA" in 58th Meeting of  the Serbian Chemical Society - Book of abstracts, Proceedings,  June 9-10, 2022 Belgrade / 58. Savetovanje Srpskog hemijskog društva, Kratki izvodi radova, Knjiga radova, 9. i 10. jun 2022. godine, Beograd (2022):64-64,
https://hdl.handle.net/21.15107/rcub_cer_6823 .

TY  - JOUR
AU  - Radosavljević, Jelena
AU  - Stanić-Vučinić, Dragana
AU  - Stojadinović, Marija M.
AU  - Radomirović, Mirjana Ž.
AU  - Simović, Ana
AU  - Radibratović, Milica
AU  - Ćirković-Veličković, Tanja
PY  - 2022
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/5555
AB  - Background: The world’s production of whey is estimated to be more than 200 milliontons per year. Although whey is an important source of proteins with high nutritional value andbiotechnological importance, it is still considered as a by-product of the dairy industry with loweconomic value due to low industrial exploitation. There are several challenges in the separation ofwhey proteins: low concentration, the complexity of the material and similar properties (pI, molecularmass) of some proteins.Methods: A narrative review of all the relevant papers on the present methodologies based on ionexchange and adsorption principles for isolation of whey proteins, known to the authors, was conductedResults: Traditional ion exchange techniques are widely used for the separation and purification ofthe bovine whey proteins. These methodologies, based on the anion or cation chromatographicprocedures, as well as the combination of aforementioned techniques are still preferential methodsfor the isolation of the whey proteins on the laboratory scale. However, more recent research on ionexchange membranes for this purpose has been introduced, with promising potential to be appliedon the pilot industrial scale. Newly developed methodologies based either on the ion exchangeseparation (for example: simulated moving bed chromatography, expanded bed adsorption, magneticion exchangers, etc.) or adsorption (for example: adsorption on hydroxyapatite or activatedcarbon, or molecular imprinting) are promising approaches for scaling up of the whey proteins’purification processes.Conclusion: Many procedures based on ion exchange are successfully implemented for the separationand purification of whey proteins, providing protein preparations of moderate-to-high yieldand satisfactory purity. However, the authors anticipate further development of adsorption-basedmethodologies for the separation of whey proteins by targeting the differences in proteins’ structuresrather than targeting the differences in molecular masses and pI. The complex composite multilayeredmatrices, including also inorganic components, are promising materials for simultaneousexploiting of the differences in the masses, pI and structures of whey proteins for the separation.
PB  - Bentham Science
T2  - Current Analytical Chemistry
T1  - Application of Ion Exchange and Adsorption Techniques for Separation of Whey Proteins from Bovine Milk
VL  - 18
IS  - 3
SP  - 341
EP  - 359
DO  - 10.2174/1573411017666210108092338
ER  - 

@article{
author = "Radosavljević, Jelena and Stanić-Vučinić, Dragana and Stojadinović, Marija M. and Radomirović, Mirjana Ž. and Simović, Ana and Radibratović, Milica and Ćirković-Veličković, Tanja",
year = "2022",
abstract = "Background: The world’s production of whey is estimated to be more than 200 milliontons per year. Although whey is an important source of proteins with high nutritional value andbiotechnological importance, it is still considered as a by-product of the dairy industry with loweconomic value due to low industrial exploitation. There are several challenges in the separation ofwhey proteins: low concentration, the complexity of the material and similar properties (pI, molecularmass) of some proteins.Methods: A narrative review of all the relevant papers on the present methodologies based on ionexchange and adsorption principles for isolation of whey proteins, known to the authors, was conductedResults: Traditional ion exchange techniques are widely used for the separation and purification ofthe bovine whey proteins. These methodologies, based on the anion or cation chromatographicprocedures, as well as the combination of aforementioned techniques are still preferential methodsfor the isolation of the whey proteins on the laboratory scale. However, more recent research on ionexchange membranes for this purpose has been introduced, with promising potential to be appliedon the pilot industrial scale. Newly developed methodologies based either on the ion exchangeseparation (for example: simulated moving bed chromatography, expanded bed adsorption, magneticion exchangers, etc.) or adsorption (for example: adsorption on hydroxyapatite or activatedcarbon, or molecular imprinting) are promising approaches for scaling up of the whey proteins’purification processes.Conclusion: Many procedures based on ion exchange are successfully implemented for the separationand purification of whey proteins, providing protein preparations of moderate-to-high yieldand satisfactory purity. However, the authors anticipate further development of adsorption-basedmethodologies for the separation of whey proteins by targeting the differences in proteins’ structuresrather than targeting the differences in molecular masses and pI. The complex composite multilayeredmatrices, including also inorganic components, are promising materials for simultaneousexploiting of the differences in the masses, pI and structures of whey proteins for the separation.",
publisher = "Bentham Science",
journal = "Current Analytical Chemistry",
title = "Application of Ion Exchange and Adsorption Techniques for Separation of Whey Proteins from Bovine Milk",
volume = "18",
number = "3",
pages = "341-359",
doi = "10.2174/1573411017666210108092338"
}

Radosavljević, J., Stanić-Vučinić, D., Stojadinović, M. M., Radomirović, M. Ž., Simović, A., Radibratović, M.,& Ćirković-Veličković, T.. (2022). Application of Ion Exchange and Adsorption Techniques for Separation of Whey Proteins from Bovine Milk. in Current Analytical Chemistry
Bentham Science., 18(3), 341-359.
https://doi.org/10.2174/1573411017666210108092338

Radosavljević J, Stanić-Vučinić D, Stojadinović MM, Radomirović MŽ, Simović A, Radibratović M, Ćirković-Veličković T. Application of Ion Exchange and Adsorption Techniques for Separation of Whey Proteins from Bovine Milk. in Current Analytical Chemistry. 2022;18(3):341-359.
doi:10.2174/1573411017666210108092338 .

Radosavljević, Jelena, Stanić-Vučinić, Dragana, Stojadinović, Marija M., Radomirović, Mirjana Ž., Simović, Ana, Radibratović, Milica, Ćirković-Veličković, Tanja, "Application of Ion Exchange and Adsorption Techniques for Separation of Whey Proteins from Bovine Milk" in Current Analytical Chemistry, 18, no. 3 (2022):341-359,
https://doi.org/10.2174/1573411017666210108092338 . .

TY  - JOUR
AU  - Radosavljević, Jelena
AU  - Stanić-Vučinić, Dragana
AU  - Stojadinović, Marija M.
AU  - Radomirović, Mirjana Ž.
AU  - Simović, Ana
AU  - Radibratović, Milica
AU  - Ćirković-Veličković, Tanja
PY  - 2022
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/5556
AB  - Background: The world’s production of whey is estimated to be more than 200 milliontons per year. Although whey is an important source of proteins with high nutritional value andbiotechnological importance, it is still considered as a by-product of the dairy industry with loweconomic value due to low industrial exploitation. There are several challenges in the separation ofwhey proteins: low concentration, the complexity of the material and similar properties (pI, molecularmass) of some proteins.Methods: A narrative review of all the relevant papers on the present methodologies based on ionexchange and adsorption principles for isolation of whey proteins, known to the authors, was conductedResults: Traditional ion exchange techniques are widely used for the separation and purification ofthe bovine whey proteins. These methodologies, based on the anion or cation chromatographicprocedures, as well as the combination of aforementioned techniques are still preferential methodsfor the isolation of the whey proteins on the laboratory scale. However, more recent research on ionexchange membranes for this purpose has been introduced, with promising potential to be appliedon the pilot industrial scale. Newly developed methodologies based either on the ion exchangeseparation (for example: simulated moving bed chromatography, expanded bed adsorption, magneticion exchangers, etc.) or adsorption (for example: adsorption on hydroxyapatite or activatedcarbon, or molecular imprinting) are promising approaches for scaling up of the whey proteins’purification processes.Conclusion: Many procedures based on ion exchange are successfully implemented for the separationand purification of whey proteins, providing protein preparations of moderate-to-high yieldand satisfactory purity. However, the authors anticipate further development of adsorption-basedmethodologies for the separation of whey proteins by targeting the differences in proteins’ structuresrather than targeting the differences in molecular masses and pI. The complex composite multilayeredmatrices, including also inorganic components, are promising materials for simultaneousexploiting of the differences in the masses, pI and structures of whey proteins for the separation.
PB  - Bentham Science
T2  - Current Analytical Chemistry
T1  - Application of Ion Exchange and Adsorption Techniques for Separation of Whey Proteins from Bovine Milk
VL  - 18
IS  - 3
SP  - 341
EP  - 359
DO  - 10.2174/1573411017666210108092338
ER  - 

@article{
author = "Radosavljević, Jelena and Stanić-Vučinić, Dragana and Stojadinović, Marija M. and Radomirović, Mirjana Ž. and Simović, Ana and Radibratović, Milica and Ćirković-Veličković, Tanja",
year = "2022",
abstract = "Background: The world’s production of whey is estimated to be more than 200 milliontons per year. Although whey is an important source of proteins with high nutritional value andbiotechnological importance, it is still considered as a by-product of the dairy industry with loweconomic value due to low industrial exploitation. There are several challenges in the separation ofwhey proteins: low concentration, the complexity of the material and similar properties (pI, molecularmass) of some proteins.Methods: A narrative review of all the relevant papers on the present methodologies based on ionexchange and adsorption principles for isolation of whey proteins, known to the authors, was conductedResults: Traditional ion exchange techniques are widely used for the separation and purification ofthe bovine whey proteins. These methodologies, based on the anion or cation chromatographicprocedures, as well as the combination of aforementioned techniques are still preferential methodsfor the isolation of the whey proteins on the laboratory scale. However, more recent research on ionexchange membranes for this purpose has been introduced, with promising potential to be appliedon the pilot industrial scale. Newly developed methodologies based either on the ion exchangeseparation (for example: simulated moving bed chromatography, expanded bed adsorption, magneticion exchangers, etc.) or adsorption (for example: adsorption on hydroxyapatite or activatedcarbon, or molecular imprinting) are promising approaches for scaling up of the whey proteins’purification processes.Conclusion: Many procedures based on ion exchange are successfully implemented for the separationand purification of whey proteins, providing protein preparations of moderate-to-high yieldand satisfactory purity. However, the authors anticipate further development of adsorption-basedmethodologies for the separation of whey proteins by targeting the differences in proteins’ structuresrather than targeting the differences in molecular masses and pI. The complex composite multilayeredmatrices, including also inorganic components, are promising materials for simultaneousexploiting of the differences in the masses, pI and structures of whey proteins for the separation.",
publisher = "Bentham Science",
journal = "Current Analytical Chemistry",
title = "Application of Ion Exchange and Adsorption Techniques for Separation of Whey Proteins from Bovine Milk",
volume = "18",
number = "3",
pages = "341-359",
doi = "10.2174/1573411017666210108092338"
}

Radosavljević, J., Stanić-Vučinić, D., Stojadinović, M. M., Radomirović, M. Ž., Simović, A., Radibratović, M.,& Ćirković-Veličković, T.. (2022). Application of Ion Exchange and Adsorption Techniques for Separation of Whey Proteins from Bovine Milk. in Current Analytical Chemistry
Bentham Science., 18(3), 341-359.
https://doi.org/10.2174/1573411017666210108092338

Radosavljević J, Stanić-Vučinić D, Stojadinović MM, Radomirović MŽ, Simović A, Radibratović M, Ćirković-Veličković T. Application of Ion Exchange and Adsorption Techniques for Separation of Whey Proteins from Bovine Milk. in Current Analytical Chemistry. 2022;18(3):341-359.
doi:10.2174/1573411017666210108092338 .

Radosavljević, Jelena, Stanić-Vučinić, Dragana, Stojadinović, Marija M., Radomirović, Mirjana Ž., Simović, Ana, Radibratović, Milica, Ćirković-Veličković, Tanja, "Application of Ion Exchange and Adsorption Techniques for Separation of Whey Proteins from Bovine Milk" in Current Analytical Chemistry, 18, no. 3 (2022):341-359,
https://doi.org/10.2174/1573411017666210108092338 . .

TY  - CONF
AU  - Radomirović, Mirjana
AU  - Gligorijević, Nikola
AU  - Minić, Simeon
AU  - Nikolić, Milan
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković-Veličković, Tanja
PY  - 2021
UR  - https://ec.europa.eu/research/participants/documents/downloadPublic?documentIds=080166e5deeb546e&appId=PPGMS
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7282
AB  - Phycobiliproteins (PBP) have been employed in numerous fluorescence-based techniques owing
to highly fluorescent, covalently bound tetrapyrrole chromophores. So far, only entire PBPs have
been utilized as fluorescent probes. A new method for covalent attachment of phycocyanin’s
chromophore, phycocyanobilin (PCB), to potentially any protein, is proposed, relying on the
ability of PCB to be attached to sulfhydryl groups of proteins. Traut’s reagent (TR, 2-
iminothiolane) was exploited for introduction of sulfhydryl groups in the model protein, bovine
serum albumin (BSA), by modifying its primary amines. Introduced sulfhydryl groups were then
targeted for modification by PCB. All tested molar ratios of TR per mole of protein were
successful in modification of BSA. Near-UV and far-UV circular dichroism spectroscopy
revealed that a higher degree of modification by TR induces more profound alterations of BSA
structure, leading at the same time to minor changes in BSA oligomerization and aggregation
profile. PCB was covalently attached to introduced sulfhydryl groups at pH 9 at 20–fold ratio of
TR. An increase in the molar ratio of TR per mole of BSA leads to amplification of fluorescent
signal of PCB-modified BSA, most significantly observed starting from 50-fold and higher TR
ratios. Using BSA as a model protein, a 50-fold molar excess of TR seems to be the optimal ratio
for balancing between the effect on protein structure and the degree of labeling and thus
fluorescent signal obtained. The proposed method could be used for labeling of virtually any
protein, as means of either obtaining fluorescent probes for application in fluorescent techniques
or functionalization of, for example, food proteins through covalent attachment of bioactive
PCB.
PB  - University of Belgrade - Faculty of Chemistry
C3  - FoodEnTwin Symposium “Novel analytical approaches in food and environmental sciences”, Book of Abstracts, 16th-18th June, 2021, Belgrade, Serbia
T1  - Traut’s reagent application in fluorescent labeling of bovine serum albumin with phycocyanobilin
SP  - 37
EP  - 37
UR  - https://hdl.handle.net/21.15107/rcub_cer_7282
ER  - 

@conference{
author = "Radomirović, Mirjana and Gligorijević, Nikola and Minić, Simeon and Nikolić, Milan and Stanić-Vučinić, Dragana and Ćirković-Veličković, Tanja",
year = "2021",
abstract = "Phycobiliproteins (PBP) have been employed in numerous fluorescence-based techniques owing
to highly fluorescent, covalently bound tetrapyrrole chromophores. So far, only entire PBPs have
been utilized as fluorescent probes. A new method for covalent attachment of phycocyanin’s
chromophore, phycocyanobilin (PCB), to potentially any protein, is proposed, relying on the
ability of PCB to be attached to sulfhydryl groups of proteins. Traut’s reagent (TR, 2-
iminothiolane) was exploited for introduction of sulfhydryl groups in the model protein, bovine
serum albumin (BSA), by modifying its primary amines. Introduced sulfhydryl groups were then
targeted for modification by PCB. All tested molar ratios of TR per mole of protein were
successful in modification of BSA. Near-UV and far-UV circular dichroism spectroscopy
revealed that a higher degree of modification by TR induces more profound alterations of BSA
structure, leading at the same time to minor changes in BSA oligomerization and aggregation
profile. PCB was covalently attached to introduced sulfhydryl groups at pH 9 at 20–fold ratio of
TR. An increase in the molar ratio of TR per mole of BSA leads to amplification of fluorescent
signal of PCB-modified BSA, most significantly observed starting from 50-fold and higher TR
ratios. Using BSA as a model protein, a 50-fold molar excess of TR seems to be the optimal ratio
for balancing between the effect on protein structure and the degree of labeling and thus
fluorescent signal obtained. The proposed method could be used for labeling of virtually any
protein, as means of either obtaining fluorescent probes for application in fluorescent techniques
or functionalization of, for example, food proteins through covalent attachment of bioactive
PCB.",
publisher = "University of Belgrade - Faculty of Chemistry",
journal = "FoodEnTwin Symposium “Novel analytical approaches in food and environmental sciences”, Book of Abstracts, 16th-18th June, 2021, Belgrade, Serbia",
title = "Traut’s reagent application in fluorescent labeling of bovine serum albumin with phycocyanobilin",
pages = "37-37",
url = "https://hdl.handle.net/21.15107/rcub_cer_7282"
}

Radomirović, M., Gligorijević, N., Minić, S., Nikolić, M., Stanić-Vučinić, D.,& Ćirković-Veličković, T.. (2021). Traut’s reagent application in fluorescent labeling of bovine serum albumin with phycocyanobilin. in FoodEnTwin Symposium “Novel analytical approaches in food and environmental sciences”, Book of Abstracts, 16th-18th June, 2021, Belgrade, Serbia
University of Belgrade - Faculty of Chemistry., 37-37.
https://hdl.handle.net/21.15107/rcub_cer_7282

Radomirović M, Gligorijević N, Minić S, Nikolić M, Stanić-Vučinić D, Ćirković-Veličković T. Traut’s reagent application in fluorescent labeling of bovine serum albumin with phycocyanobilin. in FoodEnTwin Symposium “Novel analytical approaches in food and environmental sciences”, Book of Abstracts, 16th-18th June, 2021, Belgrade, Serbia. 2021;:37-37.
https://hdl.handle.net/21.15107/rcub_cer_7282 .

Radomirović, Mirjana, Gligorijević, Nikola, Minić, Simeon, Nikolić, Milan, Stanić-Vučinić, Dragana, Ćirković-Veličković, Tanja, "Traut’s reagent application in fluorescent labeling of bovine serum albumin with phycocyanobilin" in FoodEnTwin Symposium “Novel analytical approaches in food and environmental sciences”, Book of Abstracts, 16th-18th June, 2021, Belgrade, Serbia (2021):37-37,
https://hdl.handle.net/21.15107/rcub_cer_7282 .

TY  - CONF
AU  - Gligorijević, Nikola
AU  - Minić, Simeon
AU  - Radomirović, Mirjana
AU  - Lević, Steva
AU  - Ćirković Veličković, Tanja
AU  - Nikolić, Milan
AU  - Nedić, Olgica
PY  - 2021
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7287
AB  - Fibrinogen is a plasma protein most susceptible to oxidation. Through this chemical
modification, fibrinogen acquires thrombogenic characteristics in different
pathophysiological conditions. Increased carbonyl content and reduced porosity impair the
degradation of formed fibrin mediated by plasmin. Fibrinogen is capable of interacting
with many proteins, ions, and small molecules. These interactions can modify the
functions of this protein. The discovery of new binding partners that may protect
fibrinogen from harmful oxidation and, thus, preserve its normal function is essential.
Some of the newly detected interactions between fibrinogen and small, natural bioactive
molecules, together with the influence of these interactions on the structure and function of
fibrinogen, will be presented in this text.
PB  - Serbian Biochemical Society
C3  - Serbian Biochemical Society Tenth Conference with international participation, “Biochemical Insights into Molecular Mechanisms”, 24.09.2021. Kragujevac, Serbia
T1  - Ligand binding to fibrinogen influences its structure and function
SP  - 31
EP  - 31
UR  - https://hdl.handle.net/21.15107/rcub_cer_7287
ER  - 

@conference{
author = "Gligorijević, Nikola and Minić, Simeon and Radomirović, Mirjana and Lević, Steva and Ćirković Veličković, Tanja and Nikolić, Milan and Nedić, Olgica",
year = "2021",
abstract = "Fibrinogen is a plasma protein most susceptible to oxidation. Through this chemical
modification, fibrinogen acquires thrombogenic characteristics in different
pathophysiological conditions. Increased carbonyl content and reduced porosity impair the
degradation of formed fibrin mediated by plasmin. Fibrinogen is capable of interacting
with many proteins, ions, and small molecules. These interactions can modify the
functions of this protein. The discovery of new binding partners that may protect
fibrinogen from harmful oxidation and, thus, preserve its normal function is essential.
Some of the newly detected interactions between fibrinogen and small, natural bioactive
molecules, together with the influence of these interactions on the structure and function of
fibrinogen, will be presented in this text.",
publisher = "Serbian Biochemical Society",
journal = "Serbian Biochemical Society Tenth Conference with international participation, “Biochemical Insights into Molecular Mechanisms”, 24.09.2021. Kragujevac, Serbia",
title = "Ligand binding to fibrinogen influences its structure and function",
pages = "31-31",
url = "https://hdl.handle.net/21.15107/rcub_cer_7287"
}

Gligorijević, N., Minić, S., Radomirović, M., Lević, S., Ćirković Veličković, T., Nikolić, M.,& Nedić, O.. (2021). Ligand binding to fibrinogen influences its structure and function. in Serbian Biochemical Society Tenth Conference with international participation, “Biochemical Insights into Molecular Mechanisms”, 24.09.2021. Kragujevac, Serbia
Serbian Biochemical Society., 31-31.
https://hdl.handle.net/21.15107/rcub_cer_7287

Gligorijević N, Minić S, Radomirović M, Lević S, Ćirković Veličković T, Nikolić M, Nedić O. Ligand binding to fibrinogen influences its structure and function. in Serbian Biochemical Society Tenth Conference with international participation, “Biochemical Insights into Molecular Mechanisms”, 24.09.2021. Kragujevac, Serbia. 2021;:31-31.
https://hdl.handle.net/21.15107/rcub_cer_7287 .

Gligorijević, Nikola, Minić, Simeon, Radomirović, Mirjana, Lević, Steva, Ćirković Veličković, Tanja, Nikolić, Milan, Nedić, Olgica, "Ligand binding to fibrinogen influences its structure and function" in Serbian Biochemical Society Tenth Conference with international participation, “Biochemical Insights into Molecular Mechanisms”, 24.09.2021. Kragujevac, Serbia (2021):31-31,
https://hdl.handle.net/21.15107/rcub_cer_7287 .

TY  - CONF
AU  - Radomirović, Mirjana
AU  - Gligorijević, Nikola
AU  - Minić, Simeon
AU  - Nikolić, Milan
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković Veličković, Tanja
PY  - 2021
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7290
AB  - Phycobiliproteins (PBP) are extensively used as fluorescent probes due to highly fluorescent,
covalently bound, tetrapyrrole chromophores. A new method for covalent attachment of
phycocyanin’s chromophore, phycocyanobilin (PCB), is proposed. We exploited Traut’s reagent
(TR) to introduce sulfhydryl groups in the bovine serum albumin (BSA), by modifying its lysine
residues. TR successfully modified BSA under all tested molar ratios of reagent per mole of BSA.
The higher degree of modification by TR induces more profound alterations of BSA structure. PCB
was covalently attached to introduced sulfhydryl groups at pH 9 at 20–fold ratio. An increase in the
molar ratio of TR per mole of BSA leads to amplification of fluorescent signal of PCB-modified
BSA. Using BSA as a model protein, a 50-fold molar excess of TR seems to be the optimal choice
for balancing between a satisfactory level of signal amplification and the adverse effect on protein
structure. The method could be used for labeling virtually any protein.
PB  - Belgrade : Serbian Chemical Society
C3  - Kratki izvodi radova, Knjiga radova 57. Savetovanje Srpskog hemijskog društva, 18. i 19. juni 2021, Kragujevac / Book of abstracts, Proceedings - 57th Meeting of the Serbian Chemical Society, June 18-19, 2021, Kragujevac, Serbia
T1  - Fluorescent labeling of bovine serum albumin with phycocyanobilin using Traut’s reagent
SP  - 71
EP  - 71
UR  - https://hdl.handle.net/21.15107/rcub_cer_7290
ER  - 

@conference{
author = "Radomirović, Mirjana and Gligorijević, Nikola and Minić, Simeon and Nikolić, Milan and Stanić-Vučinić, Dragana and Ćirković Veličković, Tanja",
year = "2021",
abstract = "Phycobiliproteins (PBP) are extensively used as fluorescent probes due to highly fluorescent,
covalently bound, tetrapyrrole chromophores. A new method for covalent attachment of
phycocyanin’s chromophore, phycocyanobilin (PCB), is proposed. We exploited Traut’s reagent
(TR) to introduce sulfhydryl groups in the bovine serum albumin (BSA), by modifying its lysine
residues. TR successfully modified BSA under all tested molar ratios of reagent per mole of BSA.
The higher degree of modification by TR induces more profound alterations of BSA structure. PCB
was covalently attached to introduced sulfhydryl groups at pH 9 at 20–fold ratio. An increase in the
molar ratio of TR per mole of BSA leads to amplification of fluorescent signal of PCB-modified
BSA. Using BSA as a model protein, a 50-fold molar excess of TR seems to be the optimal choice
for balancing between a satisfactory level of signal amplification and the adverse effect on protein
structure. The method could be used for labeling virtually any protein.",
publisher = "Belgrade : Serbian Chemical Society",
journal = "Kratki izvodi radova, Knjiga radova 57. Savetovanje Srpskog hemijskog društva, 18. i 19. juni 2021, Kragujevac / Book of abstracts, Proceedings - 57th Meeting of the Serbian Chemical Society, June 18-19, 2021, Kragujevac, Serbia",
title = "Fluorescent labeling of bovine serum albumin with phycocyanobilin using Traut’s reagent",
pages = "71-71",
url = "https://hdl.handle.net/21.15107/rcub_cer_7290"
}

Radomirović, M., Gligorijević, N., Minić, S., Nikolić, M., Stanić-Vučinić, D.,& Ćirković Veličković, T.. (2021). Fluorescent labeling of bovine serum albumin with phycocyanobilin using Traut’s reagent. in Kratki izvodi radova, Knjiga radova 57. Savetovanje Srpskog hemijskog društva, 18. i 19. juni 2021, Kragujevac / Book of abstracts, Proceedings - 57th Meeting of the Serbian Chemical Society, June 18-19, 2021, Kragujevac, Serbia
Belgrade : Serbian Chemical Society., 71-71.
https://hdl.handle.net/21.15107/rcub_cer_7290

Radomirović M, Gligorijević N, Minić S, Nikolić M, Stanić-Vučinić D, Ćirković Veličković T. Fluorescent labeling of bovine serum albumin with phycocyanobilin using Traut’s reagent. in Kratki izvodi radova, Knjiga radova 57. Savetovanje Srpskog hemijskog društva, 18. i 19. juni 2021, Kragujevac / Book of abstracts, Proceedings - 57th Meeting of the Serbian Chemical Society, June 18-19, 2021, Kragujevac, Serbia. 2021;:71-71.
https://hdl.handle.net/21.15107/rcub_cer_7290 .

Radomirović, Mirjana, Gligorijević, Nikola, Minić, Simeon, Nikolić, Milan, Stanić-Vučinić, Dragana, Ćirković Veličković, Tanja, "Fluorescent labeling of bovine serum albumin with phycocyanobilin using Traut’s reagent" in Kratki izvodi radova, Knjiga radova 57. Savetovanje Srpskog hemijskog društva, 18. i 19. juni 2021, Kragujevac / Book of abstracts, Proceedings - 57th Meeting of the Serbian Chemical Society, June 18-19, 2021, Kragujevac, Serbia (2021):71-71,
https://hdl.handle.net/21.15107/rcub_cer_7290 .

TY  - JOUR
AU  - Gligorijević, Nikola
AU  - Minić, Simeon
AU  - Radomirović, Mirjana
AU  - Lević, Steva M.
AU  - Nikolić, Milan
AU  - Ćirković-Veličković, Tanja
AU  - Nedić, Olgica
PY  - 2021
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7288
AB  - Fibrinogen is a plasma protein that is highly susceptible to oxidation. Because of this chemical modification, fibrinogen acquires thrombogenic characteristics under different pathophysiological conditions. Increased carbonyl content and reduced porosity impair the degradation of formed fibrin mediated by plasmin. Fibrinogen is capable of interacting with many proteins, ions, and small molecules. These interactions can modify the functions of this protein. The discovery of new binding partners that may protect fibrinogen from harmful oxidation and, thus, preserve its normal function is essential. Some of the newly detected interactions between fibrinogen and small, natural bioactive molecules, together with the influence of these interactions on the structure and function of fibrinogen, will be presented in this text.
PB  - University of Novi Sad - Faculty of Sciences, Department of Biology
T2  - Biologia Serbica
T1  - Ligand binding to fibrinogen influences its structure and function
VL  - 43
IS  - 1
SP  - 24
EP  - 31
DO  - 10.5281/zenodo.5512285
ER  - 

@article{
author = "Gligorijević, Nikola and Minić, Simeon and Radomirović, Mirjana and Lević, Steva M. and Nikolić, Milan and Ćirković-Veličković, Tanja and Nedić, Olgica",
year = "2021",
abstract = "Fibrinogen is a plasma protein that is highly susceptible to oxidation. Because of this chemical modification, fibrinogen acquires thrombogenic characteristics under different pathophysiological conditions. Increased carbonyl content and reduced porosity impair the degradation of formed fibrin mediated by plasmin. Fibrinogen is capable of interacting with many proteins, ions, and small molecules. These interactions can modify the functions of this protein. The discovery of new binding partners that may protect fibrinogen from harmful oxidation and, thus, preserve its normal function is essential. Some of the newly detected interactions between fibrinogen and small, natural bioactive molecules, together with the influence of these interactions on the structure and function of fibrinogen, will be presented in this text.",
publisher = "University of Novi Sad - Faculty of Sciences, Department of Biology",
journal = "Biologia Serbica",
title = "Ligand binding to fibrinogen influences its structure and function",
volume = "43",
number = "1",
pages = "24-31",
doi = "10.5281/zenodo.5512285"
}

Gligorijević, N., Minić, S., Radomirović, M., Lević, S. M., Nikolić, M., Ćirković-Veličković, T.,& Nedić, O.. (2021). Ligand binding to fibrinogen influences its structure and function. in Biologia Serbica
University of Novi Sad - Faculty of Sciences, Department of Biology., 43(1), 24-31.
https://doi.org/10.5281/zenodo.5512285

Gligorijević N, Minić S, Radomirović M, Lević SM, Nikolić M, Ćirković-Veličković T, Nedić O. Ligand binding to fibrinogen influences its structure and function. in Biologia Serbica. 2021;43(1):24-31.
doi:10.5281/zenodo.5512285 .

Gligorijević, Nikola, Minić, Simeon, Radomirović, Mirjana, Lević, Steva M., Nikolić, Milan, Ćirković-Veličković, Tanja, Nedić, Olgica, "Ligand binding to fibrinogen influences its structure and function" in Biologia Serbica, 43, no. 1 (2021):24-31,
https://doi.org/10.5281/zenodo.5512285 . .

TY  - CONF
AU  - Radomirović, Mirjana
AU  - Gligorijević, Nikola
AU  - Stanić-Vučinić, Dragana
AU  - Rajković, Andreja
AU  - Ćirković Veličković, Tanja
PY  - 2021
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7280
AB  - Food allergies affect up to 10% of the general population and represent an important health problem in the field of
food safety in industrialized countries. Hence, developing reliable, specific, and sensitive methods for detecting and
quantifying allergens in food products is of high importance. Shellfish have been recognized as one of the eight
most common sources of allergens, with tropomyosin (TPM) being considered a major heat-stable allergen, having
a highly conserved amino acid sequence among different shellfish species. Allergenicity of TPM may change during
food processing, such as cooking. The objective of this study was to develop an enzyme-linked immunosorbent
assay (ELISA) for the detection and quantification of shellfish tropomyosin in food samples.
Two different extraction buffers - phosphate-buffered saline (PBS) and PBS containing 1 M sodium-chloride
(PBSN), were compared for their ability to recover proteins from pre-cooked frozen Mediterranean mussel (Mytilus
galloprovincialis) and fresh frozen razor mud shrimp (Solenocera melantho). The samples were additionally cooked
according to the manufacturer's instruction and analyzed as such. The protein content was quantified using Bradford
protein assay, and the protein components of soluble extracts were profiled using SDS-PAGE. TPM presence was
confirmed using Western blot. Sandwich ELISA was developed using a monoclonal anti-TPM antibody as a capture
antibody, while polyclonal anti-TPM antibody served as a detection antibody and was coupled to the biotinylated
secondary antibody and streptavidin-alkaline phosphatase conjugate. Tropomyosin was quantified using highly
purified natural shrimp tropomyosin as standard.
The profile of extracted proteins was changed when using PBSN instead of PBS. A higher concentration of proteins
was recovered from raw shrimp using PBSN instead of PBS. At the same time, the type of extraction buffer did not
affect protein recovery either from heated shrimp or pre-cooked/heated mussels. Significantly fewer proteins were
extracted from cooked shrimp sample compared to the raw shrimp, while cooking showed no effect on the extraction
of proteins from mussels. Cooking did not affect TPM recognition in Western blot. TPM was quantified in shrimp
samples in sandwich ELISA. However, developed ELISA could not quantify mussel's TPM, indicating that this
approach may distinguish mussels and shrimp TPM.
PB  - Sociedade Portuguesa de Química
C3  - Book of Abstracts - XXI EuroFoodChem conference, 22nd-24th November, 2021, Virtual Congress
T1  - Extraction and quantification of tropomyosin in selected samples of shellfish
SP  - 118
EP  - 118
UR  - https://hdl.handle.net/21.15107/rcub_cer_7280
ER  - 

@conference{
author = "Radomirović, Mirjana and Gligorijević, Nikola and Stanić-Vučinić, Dragana and Rajković, Andreja and Ćirković Veličković, Tanja",
year = "2021",
abstract = "Food allergies affect up to 10% of the general population and represent an important health problem in the field of
food safety in industrialized countries. Hence, developing reliable, specific, and sensitive methods for detecting and
quantifying allergens in food products is of high importance. Shellfish have been recognized as one of the eight
most common sources of allergens, with tropomyosin (TPM) being considered a major heat-stable allergen, having
a highly conserved amino acid sequence among different shellfish species. Allergenicity of TPM may change during
food processing, such as cooking. The objective of this study was to develop an enzyme-linked immunosorbent
assay (ELISA) for the detection and quantification of shellfish tropomyosin in food samples.
Two different extraction buffers - phosphate-buffered saline (PBS) and PBS containing 1 M sodium-chloride
(PBSN), were compared for their ability to recover proteins from pre-cooked frozen Mediterranean mussel (Mytilus
galloprovincialis) and fresh frozen razor mud shrimp (Solenocera melantho). The samples were additionally cooked
according to the manufacturer's instruction and analyzed as such. The protein content was quantified using Bradford
protein assay, and the protein components of soluble extracts were profiled using SDS-PAGE. TPM presence was
confirmed using Western blot. Sandwich ELISA was developed using a monoclonal anti-TPM antibody as a capture
antibody, while polyclonal anti-TPM antibody served as a detection antibody and was coupled to the biotinylated
secondary antibody and streptavidin-alkaline phosphatase conjugate. Tropomyosin was quantified using highly
purified natural shrimp tropomyosin as standard.
The profile of extracted proteins was changed when using PBSN instead of PBS. A higher concentration of proteins
was recovered from raw shrimp using PBSN instead of PBS. At the same time, the type of extraction buffer did not
affect protein recovery either from heated shrimp or pre-cooked/heated mussels. Significantly fewer proteins were
extracted from cooked shrimp sample compared to the raw shrimp, while cooking showed no effect on the extraction
of proteins from mussels. Cooking did not affect TPM recognition in Western blot. TPM was quantified in shrimp
samples in sandwich ELISA. However, developed ELISA could not quantify mussel's TPM, indicating that this
approach may distinguish mussels and shrimp TPM.",
publisher = "Sociedade Portuguesa de Química",
journal = "Book of Abstracts - XXI EuroFoodChem conference, 22nd-24th November, 2021, Virtual Congress",
title = "Extraction and quantification of tropomyosin in selected samples of shellfish",
pages = "118-118",
url = "https://hdl.handle.net/21.15107/rcub_cer_7280"
}

Radomirović, M., Gligorijević, N., Stanić-Vučinić, D., Rajković, A.,& Ćirković Veličković, T.. (2021). Extraction and quantification of tropomyosin in selected samples of shellfish. in Book of Abstracts - XXI EuroFoodChem conference, 22nd-24th November, 2021, Virtual Congress
Sociedade Portuguesa de Química., 118-118.
https://hdl.handle.net/21.15107/rcub_cer_7280

Radomirović M, Gligorijević N, Stanić-Vučinić D, Rajković A, Ćirković Veličković T. Extraction and quantification of tropomyosin in selected samples of shellfish. in Book of Abstracts - XXI EuroFoodChem conference, 22nd-24th November, 2021, Virtual Congress. 2021;:118-118.
https://hdl.handle.net/21.15107/rcub_cer_7280 .

Radomirović, Mirjana, Gligorijević, Nikola, Stanić-Vučinić, Dragana, Rajković, Andreja, Ćirković Veličković, Tanja, "Extraction and quantification of tropomyosin in selected samples of shellfish" in Book of Abstracts - XXI EuroFoodChem conference, 22nd-24th November, 2021, Virtual Congress (2021):118-118,
https://hdl.handle.net/21.15107/rcub_cer_7280 .

TY  - CONF
AU  - Gligorijević, Nikola
AU  - Radomirović, Mirjana
AU  - Rajković, Andreja
AU  - Nedić, Olgica
AU  - Ćirković Veličković, Tanja
PY  - 2021
UR  - https://ec.europa.eu/research/participants/documents/downloadPublic?documentIds=080166e5deeb546e&appId=PPGMS
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7284
AB  - The French paradox describes a lower incidence of cardiovascular problems despite a high intake of
saturated fats. This phenomenon was associated with higher consumption of red wine, only to be later discovered that the presence of several antioxidants, including resveratrol, are responsible for it. We investigated if resveratrol has a more direct role in protection from harmful oxidation and development of thrombosis, presumably through binding to important proteins of the blood coagulation process. Spectrofluorimetric analysis demonstrated binding of resveratrol to fibrinogen, the main protein in the coagulation process, which also has an important application as a food additive in making of fibrin gels. Various spectroscopic methods have demonstrated that binding of
resveratrol does not unfold or destabilize fibrinogen since both near and far-UV CD spectra as well as its melting temperature remained unchanged. A mutuallyprotective effect against the free radical-
induced oxidation of resveratrol and fibrinogen was found. The presence of fibrinogen caused a very
small masking effect of the antioxidative potential of resveratrol, measured by a reduction of
hexacyanoferrate (III), while greatly increasing its solubility in an aqueous environment, thus increasing potential bioavailability and activity of resveratrol in circulation. By direct interaction and protection of fibrinogen, resveratrol may serve as an important antioxidant for prevention of
thrombosis. The antioxidative effect of resveratrol may also protect and thus keep the desired characteristics of fibrinogen during the application of this protein as a food additive.
PB  - University of Belgrade - Faculty of Chemistry
C3  - FoodEnTwin Symposium “Novel analytical approaches in food and environmental sciences”, Book of Abstracts, 16th-18th June, 2021, Belgrade, Serbia
T1  - Resveratrol and fibrinogen interactions
SP  - 15
EP  - 15
UR  - https://hdl.handle.net/21.15107/rcub_cer_7284
ER  - 

@conference{
author = "Gligorijević, Nikola and Radomirović, Mirjana and Rajković, Andreja and Nedić, Olgica and Ćirković Veličković, Tanja",
year = "2021",
abstract = "The French paradox describes a lower incidence of cardiovascular problems despite a high intake of
saturated fats. This phenomenon was associated with higher consumption of red wine, only to be later discovered that the presence of several antioxidants, including resveratrol, are responsible for it. We investigated if resveratrol has a more direct role in protection from harmful oxidation and development of thrombosis, presumably through binding to important proteins of the blood coagulation process. Spectrofluorimetric analysis demonstrated binding of resveratrol to fibrinogen, the main protein in the coagulation process, which also has an important application as a food additive in making of fibrin gels. Various spectroscopic methods have demonstrated that binding of
resveratrol does not unfold or destabilize fibrinogen since both near and far-UV CD spectra as well as its melting temperature remained unchanged. A mutuallyprotective effect against the free radical-
induced oxidation of resveratrol and fibrinogen was found. The presence of fibrinogen caused a very
small masking effect of the antioxidative potential of resveratrol, measured by a reduction of
hexacyanoferrate (III), while greatly increasing its solubility in an aqueous environment, thus increasing potential bioavailability and activity of resveratrol in circulation. By direct interaction and protection of fibrinogen, resveratrol may serve as an important antioxidant for prevention of
thrombosis. The antioxidative effect of resveratrol may also protect and thus keep the desired characteristics of fibrinogen during the application of this protein as a food additive.",
publisher = "University of Belgrade - Faculty of Chemistry",
journal = "FoodEnTwin Symposium “Novel analytical approaches in food and environmental sciences”, Book of Abstracts, 16th-18th June, 2021, Belgrade, Serbia",
title = "Resveratrol and fibrinogen interactions",
pages = "15-15",
url = "https://hdl.handle.net/21.15107/rcub_cer_7284"
}

Gligorijević, N., Radomirović, M., Rajković, A., Nedić, O.,& Ćirković Veličković, T.. (2021). Resveratrol and fibrinogen interactions. in FoodEnTwin Symposium “Novel analytical approaches in food and environmental sciences”, Book of Abstracts, 16th-18th June, 2021, Belgrade, Serbia
University of Belgrade - Faculty of Chemistry., 15-15.
https://hdl.handle.net/21.15107/rcub_cer_7284

Gligorijević N, Radomirović M, Rajković A, Nedić O, Ćirković Veličković T. Resveratrol and fibrinogen interactions. in FoodEnTwin Symposium “Novel analytical approaches in food and environmental sciences”, Book of Abstracts, 16th-18th June, 2021, Belgrade, Serbia. 2021;:15-15.
https://hdl.handle.net/21.15107/rcub_cer_7284 .

Gligorijević, Nikola, Radomirović, Mirjana, Rajković, Andreja, Nedić, Olgica, Ćirković Veličković, Tanja, "Resveratrol and fibrinogen interactions" in FoodEnTwin Symposium “Novel analytical approaches in food and environmental sciences”, Book of Abstracts, 16th-18th June, 2021, Belgrade, Serbia (2021):15-15,
https://hdl.handle.net/21.15107/rcub_cer_7284 .

TY  - JOUR
AU  - Radibratović, Milica
AU  - Al-Hanish, Ayah
AU  - Minić, Simeon
AU  - Radomirović, Mirjana
AU  - Milčić, Miloš
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković Veličković, Tanja
PY  - 2019
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/3212
AB  - α-Lactalbumin (ALA) is a Ca2+-binding protein which constitutes up to 20% of whey protein. At acidic pH, and in the apo-state at elevated temperatures, ALA is the classic 'molten globule' (MG). This study examined epigallocatechin-3-gallate (EGCG) binding to ALA in its apo form (apoALA) and stabilizing effect on protein structure thereof.

EGCG binds to apoALA in both native and MG state. The complex of EGCG and ALA is more stable to thermal denaturation. The docking analysis and molecular dynamic simulation (MDS) showed that Ca2+ removal decreased conformational stability of ALA, because of the local destabilization of Ca2+-binding region. EGCG binding to apoALA increases its stability by reverting of conformation and stability of Ca2+-binding region. Therefore, EGCG-induced thermal stability of apoALA is based on increased apoALA conformational rigidity. This study implies that during gastric digestion of tea with milk EGCG would remain bound to ALA, albeit in the Ca2+-free form
PB  - Elsevier
T2  - Food Chemistry
T1  - Stabilization of apo α-lactalbumin by binding of epigallocatechin-3-gallate: Experimental and molecular dynamics study
VL  - 278
SP  - 388
EP  - 395
DO  - 10.1016/j.foodchem.2018.11.038
ER  - 

@article{
author = "Radibratović, Milica and Al-Hanish, Ayah and Minić, Simeon and Radomirović, Mirjana and Milčić, Miloš and Stanić-Vučinić, Dragana and Ćirković Veličković, Tanja",
year = "2019",
abstract = "α-Lactalbumin (ALA) is a Ca2+-binding protein which constitutes up to 20% of whey protein. At acidic pH, and in the apo-state at elevated temperatures, ALA is the classic 'molten globule' (MG). This study examined epigallocatechin-3-gallate (EGCG) binding to ALA in its apo form (apoALA) and stabilizing effect on protein structure thereof.

EGCG binds to apoALA in both native and MG state. The complex of EGCG and ALA is more stable to thermal denaturation. The docking analysis and molecular dynamic simulation (MDS) showed that Ca2+ removal decreased conformational stability of ALA, because of the local destabilization of Ca2+-binding region. EGCG binding to apoALA increases its stability by reverting of conformation and stability of Ca2+-binding region. Therefore, EGCG-induced thermal stability of apoALA is based on increased apoALA conformational rigidity. This study implies that during gastric digestion of tea with milk EGCG would remain bound to ALA, albeit in the Ca2+-free form",
publisher = "Elsevier",
journal = "Food Chemistry",
title = "Stabilization of apo α-lactalbumin by binding of epigallocatechin-3-gallate: Experimental and molecular dynamics study",
volume = "278",
pages = "388-395",
doi = "10.1016/j.foodchem.2018.11.038"
}

Radibratović, M., Al-Hanish, A., Minić, S., Radomirović, M., Milčić, M., Stanić-Vučinić, D.,& Ćirković Veličković, T.. (2019). Stabilization of apo α-lactalbumin by binding of epigallocatechin-3-gallate: Experimental and molecular dynamics study. in Food Chemistry
Elsevier., 278, 388-395.
https://doi.org/10.1016/j.foodchem.2018.11.038

Radibratović M, Al-Hanish A, Minić S, Radomirović M, Milčić M, Stanić-Vučinić D, Ćirković Veličković T. Stabilization of apo α-lactalbumin by binding of epigallocatechin-3-gallate: Experimental and molecular dynamics study. in Food Chemistry. 2019;278:388-395.
doi:10.1016/j.foodchem.2018.11.038 .

Radibratović, Milica, Al-Hanish, Ayah, Minić, Simeon, Radomirović, Mirjana, Milčić, Miloš, Stanić-Vučinić, Dragana, Ćirković Veličković, Tanja, "Stabilization of apo α-lactalbumin by binding of epigallocatechin-3-gallate: Experimental and molecular dynamics study" in Food Chemistry, 278 (2019):388-395,
https://doi.org/10.1016/j.foodchem.2018.11.038 . .

TY  - JOUR
AU  - Minić, Simeon
AU  - Radomirović, Mirjana
AU  - Savkovic, Nina
AU  - Radibratović, Milica
AU  - Mihailović, Jelena
AU  - Vasović, Tamara
AU  - Nikolić, Milan
AU  - Milčić, Miloš
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković Veličković, Tanja
PY  - 2018
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/2420
AB  - In this study, we investigated structural aspects of covalent binding of food derived blue pigment phycocyanobilin (PCB) to bovine beta-lactoglobulin (BLG), major whey protein, by spectroscopic, electrophoretic, mass spectrometry and computational methods. At physiological pH (7.2), we found that covalent pigment binding via free cysteine residue is slow (k(a)=0.065 min(-1)), of moderate affinity (K-a=4x10(4) M-1), and stereo-selective. Binding also occurs at a broad pH range and under simulated gastrointestinal conditions. Adduct formation rises with pH, and in concentrated urea (k(a)=0.101 min(-1)). The BLG-PCB adduct has slightly altered secondary and tertiary protein structure, and bound PCB has higher fluorescence and more stretched conformation than free chromophore. Combination of steered molecular dynamic for disulfide exchange, non-covalent and covalent docking, favours Cys119 residue in protein calyx as target for covalent BLG-PCB adduct formation. Our results suggest that this adduct can serve as delivery system of bioactive PCB.
PB  - Elsevier Sci Ltd, Oxford
T2  - Food Chemistry
T1  - Covalent binding of food-derived blue pigment phycocyanobilin to bovine beta-lactoglobulin under physiological conditions
VL  - 269
SP  - 43
EP  - 52
DO  - 10.1016/j.foodchem.2018.06.138
ER  - 

@article{
author = "Minić, Simeon and Radomirović, Mirjana and Savkovic, Nina and Radibratović, Milica and Mihailović, Jelena and Vasović, Tamara and Nikolić, Milan and Milčić, Miloš and Stanić-Vučinić, Dragana and Ćirković Veličković, Tanja",
year = "2018",
abstract = "In this study, we investigated structural aspects of covalent binding of food derived blue pigment phycocyanobilin (PCB) to bovine beta-lactoglobulin (BLG), major whey protein, by spectroscopic, electrophoretic, mass spectrometry and computational methods. At physiological pH (7.2), we found that covalent pigment binding via free cysteine residue is slow (k(a)=0.065 min(-1)), of moderate affinity (K-a=4x10(4) M-1), and stereo-selective. Binding also occurs at a broad pH range and under simulated gastrointestinal conditions. Adduct formation rises with pH, and in concentrated urea (k(a)=0.101 min(-1)). The BLG-PCB adduct has slightly altered secondary and tertiary protein structure, and bound PCB has higher fluorescence and more stretched conformation than free chromophore. Combination of steered molecular dynamic for disulfide exchange, non-covalent and covalent docking, favours Cys119 residue in protein calyx as target for covalent BLG-PCB adduct formation. Our results suggest that this adduct can serve as delivery system of bioactive PCB.",
publisher = "Elsevier Sci Ltd, Oxford",
journal = "Food Chemistry",
title = "Covalent binding of food-derived blue pigment phycocyanobilin to bovine beta-lactoglobulin under physiological conditions",
volume = "269",
pages = "43-52",
doi = "10.1016/j.foodchem.2018.06.138"
}

Minić, S., Radomirović, M., Savkovic, N., Radibratović, M., Mihailović, J., Vasović, T., Nikolić, M., Milčić, M., Stanić-Vučinić, D.,& Ćirković Veličković, T.. (2018). Covalent binding of food-derived blue pigment phycocyanobilin to bovine beta-lactoglobulin under physiological conditions. in Food Chemistry
Elsevier Sci Ltd, Oxford., 269, 43-52.
https://doi.org/10.1016/j.foodchem.2018.06.138

Minić S, Radomirović M, Savkovic N, Radibratović M, Mihailović J, Vasović T, Nikolić M, Milčić M, Stanić-Vučinić D, Ćirković Veličković T. Covalent binding of food-derived blue pigment phycocyanobilin to bovine beta-lactoglobulin under physiological conditions. in Food Chemistry. 2018;269:43-52.
doi:10.1016/j.foodchem.2018.06.138 .

Minić, Simeon, Radomirović, Mirjana, Savkovic, Nina, Radibratović, Milica, Mihailović, Jelena, Vasović, Tamara, Nikolić, Milan, Milčić, Miloš, Stanić-Vučinić, Dragana, Ćirković Veličković, Tanja, "Covalent binding of food-derived blue pigment phycocyanobilin to bovine beta-lactoglobulin under physiological conditions" in Food Chemistry, 269 (2018):43-52,
https://doi.org/10.1016/j.foodchem.2018.06.138 . .

TY  - JOUR
AU  - Minić, Simeon
AU  - Stanić-Vučinić, Dragana
AU  - Radomirović, Mirjana
AU  - Radibratović, Milica
AU  - Milčić, Miloš
AU  - Nikolić, Milan
AU  - Ćirković Veličković, Tanja
PY  - 2018
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/2478
AB  - Phycocyanobilin (PCB) is a blue tetrapyrrole chromophore of C-phycocyanin, the main protein of the microalga Spirulina, with numerous proven health-related benefits. We examined binding of PCB to bovine serum albumin (BSA) and how it affects protein and ligand stability. Protein fluorescence quenching and microscale thermophoresis demonstrated high-affinity binding (K-a = 2 x 10(6) M-1). Spectroscopic titration with molecular docking analysis revealed two binding sites on BSA, at the inter-domain cleft and at subdomain IB, while CD spectroscopy indicated stereo-selective binding of the P conformer of the pigment to the protein. The PCB protein complex showed increased thermal stability. Although complex formation partly masked the antioxidant properties of PCB and BSA, a mutually protective effect against free radical-induced oxidation was found. BSA could be suitable for delivery of PCB as a food colorant or bioactive component. Our results also highlight subtle differences between PCB binding to bovine vs. human serum albumin.
PB  - Elsevier Sci Ltd, Oxford
T2  - Food Chemistry
T1  - Characterization and effects of binding of food-derived bioactive phycocyanobilin to bovine serum albumin
VL  - 239
SP  - 1090
EP  - 1099
DO  - 10.1016/j.foodchem.2017.07.066
ER  - 

@article{
author = "Minić, Simeon and Stanić-Vučinić, Dragana and Radomirović, Mirjana and Radibratović, Milica and Milčić, Miloš and Nikolić, Milan and Ćirković Veličković, Tanja",
year = "2018",
abstract = "Phycocyanobilin (PCB) is a blue tetrapyrrole chromophore of C-phycocyanin, the main protein of the microalga Spirulina, with numerous proven health-related benefits. We examined binding of PCB to bovine serum albumin (BSA) and how it affects protein and ligand stability. Protein fluorescence quenching and microscale thermophoresis demonstrated high-affinity binding (K-a = 2 x 10(6) M-1). Spectroscopic titration with molecular docking analysis revealed two binding sites on BSA, at the inter-domain cleft and at subdomain IB, while CD spectroscopy indicated stereo-selective binding of the P conformer of the pigment to the protein. The PCB protein complex showed increased thermal stability. Although complex formation partly masked the antioxidant properties of PCB and BSA, a mutually protective effect against free radical-induced oxidation was found. BSA could be suitable for delivery of PCB as a food colorant or bioactive component. Our results also highlight subtle differences between PCB binding to bovine vs. human serum albumin.",
publisher = "Elsevier Sci Ltd, Oxford",
journal = "Food Chemistry",
title = "Characterization and effects of binding of food-derived bioactive phycocyanobilin to bovine serum albumin",
volume = "239",
pages = "1090-1099",
doi = "10.1016/j.foodchem.2017.07.066"
}

Minić, S., Stanić-Vučinić, D., Radomirović, M., Radibratović, M., Milčić, M., Nikolić, M.,& Ćirković Veličković, T.. (2018). Characterization and effects of binding of food-derived bioactive phycocyanobilin to bovine serum albumin. in Food Chemistry
Elsevier Sci Ltd, Oxford., 239, 1090-1099.
https://doi.org/10.1016/j.foodchem.2017.07.066

Minić S, Stanić-Vučinić D, Radomirović M, Radibratović M, Milčić M, Nikolić M, Ćirković Veličković T. Characterization and effects of binding of food-derived bioactive phycocyanobilin to bovine serum albumin. in Food Chemistry. 2018;239:1090-1099.
doi:10.1016/j.foodchem.2017.07.066 .

Minić, Simeon, Stanić-Vučinić, Dragana, Radomirović, Mirjana, Radibratović, Milica, Milčić, Miloš, Nikolić, Milan, Ćirković Veličković, Tanja, "Characterization and effects of binding of food-derived bioactive phycocyanobilin to bovine serum albumin" in Food Chemistry, 239 (2018):1090-1099,
https://doi.org/10.1016/j.foodchem.2017.07.066 . .

TY  - JOUR
AU  - Minić, Simeon
AU  - Stanić-Vučinić, Dragana
AU  - Radomirović, Mirjana
AU  - Radibratović, Milica
AU  - Milčić, Miloš
AU  - Nikolić, Milan
AU  - Ćirković Veličković, Tanja
PY  - 2018
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/2984
AB  - Phycocyanobilin (PCB) is a blue tetrapyrrole chromophore of C-phycocyanin, the main protein of the microalga Spirulina, with numerous proven health-related benefits. We examined binding of PCB to bovine serum albumin (BSA) and how it affects protein and ligand stability. Protein fluorescence quenching and microscale thermophoresis demonstrated high-affinity binding (K-a = 2 x 10(6) M-1). Spectroscopic titration with molecular docking analysis revealed two binding sites on BSA, at the inter-domain cleft and at subdomain IB, while CD spectroscopy indicated stereo-selective binding of the P conformer of the pigment to the protein. The PCB protein complex showed increased thermal stability. Although complex formation partly masked the antioxidant properties of PCB and BSA, a mutually protective effect against free radical-induced oxidation was found. BSA could be suitable for delivery of PCB as a food colorant or bioactive component. Our results also highlight subtle differences between PCB binding to bovine vs. human serum albumin.
PB  - Elsevier Sci Ltd, Oxford
T2  - Food Chemistry
T1  - Characterization and effects of binding of food-derived bioactive phycocyanobilin to bovine serum albumin
VL  - 239
SP  - 1090
EP  - 1099
DO  - 10.1016/j.foodchem.2017.07.066
ER  - 

@article{
author = "Minić, Simeon and Stanić-Vučinić, Dragana and Radomirović, Mirjana and Radibratović, Milica and Milčić, Miloš and Nikolić, Milan and Ćirković Veličković, Tanja",
year = "2018",
abstract = "Phycocyanobilin (PCB) is a blue tetrapyrrole chromophore of C-phycocyanin, the main protein of the microalga Spirulina, with numerous proven health-related benefits. We examined binding of PCB to bovine serum albumin (BSA) and how it affects protein and ligand stability. Protein fluorescence quenching and microscale thermophoresis demonstrated high-affinity binding (K-a = 2 x 10(6) M-1). Spectroscopic titration with molecular docking analysis revealed two binding sites on BSA, at the inter-domain cleft and at subdomain IB, while CD spectroscopy indicated stereo-selective binding of the P conformer of the pigment to the protein. The PCB protein complex showed increased thermal stability. Although complex formation partly masked the antioxidant properties of PCB and BSA, a mutually protective effect against free radical-induced oxidation was found. BSA could be suitable for delivery of PCB as a food colorant or bioactive component. Our results also highlight subtle differences between PCB binding to bovine vs. human serum albumin.",
publisher = "Elsevier Sci Ltd, Oxford",
journal = "Food Chemistry",
title = "Characterization and effects of binding of food-derived bioactive phycocyanobilin to bovine serum albumin",
volume = "239",
pages = "1090-1099",
doi = "10.1016/j.foodchem.2017.07.066"
}

Minić, S., Stanić-Vučinić, D., Radomirović, M., Radibratović, M., Milčić, M., Nikolić, M.,& Ćirković Veličković, T.. (2018). Characterization and effects of binding of food-derived bioactive phycocyanobilin to bovine serum albumin. in Food Chemistry
Elsevier Sci Ltd, Oxford., 239, 1090-1099.
https://doi.org/10.1016/j.foodchem.2017.07.066

Minić S, Stanić-Vučinić D, Radomirović M, Radibratović M, Milčić M, Nikolić M, Ćirković Veličković T. Characterization and effects of binding of food-derived bioactive phycocyanobilin to bovine serum albumin. in Food Chemistry. 2018;239:1090-1099.
doi:10.1016/j.foodchem.2017.07.066 .

Minić, Simeon, Stanić-Vučinić, Dragana, Radomirović, Mirjana, Radibratović, Milica, Milčić, Miloš, Nikolić, Milan, Ćirković Veličković, Tanja, "Characterization and effects of binding of food-derived bioactive phycocyanobilin to bovine serum albumin" in Food Chemistry, 239 (2018):1090-1099,
https://doi.org/10.1016/j.foodchem.2017.07.066 . .

TY  - JOUR
AU  - Minić, Simeon
AU  - Radomirović, Mirjana
AU  - Savkovic, Nina
AU  - Radibratović, Milica
AU  - Mihailović, Jelena
AU  - Vasović, Tamara
AU  - Nikolić, Milan
AU  - Milčić, Miloš
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković Veličković, Tanja
PY  - 2018
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/3022
AB  - In this study, we investigated structural aspects of covalent binding of food derived blue pigment phycocyanobilin (PCB) to bovine beta-lactoglobulin (BLG), major whey protein, by spectroscopic, electrophoretic, mass spectrometry and computational methods. At physiological pH (7.2), we found that covalent pigment binding via free cysteine residue is slow (k(a)=0.065 min(-1)), of moderate affinity (K-a=4x10(4) M-1), and stereo-selective. Binding also occurs at a broad pH range and under simulated gastrointestinal conditions. Adduct formation rises with pH, and in concentrated urea (k(a)=0.101 min(-1)). The BLG-PCB adduct has slightly altered secondary and tertiary protein structure, and bound PCB has higher fluorescence and more stretched conformation than free chromophore. Combination of steered molecular dynamic for disulfide exchange, non-covalent and covalent docking, favours Cys119 residue in protein calyx as target for covalent BLG-PCB adduct formation. Our results suggest that this adduct can serve as delivery system of bioactive PCB.
PB  - Elsevier Sci Ltd, Oxford
T2  - Food Chemistry
T1  - Covalent binding of food-derived blue pigment phycocyanobilin to bovine beta-lactoglobulin under physiological conditions
VL  - 269
SP  - 43
EP  - 52
DO  - 10.1016/j.foodchem.2018.06.138
ER  - 

@article{
author = "Minić, Simeon and Radomirović, Mirjana and Savkovic, Nina and Radibratović, Milica and Mihailović, Jelena and Vasović, Tamara and Nikolić, Milan and Milčić, Miloš and Stanić-Vučinić, Dragana and Ćirković Veličković, Tanja",
year = "2018",
abstract = "In this study, we investigated structural aspects of covalent binding of food derived blue pigment phycocyanobilin (PCB) to bovine beta-lactoglobulin (BLG), major whey protein, by spectroscopic, electrophoretic, mass spectrometry and computational methods. At physiological pH (7.2), we found that covalent pigment binding via free cysteine residue is slow (k(a)=0.065 min(-1)), of moderate affinity (K-a=4x10(4) M-1), and stereo-selective. Binding also occurs at a broad pH range and under simulated gastrointestinal conditions. Adduct formation rises with pH, and in concentrated urea (k(a)=0.101 min(-1)). The BLG-PCB adduct has slightly altered secondary and tertiary protein structure, and bound PCB has higher fluorescence and more stretched conformation than free chromophore. Combination of steered molecular dynamic for disulfide exchange, non-covalent and covalent docking, favours Cys119 residue in protein calyx as target for covalent BLG-PCB adduct formation. Our results suggest that this adduct can serve as delivery system of bioactive PCB.",
publisher = "Elsevier Sci Ltd, Oxford",
journal = "Food Chemistry",
title = "Covalent binding of food-derived blue pigment phycocyanobilin to bovine beta-lactoglobulin under physiological conditions",
volume = "269",
pages = "43-52",
doi = "10.1016/j.foodchem.2018.06.138"
}

Minić, S., Radomirović, M., Savkovic, N., Radibratović, M., Mihailović, J., Vasović, T., Nikolić, M., Milčić, M., Stanić-Vučinić, D.,& Ćirković Veličković, T.. (2018). Covalent binding of food-derived blue pigment phycocyanobilin to bovine beta-lactoglobulin under physiological conditions. in Food Chemistry
Elsevier Sci Ltd, Oxford., 269, 43-52.
https://doi.org/10.1016/j.foodchem.2018.06.138

Minić S, Radomirović M, Savkovic N, Radibratović M, Mihailović J, Vasović T, Nikolić M, Milčić M, Stanić-Vučinić D, Ćirković Veličković T. Covalent binding of food-derived blue pigment phycocyanobilin to bovine beta-lactoglobulin under physiological conditions. in Food Chemistry. 2018;269:43-52.
doi:10.1016/j.foodchem.2018.06.138 .

Minić, Simeon, Radomirović, Mirjana, Savkovic, Nina, Radibratović, Milica, Mihailović, Jelena, Vasović, Tamara, Nikolić, Milan, Milčić, Miloš, Stanić-Vučinić, Dragana, Ćirković Veličković, Tanja, "Covalent binding of food-derived blue pigment phycocyanobilin to bovine beta-lactoglobulin under physiological conditions" in Food Chemistry, 269 (2018):43-52,
https://doi.org/10.1016/j.foodchem.2018.06.138 . .

TY  - CONF
AU  - Radibratović, Milica
AU  - Al-Hanish, Ayah
AU  - Minić, Simeon
AU  - Radomirović, Mirjana Ž.
AU  - Milčić, Miloš
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković-Veličković, Tanja
PY  - 2018
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7462
AB  - α-Lactalbumin (ALA) is a small, acidic, Ca2+-binding protein that might constitute up to 20% of whey protein. ALA has several partially folded intermediate states. It is very attractive for studies of the properties and structure of intermediate molten globule-like states since at acidic pH and in the apo-state at elevated temperatures ALA is the classic ‘molten globule’.The aim of our study was to examine if epi-gallo-catechin-3-gallate (EGCG) binds to ALA in its Ca-depleted (apo-ALA) and less ordered states. Moreover we examined if the polyphenol binding can stabilize protein structure. The experimental investigation of EGCG binding and protein’s stability were supported by molecular dynamics data.Green tea catechin binds to apo-ALA in neutral and acidic conditions. Binding affinity determined by intrinsic fluorescence quenching is of a similar magnitude as to the holo form of the protein. The complex of EGCG and ALA at neutral conditions is more stable to thermal denaturation then protein alone. The stabilizing effect is more pronounced for apo-ALA than for the holo-ALA.The docking analysis and molecular dynamic simulation (MDS) showed that EGCG binds to apoALA at the same site as to holoALA, in the hydrophobic pocket at the entrance of cleft between α and β subdomains, remaining bound during MDS. Ca2+ removal results in decreased conformational stability of ALA, where local destabilization of Ca2+-binding region propagates to cleft, with its slight opening. EGCG binding to apo-ALA increases stability of ALA by reverting of conformation and stability of disturbed Ca2+-binding region, which is transmitted to cleft, resulting in rejoining of α and β subdomains and slight cleft closing by the same mechanism. EGCG binding could retard transition of apo-ALA to less ordered states under conditions favoring its formation.Therefore, non-covalent binding of EGCG can stabilize structural fold of calcium-depleted ALA.
PB  - Univerzitet u Beogradu
C3  - UNIFOOD Conference, Belgrade, Serbia, 5th–6th October, 2018. In: UNIFOOD Conference - Programme and Book of Abstracts
T1  - Stabilization of apo-alpha-lactalbumin by binding of epigallocatechin-3-gallate: experimental and molecular dynamics study
SP  - 268
EP  - 268
UR  - https://hdl.handle.net/21.15107/rcub_cer_7462
ER  - 

@conference{
author = "Radibratović, Milica and Al-Hanish, Ayah and Minić, Simeon and Radomirović, Mirjana Ž. and Milčić, Miloš and Stanić-Vučinić, Dragana and Ćirković-Veličković, Tanja",
year = "2018",
abstract = "α-Lactalbumin (ALA) is a small, acidic, Ca2+-binding protein that might constitute up to 20% of whey protein. ALA has several partially folded intermediate states. It is very attractive for studies of the properties and structure of intermediate molten globule-like states since at acidic pH and in the apo-state at elevated temperatures ALA is the classic ‘molten globule’.The aim of our study was to examine if epi-gallo-catechin-3-gallate (EGCG) binds to ALA in its Ca-depleted (apo-ALA) and less ordered states. Moreover we examined if the polyphenol binding can stabilize protein structure. The experimental investigation of EGCG binding and protein’s stability were supported by molecular dynamics data.Green tea catechin binds to apo-ALA in neutral and acidic conditions. Binding affinity determined by intrinsic fluorescence quenching is of a similar magnitude as to the holo form of the protein. The complex of EGCG and ALA at neutral conditions is more stable to thermal denaturation then protein alone. The stabilizing effect is more pronounced for apo-ALA than for the holo-ALA.The docking analysis and molecular dynamic simulation (MDS) showed that EGCG binds to apoALA at the same site as to holoALA, in the hydrophobic pocket at the entrance of cleft between α and β subdomains, remaining bound during MDS. Ca2+ removal results in decreased conformational stability of ALA, where local destabilization of Ca2+-binding region propagates to cleft, with its slight opening. EGCG binding to apo-ALA increases stability of ALA by reverting of conformation and stability of disturbed Ca2+-binding region, which is transmitted to cleft, resulting in rejoining of α and β subdomains and slight cleft closing by the same mechanism. EGCG binding could retard transition of apo-ALA to less ordered states under conditions favoring its formation.Therefore, non-covalent binding of EGCG can stabilize structural fold of calcium-depleted ALA.",
publisher = "Univerzitet u Beogradu",
journal = "UNIFOOD Conference, Belgrade, Serbia, 5th–6th October, 2018. In: UNIFOOD Conference - Programme and Book of Abstracts",
title = "Stabilization of apo-alpha-lactalbumin by binding of epigallocatechin-3-gallate: experimental and molecular dynamics study",
pages = "268-268",
url = "https://hdl.handle.net/21.15107/rcub_cer_7462"
}

Radibratović, M., Al-Hanish, A., Minić, S., Radomirović, M. Ž., Milčić, M., Stanić-Vučinić, D.,& Ćirković-Veličković, T.. (2018). Stabilization of apo-alpha-lactalbumin by binding of epigallocatechin-3-gallate: experimental and molecular dynamics study. in UNIFOOD Conference, Belgrade, Serbia, 5th–6th October, 2018. In: UNIFOOD Conference - Programme and Book of Abstracts
Univerzitet u Beogradu., 268-268.
https://hdl.handle.net/21.15107/rcub_cer_7462

Radibratović M, Al-Hanish A, Minić S, Radomirović MŽ, Milčić M, Stanić-Vučinić D, Ćirković-Veličković T. Stabilization of apo-alpha-lactalbumin by binding of epigallocatechin-3-gallate: experimental and molecular dynamics study. in UNIFOOD Conference, Belgrade, Serbia, 5th–6th October, 2018. In: UNIFOOD Conference - Programme and Book of Abstracts. 2018;:268-268.
https://hdl.handle.net/21.15107/rcub_cer_7462 .

Radibratović, Milica, Al-Hanish, Ayah, Minić, Simeon, Radomirović, Mirjana Ž., Milčić, Miloš, Stanić-Vučinić, Dragana, Ćirković-Veličković, Tanja, "Stabilization of apo-alpha-lactalbumin by binding of epigallocatechin-3-gallate: experimental and molecular dynamics study" in UNIFOOD Conference, Belgrade, Serbia, 5th–6th October, 2018. In: UNIFOOD Conference - Programme and Book of Abstracts (2018):268-268,
https://hdl.handle.net/21.15107/rcub_cer_7462 .

TY  - CONF
AU  - Minic, S.
AU  - Radomirović, Mirjana
AU  - Radibratović, Milica
AU  - Milčić, Miloš
AU  - Stanić-Vučinić, Dragana
AU  - Nikolić, Milan
AU  - Ćirković Veličković, Tanja
PY  - 2017
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/2167
UR  - https://doi.org/10.1111/febs.14174
PB  - Wiley, Hoboken
C3  - FEBS Journal
T1  - Characterization and effects of binding of food-derived bioactive phycocyanobilin to bovine serum albumin
VL  - 284
IS  - S1
SP  - 189
EP  - 190
UR  - https://hdl.handle.net/21.15107/rcub_cer_2167
ER  - 

@conference{
author = "Minic, S. and Radomirović, Mirjana and Radibratović, Milica and Milčić, Miloš and Stanić-Vučinić, Dragana and Nikolić, Milan and Ćirković Veličković, Tanja",
year = "2017",
publisher = "Wiley, Hoboken",
journal = "FEBS Journal",
title = "Characterization and effects of binding of food-derived bioactive phycocyanobilin to bovine serum albumin",
volume = "284",
number = "S1",
pages = "189-190",
url = "https://hdl.handle.net/21.15107/rcub_cer_2167"
}

Minic, S., Radomirović, M., Radibratović, M., Milčić, M., Stanić-Vučinić, D., Nikolić, M.,& Ćirković Veličković, T.. (2017). Characterization and effects of binding of food-derived bioactive phycocyanobilin to bovine serum albumin. in FEBS Journal
Wiley, Hoboken., 284(S1), 189-190.
https://hdl.handle.net/21.15107/rcub_cer_2167

Minic S, Radomirović M, Radibratović M, Milčić M, Stanić-Vučinić D, Nikolić M, Ćirković Veličković T. Characterization and effects of binding of food-derived bioactive phycocyanobilin to bovine serum albumin. in FEBS Journal. 2017;284(S1):189-190.
https://hdl.handle.net/21.15107/rcub_cer_2167 .

Minic, S., Radomirović, Mirjana, Radibratović, Milica, Milčić, Miloš, Stanić-Vučinić, Dragana, Nikolić, Milan, Ćirković Veličković, Tanja, "Characterization and effects of binding of food-derived bioactive phycocyanobilin to bovine serum albumin" in FEBS Journal, 284, no. S1 (2017):189-190,
https://hdl.handle.net/21.15107/rcub_cer_2167 .