Massively parallel single-cell B-cell receptor sequencing enables rapid discovery of diverse antigen-reactive antibodies
Massively parallel single-cell B-cell receptor sequencing enables rapid discovery of diverse antigen-reactive antibodies
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DOI：
10.1038/s42003-019-0551-y
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发表时间：
2019-08-09
影响因子：
5.9
通讯作者：
Seshagiri, Somasekar
中科院分区：
文献类型：
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作者：
Goldstein, Leonard D.;Chen, Ying-Jiun J.;Seshagiri, Somasekar
从单个B细胞大规模获得全长抗体重链和轻链可变区仍然是一个具有挑战性的问题。在这里，我们使用高通量单细胞B细胞受体测序（scBCR-seq）以大规模平行的方式获得准确配对的全长可变区。我们对来自大鼠、小鼠和人类的25万多个B细胞进行了测序，以表征其谱系和扩增。此外，我们用鸡卵清蛋白免疫大鼠，并从免疫动物的淋巴结中分析抗原反应性B细胞。scBCR-seq数据回收了81%（n = 56/69）的从杂交瘤鉴定的B细胞谱系，所述杂交瘤产生自经受scBCR-seq的同一组B细胞。重要的是，scBCR-seq鉴定了另外710个未作为杂交瘤回收的候选谱系。我们合成、表达并测试了来自鉴定的谱系的93个克隆，发现99%（n = 92/93）的克隆是抗原反应性的。我们的研究结果确立了scBCR-seq作为抗体发现的有力工具。
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.