利用 Beacon 平台从免疫小鼠体内分离抗原特异性单克隆抗体的方案。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2024-12-20 Epub Date: 2024-10-12 DOI:10.1016/j.xpro.2024.103389

Meilan Fu, Xiangwei Wang, Shubing Tang, Shimeng Bai, Longfei Ding, Yangyang Hu, Kangli Cao, Tianhan Yang, Chen Zhao, Xiaoyan Zhang, Jianqing Xu

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引用次数: 0

摘要

在 Beacon 平台问世之前，抗原特异性血浆 B 细胞的分离一直是个难题。利用光分选技术，Beacon 可以在芯片上对血浆 B 细胞进行高通量筛选，分选出具有所需抗原特异性的单细胞。在这里，我们介绍一种利用 Beacon 从免疫小鼠体内分离抗原特异性血浆 B 细胞、对编码的 B 细胞受体（BCR）进行测序、克隆和表达由此产生的抗体的方案。该方案可轻松扩展到人类样本。

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Protocol for isolating antigen-specific monoclonal antibodies from immunized mice utilizing the Beacon platform.

Isolation of antigen-specific plasma B cells had been challenging until the recent arrival of the Beacon platform. Leveraging light-sorting technology, Beacon can perform high-throughput screening of plasma B cells on a chip to sort single cells with the desired antigen specificity. Here, we present a protocol for isolating antigen-specific plasma B cells from immunized mice using Beacon, sequencing the encoded B cell receptors (BCRs), and cloning and expressing the resulting antibodies. This protocol can easily be extended to human samples.

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