Rapid isolation of antigen-specific antibody-secreting cells using a chip-based immunospot array
Rapid isolation of antigen-specific antibody-secreting cells using a chip-based immunospot array
复制标题
使用基于芯片的免疫点阵列快速分离抗原特异性抗体分泌细胞
DOI：
10.1038/nprot.2011.322
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发表时间：
2011-05-01
期刊：
影响因子：
14.8
通讯作者：
Muraguchi, Atsushi
中科院分区：
文献类型：
--
作者：
Jin, Aishun;Ozawa, Tatsuhiko;Muraguchi, Atsushi
在这里，我们报告了一种使用微孔阵列芯片分离抗原特异性抗体分泌细胞（ASCs）的新方法，该方法提供了一种快速，高效和高通量（高达234，000个单个细胞）的系统，用于检测和检索在单细胞基础上分泌感兴趣抗体的细胞。我们在微孔阵列芯片上排列了大量含有人外周血ASC的淋巴细胞，并用分泌的抗体检测斑点。该方案可以在不到7小时内完成，包括3小时的细胞培养。本文提出的方法不仅具有与酶联免疫斑点（ELISPOT）相当的高灵敏度和特异性，而且克服了ELISPOT在回收可用于产生抗原特异性人单克隆抗体的ASCs方面的局限性。该方法还可用于检测分泌抗体以外的分子的细胞，如细胞因子，为细胞分析和临床诊断提供了工具。
Here we report a new method for isolating antigen-specific antibody-secreting cells (ASCs) using a microwell array chip, which offers a rapid, efficient and high-throughput (up to 234,000 individual cells) system for the detection and retrieval of cells that secrete antibodies of interest on a single-cell basis. We arrayed a large population of lymphoid cells containing ASCs from human peripheral blood on microwell array chips and detected spots with secreted antibodies. This protocol can be completed in less than 7 h, including 3 h of cell culture. The method presented here not only has high sensitivity and specificity comparable with enzyme-linked immunospot (ELISPOT) but it also overcomes the limitations of ELISPOT in recovering ASCs that can be used to produce antigen-specific human monoclonal antibodies. This method can also be used to detect cells secreting molecules other than antibodies, such as cytokines, and it provides a tool for cell analysis and clinical diagnosis.