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利用Beacon平台从免疫小鼠中分离抗原特异性单克隆抗体的方案。

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

作者信息

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

机构信息

Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, China.

Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, China; Department of Investigational New Drug, Shanghai Reinovax Biologics Co., Ltd., Shanghai, China.

出版信息

STAR Protoc. 2024 Dec 20;5(4):103389. doi: 10.1016/j.xpro.2024.103389. Epub 2024 Oct 12.

Abstract

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.

摘要

在Beacon平台出现之前,分离抗原特异性浆B细胞一直具有挑战性。利用光分选技术,Beacon可以在芯片上对浆B细胞进行高通量筛选,以分选具有所需抗原特异性的单细胞。在此,我们展示了一种使用Beacon从免疫小鼠中分离抗原特异性浆B细胞、对编码的B细胞受体(BCR)进行测序以及克隆和表达所得抗体的方案。该方案可以很容易地扩展到人类样本。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/857e/11735987/3dd491a45c2c/fx1.jpg

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