Improving antibody affinity through in vitro mutagenesis in complementarity determining regions

作     者：Wei Ye Xiaoyu Liu Ruiting He Liming Gou Ming Lu Gang Yang Jiaqi Wen Xufei Wang Fang Liu Sujuan Ma Weifeng Qian Shaochang Jia Tong Ding Luan Sun Wei Gao 

作者机构：Jiangsu Key Lab of Cancer BiomarkersPrevention and TreatmentCollaborative Innovation Center for Personalized Cancer MedicineKey Laboratory of Human Functional Genomics of Jiangsu ProvinceNational Health Commission Key Laboratory of Antibody TechniquesSchool of Basic Medical SciencesNanjing Medical UniversityNanjingJiangsu 211166China The Affiliated Suzhou Hospital of Nanjing Medical UniversitySuzhou Municipal HospitalGusu SchoolNanjing Medical UniversitySuzhouJiangsu 215001China Department of BiotherapyNanjing Jinling HospitalNanjingJiangsu 210002China. 

出 版 物：《The Journal of Biomedical Research》 (生物医学研究杂志（英文版）)

年 卷 期：2022年第36卷第3期

页      面：155-166页

核心收录：

学科分类：1001[医学-基础医学(可授医学、理学学位)] 100102[医学-免疫学] 10[医学] 

基　　金：supported by the National Natural Science Foundation of China (Grant No. 81972284) 

主　　题：antibody engineering phage display affinity maturation 

摘      要：High-affinity antibodies are widely used in diagnostics and for the treatment of human ***,most antibodies are isolated from semi-synthetic libraries by phage display and do not possess in vivo affinity maturation,which is triggered by antigen *** is therefore necessary to engineer the affinity of these antibodies by way of in vitro *** this study,we optimized the affinity of two human monoclonal antibodies which were isolated by phage display in a previous related *** 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 *** 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 *** these,the mutation S53P-S98T improved binding affinity(about 3.7 fold)and the neutralizing activity(about 12 fold)compared to the parent *** together,single mutations of key residues in antibody CDRs were enough to increase binding affinity with improved antibody *** mutagenic combination of key residues in different CDRs creates additive ***,this study provides a safe and effective in vitro strategy for optimizing antibody affinity.