Jin Aishun, Ozawa Tatsuhiko, Tajiri Kazuto, Obata Tsutomu, Kondo Sachiko, Kinoshita Koshi, Kadowaki Shinichi, Takahashi Kazuo, Sugiyama Toshiro, Kishi Hiroyuki, Muraguchi Atsushi
Department of Immunology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama, Japan.
Nat Med. 2009 Sep;15(9):1088-92. doi: 10.1038/nm.1966. Epub 2009 Aug 16.
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.
抗原特异性人单克隆抗体（mAb）是治疗药物的关键候选物。然而，人类中用于筛选抗原特异性抗体分泌细胞（ASC）的合适筛选系统有限。在此，我们展示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，并提供了一个快速、高效且高通量（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检索针对乙型肝炎病毒和流感病毒的ASC，并在一周内产生了具有病毒中和活性的人mAb。此外，我们表明该系统可用于检测多种抗原的ASC，以及在芯片上选择分泌高亲和力抗体的ASC。我们的方法可为为个体患者生成治疗性抗体开辟道路。
