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

被引：0
作者：
Fu, Meilan [1 ]
Wang, Xiangwei [1 ]
Tang, Shubing [1 ,3 ]
Bai, Shimeng [1 ,4 ]
Ding, Longfei [1 ]
Hu, Yangyang [1 ]
Cao, Kangli [2 ]
Yang, Tianhan [1 ]
Zhao, Chen [1 ]
Zhang, Xiaoyan [1 ,2 ,5 ,6 ]
Xu, Jianqing [1 ,2 ,5 ,6 ]
机构：
[1] Fudan Univ, Shanghai Publ Hlth Clin Ctr, Shanghai 201508, Peoples R China
[2] Fudan Univ, Zhongshan Hosp, Inst Biomed Sci, Shanghai, Peoples R China
[3] Shanghai Reinovax Biol Co Ltd, Dept Invest New Drug, Shanghai, Peoples R China
[4] Southern Univ Sci & Technol, Shenzhen Peoples Hosp 3, Biotherapeut Ctr, Natl Clin Res Ctr Infect Dis,Hosp Affiliated 2,Sch, Shenzhen, Guangdong, Peoples R China
[5] Xiamen Key Lab Biotherapy, Xiammen, Peoples R China
[6] Fudan Univ, Zhongshan Hosp Xiamen, Clin Ctr Biotherapy, Xiamen, Peoples R China
来源：
STAR PROTOCOLS | 2024年 / 5卷 / 04期
关键词：
Antibody; Cell isolation; Protein expression and purification; Sequencing; Single Cell;
D O I：
10.1016/j.xpro.2024.103389
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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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页数：26
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