A rapid and efficient single-cell manipulation method for screening antigen-specific antibody-secreting cells from human peripheral blood
A rapid and efficient single-cell manipulation method for screening antigen-specific antibody-secreting cells from human peripheral blood
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DOI：
10.1038/nm.1966
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发表时间：
2009-09-01
期刊：
影响因子：
82.9
通讯作者：
Muraguchi, Atsushi
中科院分区：
文献类型：
--
作者：
Jin, Aishun;Ozawa, Tatsuhiko;Muraguchi, Atsushi
抗原特异性人单克隆抗体（mAb）是治疗剂的关键候选物。然而，一个合适的筛选系统的抗原特异性抗体分泌细胞（ASC）的可用性是有限的，在人类。在这里，我们提出了一种使用微孔阵列芯片检测单个ASC的独特方法，该方法能够在单细胞基础上分析活细胞，并提供快速，高效和高通量（高达234，000个单个细胞）的系统用于识别和回收目标ASC。我们应用该系统从人外周血淋巴细胞中检测和回收了针对B型肝炎病毒和流感病毒的ASCs，并在一周内产生了具有病毒中和活性的人mAb。此外，我们表明，该系统是有用的检测ASCs的多种抗原，以及选择ASCs分泌高亲和力抗体的芯片上。我们的方法可以为个体患者的治疗性抗体的产生开辟道路。
Antigen-specific human monoclonal antibodies (mAbs) are key candidates for therapeutic agents. However, the availability of a suitable screening system for antigen-specific antibody-secreting cells (ASCs) is limited in humans. Here we present a unique method for detecting individual ASCs using microwell array chips, which enables the analysis of live cells on a single-cell basis and offers a rapid, efficient and high-throughput (up to 234,000 individual cells) system for identifying and recovering objective ASCs. We applied the system to detect and retrieve ASCs for hepatitis B virus and influenza viruses from human peripheral blood lymphocytes and produced human mAbs with virus-neutralizing activities within a week. Furthermore, we show that the system is useful for detecting ASCs for multiple antigens as well as for selection of ASCs secreting high-affinity antibodies on a chip. Our method can open the way for the generation of therapeutic antibodies for individual patients.