Rapid isolation of antigen-specific antibody-secreting cells using a chip-based immunospot array |
| |
Authors: | Jin Aishun Ozawa Tatsuhiko Tajiri Kazuto Obata Tsutomu Kishi Hiroyuki Muraguchi Atsushi |
| |
Institution: | Department of Immunology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama, Japan. |
| |
Abstract: | 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. |
| |
Keywords: | |
本文献已被 PubMed 等数据库收录! |
|