TY  - JOUR
AU  - Guo, Lili
AU  - Zhao, Jing
AU  - An, Zongfu
AU  - Kim, Sieun
AU  - Kim, Jaekwang
AU  - Yu, Yeseul
AU  - Middelberg, Anton
AU  - Bi, Jingxiu
AU  - Marković, Marijana
AU  - Kim, Jung Kyu
AU  - Yoo, Pil J.
AU  - Choe, Woo-Seok
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7228
AB  - Despite the rapid advances in process analytical technology, the assessment of protein refolding efficiency has largely relied on off-line protein-specific assays and/or chromatographic procedures such as reversed-phase high-performance liquid chromatography and size exclusion chromatography. Due to the inherent time gap pertaining to traditional methods, exploring optimum refolding conditions for many recombinant proteins, often expressed as insoluble inclusion bodies, has proven challenging. The present study describes a novel protein refolding sensor that utilizes liquid crystals (LCs) to discriminate varying protein structures during unfolding and refolding. An LC layer containing 4-cyano-4′-pentylbiphenyl (5CB) intercalated with 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) is used as a sensing platform, and its proof-of-concept performance is demonstrated using lysozyme as a model protein. As proteins unfold or refold, a local charge fluctuation at their surfaces modulates their interaction with zwitterionic phospholipid DOPE. This alters the alignment of DOPE molecules at the aqueous/LC interface, affecting the orientational ordering of bulk LC (i.e., homeotropic to planar for refolding and planar to homeotropic for unfolding). Differential polarized optical microscope images of the LC layer are subsequently generated, whose brightness directly linked to conformational changes of lysozyme molecules is quantified by gray scale analysis. Importantly, our LC-based refolding sensor is compatible with diverse refolding milieus for real-time analysis of lysozyme refolding and thus likely to facilitate the refolding studies of many proteins, especially those lacking a method to determine structure-dependent biological activity.
PB  - American Chemical Society (ACS)
T2  - Analytical Chemistry
T1  - Harnessing Liquid Crystal Sensors for High-Throughput Real-Time Detection of Structural Changes in Lysozyme during Refolding Processes
VL  - 95
IS  - 48
SP  - 17603
EP  - 17612
DO  - 10.1021/acs.analchem.3c03272
ER  - 

@article{
author = "Guo, Lili and Zhao, Jing and An, Zongfu and Kim, Sieun and Kim, Jaekwang and Yu, Yeseul and Middelberg, Anton and Bi, Jingxiu and Marković, Marijana and Kim, Jung Kyu and Yoo, Pil J. and Choe, Woo-Seok",
year = "2023",
abstract = "Despite the rapid advances in process analytical technology, the assessment of protein refolding efficiency has largely relied on off-line protein-specific assays and/or chromatographic procedures such as reversed-phase high-performance liquid chromatography and size exclusion chromatography. Due to the inherent time gap pertaining to traditional methods, exploring optimum refolding conditions for many recombinant proteins, often expressed as insoluble inclusion bodies, has proven challenging. The present study describes a novel protein refolding sensor that utilizes liquid crystals (LCs) to discriminate varying protein structures during unfolding and refolding. An LC layer containing 4-cyano-4′-pentylbiphenyl (5CB) intercalated with 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) is used as a sensing platform, and its proof-of-concept performance is demonstrated using lysozyme as a model protein. As proteins unfold or refold, a local charge fluctuation at their surfaces modulates their interaction with zwitterionic phospholipid DOPE. This alters the alignment of DOPE molecules at the aqueous/LC interface, affecting the orientational ordering of bulk LC (i.e., homeotropic to planar for refolding and planar to homeotropic for unfolding). Differential polarized optical microscope images of the LC layer are subsequently generated, whose brightness directly linked to conformational changes of lysozyme molecules is quantified by gray scale analysis. Importantly, our LC-based refolding sensor is compatible with diverse refolding milieus for real-time analysis of lysozyme refolding and thus likely to facilitate the refolding studies of many proteins, especially those lacking a method to determine structure-dependent biological activity.",
publisher = "American Chemical Society (ACS)",
journal = "Analytical Chemistry",
title = "Harnessing Liquid Crystal Sensors for High-Throughput Real-Time Detection of Structural Changes in Lysozyme during Refolding Processes",
volume = "95",
number = "48",
pages = "17603-17612",
doi = "10.1021/acs.analchem.3c03272"
}

Guo, L., Zhao, J., An, Z., Kim, S., Kim, J., Yu, Y., Middelberg, A., Bi, J., Marković, M., Kim, J. K., Yoo, P. J.,& Choe, W.. (2023). Harnessing Liquid Crystal Sensors for High-Throughput Real-Time Detection of Structural Changes in Lysozyme during Refolding Processes. in Analytical Chemistry
American Chemical Society (ACS)., 95(48), 17603-17612.
https://doi.org/10.1021/acs.analchem.3c03272

Guo L, Zhao J, An Z, Kim S, Kim J, Yu Y, Middelberg A, Bi J, Marković M, Kim JK, Yoo PJ, Choe W. Harnessing Liquid Crystal Sensors for High-Throughput Real-Time Detection of Structural Changes in Lysozyme during Refolding Processes. in Analytical Chemistry. 2023;95(48):17603-17612.
doi:10.1021/acs.analchem.3c03272 .

Guo, Lili, Zhao, Jing, An, Zongfu, Kim, Sieun, Kim, Jaekwang, Yu, Yeseul, Middelberg, Anton, Bi, Jingxiu, Marković, Marijana, Kim, Jung Kyu, Yoo, Pil J., Choe, Woo-Seok, "Harnessing Liquid Crystal Sensors for High-Throughput Real-Time Detection of Structural Changes in Lysozyme during Refolding Processes" in Analytical Chemistry, 95, no. 48 (2023):17603-17612,
https://doi.org/10.1021/acs.analchem.3c03272 . .