title: Rapid isolation of antigen-specific antibody-secreting cells using a chip-based immunospot array.
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.
authors.affiliation: Department of Immunology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama, Japan.
authors.forename: Aishun
authors.initials: A
authors.lastname: Jin
authors.forename: Tatsuhiko
authors.initials: T
authors.lastname: Ozawa
authors.forename: Kazuto
authors.initials: K
authors.lastname: Tajiri
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Obata
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kishi
authors.forename: Atsushi
authors.initials: A
authors.lastname: Muraguchi
mesh_terms: D000818:Animals; D000921:Antibody-Producing Cells / Q000166:cytology*; D058501:Enzyme-Linked Immunospot Assay / Q000379:methods*; D006801:Humans; D051379:Mice; D046888:Tissue Array Analysis / Q000379:methods*