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

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Massively parallel single-cell B-cell receptor sequencing enables rapid discovery of diverse antigen-reactive antibodies.
Communications Biology ( IF 5.2 ) Pub Date : 2019-08-09 , DOI: 10.1038/s42003-019-0551-y
Leonard D Goldstein _{1,

2} , Ying-Jiun J Chen ₁ , Jia Wu ₃ , Subhra Chaudhuri ₁ , Yi-Chun Hsiao ₃ , Kellen Schneider ₃ , Kam Hon Hoi _{2,

3} , Zhonghua Lin ₃ , Steve Guerrero ₂ , Bijay S Jaiswal ₁ , Jeremy Stinson ₁ , Aju Antony ₄ , Kanika Bajaj Pahuja ₁ , Dhaya Seshasayee ₃ , Zora Modrusan ₁ , Isidro Hötzel ₃ , Somasekar Seshagiri _{1,

5}

Affiliation

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.

中文翻译：

大规模并行单细胞 B 细胞受体测序能够快速发现多种抗原反应性抗体。

从单个 B 细胞中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) 以大规模并行方式获得精确配对的全长可变区。我们对来自大鼠、小鼠和人类的 250,000 多个 B 细胞进行了测序，以表征它们的谱系和扩增。此外，我们用鸡卵清蛋白对大鼠进行了免疫接种，并分析了来自免疫动物淋巴结的抗原反应性 B 细胞。 scBCR-seq 数据恢复了从接受 scBCR-seq 的同一组 B 细胞产生的杂交瘤中鉴定出的 B 细胞谱系的 81% (n = 56/69)。重要的是，scBCR-seq 鉴定了另外 710 个未恢复为杂交瘤的候选谱系。我们合成、表达并测试了来自已鉴定谱系的 93 个克隆，发现 99% (n = 92/93) 的克隆具有抗原反应性。我们的结果确立了 scBCR-seq 作为抗体发现的强大工具。

更新日期：2019-08-09

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