Abstract

While achieving rapid developments in recent years, bispecific antibodies are still difficult to design and manufacture, due to mispair of both heavy and light chains. Here we report a novel technology to make bispecific molecules. The knob-into-hole method was used to pair two distinct heavy chains as a heterodimer. IgG4 S228P CH1-CL interface was then partially replaced by T-cell receptor α/β constant domain to increase the efficiency of cognate heavy and light chain pairing. Following expression and purification, the bispecific antibody interface exchange was confirmed by Western blotting and LC–MS/MS. To ensure its validity, we combined a monovalent bispecific antibody against PD-1 (sequence from Pembrolizumab) and LAG3 (sequence from Relatlimab). The results showed that the molecule could be assembled correctly at a ratio of 95% in cells. In vitro functional assay demonstrated that the purified bispecific antibody exhibits an enhanced agonist activity compared to that of the parental antibodies. Low immunogenicity was predicted by an open-access software and ADA test.

Details

Title
A new approach to produce IgG4-like bispecific antibodies
Author
Zhao Caizhi 1 ; Zhang, Wei 2 ; Gong Guihua 2 ; Xie Liping 2 ; Ming-Wei, Wang 3 ; Hu Youjia 2 

 Fudan University, School of Pharmacy, Shanghai, China (GRID:grid.8547.e) (ISNI:0000 0001 0125 2443); China State Institute of Pharmaceutical Industry, Shanghai, China (GRID:grid.419098.d) (ISNI:0000 0004 0632 441X) 
 China State Institute of Pharmaceutical Industry, Shanghai, China (GRID:grid.419098.d) (ISNI:0000 0004 0632 441X) 
 Fudan University, School of Pharmacy, Shanghai, China (GRID:grid.8547.e) (ISNI:0000 0001 0125 2443); The National Center for Drug Screening, Shanghai, China (GRID:grid.410611.3) (ISNI:0000 0004 7699 6713) 
Publication year
2021
Publication date
2021
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
2574549768
Copyright
© The Author(s) 2021. corrected publication 2022. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.