{"title":"Protocol for isolating antigen-specific monoclonal antibodies from immunized mice utilizing the Beacon platform.","authors":"Meilan Fu, Xiangwei Wang, Shubing Tang, Shimeng Bai, Longfei Ding, Yangyang Hu, Kangli Cao, Tianhan Yang, Chen Zhao, Xiaoyan Zhang, Jianqing Xu","doi":"10.1016/j.xpro.2024.103389","DOIUrl":null,"url":null,"abstract":"<p><p>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.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"5 4","pages":"103389"},"PeriodicalIF":1.3000,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"STAR Protocols","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1016/j.xpro.2024.103389","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/10/12 0:00:00","PubModel":"Epub","JCR":"Q4","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
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.
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。