Mechanistic insights into inter-chain disulfide bond reduction of IgG1 and IgG4 antibodies
| العنوان: | Mechanistic insights into inter-chain disulfide bond reduction of IgG1 and IgG4 antibodies |
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| المؤلفون: | Yuanli Song, Hui Cai, Zhijun Tan, Nesredin Mussa, Zheng-Jian Li |
| المصدر: | Applied Microbiology and Biotechnology. 106:1057-1066 |
| بيانات النشر: | Springer Science and Business Media LLC, 2022. |
| سنة النشر: | 2022 |
| مصطلحات موضوعية: | Immunoglobulin G, Antibodies, Bispecific, Antibodies, Monoclonal, Humans, Cysteine, Disulfides, General Medicine, Applied Microbiology and Biotechnology, Biotechnology |
| الوصف: | Therapeutic monoclonal antibodies (mAbs), primarily immunoglobin G1 (IgG1) and IgG4 with an engineered CPPC motif in its hinge region, are predominant biologics. Inter-chain disulfide bonds of IgG mAbs are crucial to maintaining IgG integrity. Inter-chain disulfide bond-reduced low molecular weight (LMW) is considered as one of quality attributes of IgG drug substance and is observed in drug substance manufacturing. In this study, we demonstrate that IgG1 and IgG4 are susceptible to the reducing agent TCEP differently and they follow different pathways to form LMWs. Our study shows that IgG1 is more sensitive to TCEP than IgG4. Both therapeutic IgG1 and human blood plasma IgG1 follow a heavy-heavy-light chain (HHL) pathway, featured with HHL and HH as intermediate species. Human blood plasma IgG4 with a CPSC motif in its hinge region follows heavy-light chain (HL) pathway, featured with HL as the intermediate species. However, therapeutic IgG4 follows a hybrid pathway with the HL pathway as the primary and the HHL pathway as the secondary. These experimental observations are further explained using solvent accessibility of inter-chain disulfide bonds obtained from computational modeling and molecular dynamics simulations. Findings from this study provide mechanistic insights of LMW formation of IgG1 and IgG4, which suggest selection of IgG1 or IgG4 for bispecific antibodies and cysteine-based antibody-drug conjugates. KEY POINTS: • Experimentally discovered preferable disulfide bond reduction pathways between IgG1 and IgG4 antibodies, driven by the different solvent accessibilities of these disulfide bonds. • Computationally explained the solvent accessibility aided by molecular dynamics simulations. • Provided insights in developing robust biologics process and designing bispecific antibodies and cysteine-based antibody-drug conjugates. |
| تدمد: | 1432-0614 0175-7598 |
| DOI: | 10.1007/s00253-022-11778-5 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::19950ff3820d2fa4d591a727c7e355e1 https://doi.org/10.1007/s00253-022-11778-5 |
| Rights: | CLOSED |
| رقم الانضمام: | edsair.doi.dedup.....19950ff3820d2fa4d591a727c7e355e1 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 14320614 01757598 |
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| DOI: | 10.1007/s00253-022-11778-5 |