Cargando…
Improving antibody affinity through in vitro mutagenesis in complementarity determining regions
High-affinity antibodies are widely used in diagnostics and for the treatment of human diseases. However, most antibodies are isolated from semi-synthetic libraries by phage display and do not possess in vivo affinity maturation, which is triggered by antigen immunization. It is therefore necessary...
Autores principales: | , , , , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Editorial Department of Journal of Biomedical Research
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9179109/ https://www.ncbi.nlm.nih.gov/pubmed/35545451 http://dx.doi.org/10.7555/JBR.36.20220003 |
_version_ | 1784723196227878912 |
---|---|
author | Ye, Wei Liu, Xiaoyu He, Ruiting Gou, Liming Lu, Ming Yang, Gang Wen, Jiaqi Wang, Xufei Liu, Fang Ma, Sujuan Qian, Weifeng Jia, Shaochang Ding, Tong Sun, Luan Gao, Wei |
author_facet | Ye, Wei Liu, Xiaoyu He, Ruiting Gou, Liming Lu, Ming Yang, Gang Wen, Jiaqi Wang, Xufei Liu, Fang Ma, Sujuan Qian, Weifeng Jia, Shaochang Ding, Tong Sun, Luan Gao, Wei |
author_sort | Ye, Wei |
collection | PubMed |
description | High-affinity antibodies are widely used in diagnostics and for the treatment of human diseases. However, most antibodies are isolated from semi-synthetic libraries by phage display and do not possess in vivo affinity maturation, which is triggered by antigen immunization. It is therefore necessary to engineer the affinity of these antibodies by way of in vitro assaying. In this study, we optimized the affinity of two human monoclonal antibodies which were isolated by phage display in a previous related study. For the 42A1 antibody, which targets the liver cancer antigen glypican-3, the variant T57H in the second complementarity-determining region of the heavy chain (CDR-H2) exhibited a 2.6-fold improvement in affinity, as well as enhanced cell-binding activity. For the I4A3 antibody to severe acute respiratory syndrome coronavirus 2, beneficial single mutations in CDR-H2 and CDR-H3 were randomly combined to select the best synergistic mutations. Among these, the mutation S53P-S98T improved binding affinity (about 3.7 fold) and the neutralizing activity (about 12 fold) compared to the parent antibody. Taken together, single mutations of key residues in antibody CDRs were enough to increase binding affinity with improved antibody functions. The mutagenic combination of key residues in different CDRs creates additive enhancements. Therefore, this study provides a safe and effective in vitro strategy for optimizing antibody affinity. |
format | Online Article Text |
id | pubmed-9179109 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Editorial Department of Journal of Biomedical Research |
record_format | MEDLINE/PubMed |
spelling | pubmed-91791092022-07-05 Improving antibody affinity through in vitro mutagenesis in complementarity determining regions Ye, Wei Liu, Xiaoyu He, Ruiting Gou, Liming Lu, Ming Yang, Gang Wen, Jiaqi Wang, Xufei Liu, Fang Ma, Sujuan Qian, Weifeng Jia, Shaochang Ding, Tong Sun, Luan Gao, Wei J Biomed Res Original Article High-affinity antibodies are widely used in diagnostics and for the treatment of human diseases. However, most antibodies are isolated from semi-synthetic libraries by phage display and do not possess in vivo affinity maturation, which is triggered by antigen immunization. It is therefore necessary to engineer the affinity of these antibodies by way of in vitro assaying. In this study, we optimized the affinity of two human monoclonal antibodies which were isolated by phage display in a previous related study. For the 42A1 antibody, which targets the liver cancer antigen glypican-3, the variant T57H in the second complementarity-determining region of the heavy chain (CDR-H2) exhibited a 2.6-fold improvement in affinity, as well as enhanced cell-binding activity. For the I4A3 antibody to severe acute respiratory syndrome coronavirus 2, beneficial single mutations in CDR-H2 and CDR-H3 were randomly combined to select the best synergistic mutations. Among these, the mutation S53P-S98T improved binding affinity (about 3.7 fold) and the neutralizing activity (about 12 fold) compared to the parent antibody. Taken together, single mutations of key residues in antibody CDRs were enough to increase binding affinity with improved antibody functions. The mutagenic combination of key residues in different CDRs creates additive enhancements. Therefore, this study provides a safe and effective in vitro strategy for optimizing antibody affinity. Editorial Department of Journal of Biomedical Research 2022-05 2022-03-28 /pmc/articles/PMC9179109/ /pubmed/35545451 http://dx.doi.org/10.7555/JBR.36.20220003 Text en © 2022 by the Journal of Biomedical Research. https://creativecommons.org/licenses/by/4.0/This is an open access article under the Creative Commons Attribution (CC BY 4.0) license, which permits others to distribute, remix, adapt and build upon this work, for commercial use, provided the original work is properly cited. |
spellingShingle | Original Article Ye, Wei Liu, Xiaoyu He, Ruiting Gou, Liming Lu, Ming Yang, Gang Wen, Jiaqi Wang, Xufei Liu, Fang Ma, Sujuan Qian, Weifeng Jia, Shaochang Ding, Tong Sun, Luan Gao, Wei Improving antibody affinity through in vitro mutagenesis in complementarity determining regions |
title | Improving antibody affinity through in vitro mutagenesis in complementarity determining regions
|
title_full | Improving antibody affinity through in vitro mutagenesis in complementarity determining regions
|
title_fullStr | Improving antibody affinity through in vitro mutagenesis in complementarity determining regions
|
title_full_unstemmed | Improving antibody affinity through in vitro mutagenesis in complementarity determining regions
|
title_short | Improving antibody affinity through in vitro mutagenesis in complementarity determining regions
|
title_sort | improving antibody affinity through in vitro mutagenesis in complementarity determining regions |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9179109/ https://www.ncbi.nlm.nih.gov/pubmed/35545451 http://dx.doi.org/10.7555/JBR.36.20220003 |
work_keys_str_mv | AT yewei improvingantibodyaffinitythroughinvitromutagenesisincomplementaritydeterminingregions AT liuxiaoyu improvingantibodyaffinitythroughinvitromutagenesisincomplementaritydeterminingregions AT heruiting improvingantibodyaffinitythroughinvitromutagenesisincomplementaritydeterminingregions AT gouliming improvingantibodyaffinitythroughinvitromutagenesisincomplementaritydeterminingregions AT luming improvingantibodyaffinitythroughinvitromutagenesisincomplementaritydeterminingregions AT yanggang improvingantibodyaffinitythroughinvitromutagenesisincomplementaritydeterminingregions AT wenjiaqi improvingantibodyaffinitythroughinvitromutagenesisincomplementaritydeterminingregions AT wangxufei improvingantibodyaffinitythroughinvitromutagenesisincomplementaritydeterminingregions AT liufang improvingantibodyaffinitythroughinvitromutagenesisincomplementaritydeterminingregions AT masujuan improvingantibodyaffinitythroughinvitromutagenesisincomplementaritydeterminingregions AT qianweifeng improvingantibodyaffinitythroughinvitromutagenesisincomplementaritydeterminingregions AT jiashaochang improvingantibodyaffinitythroughinvitromutagenesisincomplementaritydeterminingregions AT dingtong improvingantibodyaffinitythroughinvitromutagenesisincomplementaritydeterminingregions AT sunluan improvingantibodyaffinitythroughinvitromutagenesisincomplementaritydeterminingregions AT gaowei improvingantibodyaffinitythroughinvitromutagenesisincomplementaritydeterminingregions |