Massively parallel single-cell B-cell receptor sequencing enables rapid discovery of diverse antigen-reactive antibodies

作者：Leonard D. Goldstein, Ying-Jiun J. Chen, Jia Wu, Subhra Chaudhuri, Yi-Chun Hsiao, Kellen Schneider, Kam Hon Hoi, Zhonghua Lin, Steve Guerrero, Bijay S. Jaiswal, Jeremy Stinson, Aju Antony, Kanika Bajaj Pahuja, Dhaya Seshasayee, Zora Modrušan, Isidro Hötzel, Somasekar Seshagiri · 发表于：Communications Biology · 年份：2019 · DOI：10.1038/s42003-019-0551-y · 被引用次数：180 · 研究领域：Monoclonal and Polyclonal Antibodies Research、T-cell and B-cell Immunology、Viral Infectious Diseases and Gene Expression in Insects

Abstract Obtaining full-length antibody heavy- and light-chain variable regions from individual B cells at scale remains a challenging problem. Here we use high-throughput single-cell B-cell receptor sequencing (scBCR-seq) to obtain accurately paired full-length variable regions in a massively parallel fashion. We sequenced more than 250,000 B cells from rat, mouse and human repertoires to characterize their lineages and expansion. In addition, we immunized rats with chicken ovalbumin and profiled antigen-reactive B cells from lymph nodes of immunized animals. The scBCR-seq data recovered 81% ( n = 56/69) of B-cell lineages identified from hybridomas generated from the same set of B cells subjected to scBCR-seq. Importantly, scBCR-seq identified an additional 710 candidate lineages not recovered as hybridomas. We synthesized, expressed and tested 93 clones from the identified lineages and found that 99% ( n = 92/93) of the clones were antigen-reactive. Our results establish scBCR-seq as a powerful tool for antibody discovery.