Epitope-directed antibody selection by site-specific photocrosslinking
Longxin Chen, Chaoyang Zhu, Hui Guo1, Runting Li, Limeng Zhang, Zhenzhen Xing, Yue Song, Zihan Zhang, Fuping Wang, Xiaofeng Liu, Yuhan Zhang, Runlin Z. Ma and Feng Wang
Abstract
Currently, there are no methods available offering solutions to select and identify antibodies binding to a specific conformational epitope of an antigen. Here, we developed a method to allow epitope-directed antibody selection from a phage display library by photocrosslinking bound antibodies to a site that specifically incorporates a noncanonical amino acid, p-benzoyl-L-phenylalanine (pBpa), on the target antigen epitope. By one or two rounds of panning against antibody phage display libraries, those hits that covalently bind to the proximity site of pBpa on specific epitopes of target antigens after ultraviolet irradiation are enriched and selected. This method was applied to specific epitopes on human interleukin-1β and complement 5a. In both cases, more than one-third of hits identified bind to the target epitopes, demonstrating the feasibility and versatility of this method.
附件下载：
Epitope-directed antibody selection by site-specific photocrosslinking, Sci Adv, 1 Apr 2020
Science Advances, 1 April, 2020, DOI：https://doi.org/10.1126/sciadv.aaz7825
Epitope-directed antibody selection by site-specific photocrosslinking
Longxin Chen, Chaoyang Zhu, Hui Guo1, Runting Li, Limeng Zhang, Zhenzhen Xing, Yue Song, Zihan Zhang, Fuping Wang, Xiaofeng Liu, Yuhan Zhang, Runlin Z. Ma and Feng Wang
Abstract
Currently, there are no methods available offering solutions to select and identify antibodies binding to a specific conformational epitope of an antigen. Here, we developed a method to allow epitope-directed antibody selection from a phage display library by photocrosslinking bound antibodies to a site that specifically incorporates a noncanonical amino acid, p-benzoyl-L-phenylalanine (pBpa), on the target antigen epitope. By one or two rounds of panning against antibody phage display libraries, those hits that covalently bind to the proximity site of pBpa on specific epitopes of target antigens after ultraviolet irradiation are enriched and selected. This method was applied to specific epitopes on human interleukin-1β and complement 5a. In both cases, more than one-third of hits identified bind to the target epitopes, demonstrating the feasibility and versatility of this method.
文章链接：https://advances.sciencemag.org/content/6/14/eaaz7825