title: A rapid and efficient single-cell manipulation method for screening antigen-specific antibody-secreting cells from human peripheral blood.
abstract: 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.
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: Sachiko
authors.initials: S
authors.lastname: Kondo
authors.forename: Koshi
authors.initials: K
authors.lastname: Kinoshita
authors.forename: Shinichi
authors.initials: S
authors.lastname: Kadowaki
authors.forename: Kazuo
authors.initials: K
authors.lastname: Takahashi
authors.forename: Toshiro
authors.initials: T
authors.lastname: Sugiyama
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kishi
authors.forename: Atsushi
authors.initials: A
authors.lastname: Muraguchi
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000627:therapeutic use; D000914:Antibodies, Viral / Q000096:biosynthesis; D000918:Antibody Specificity; D000921:Antibody-Producing Cells / Q000166:cytology* / Q000276:immunology*; D002469:Cell Separation; D002645:Chickens; D006510:Hepatitis B Antibodies / Q000096:biosynthesis; D006801:Humans; D006825:Hybridomas / Q000166:cytology / Q000276:immunology; D066298:In Vitro Techniques; D008214:Lymphocytes / Q000166:cytology / Q000276:immunology; D051379:Mice; D046228:Microarray Analysis / Q000379:methods*; D009113:Muramidase / Q000276:immunology; D009500:Neutralization Tests; D009975:Orthomyxoviridae / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D006510:Hepatitis B Antibodies; D009113:Muramidase
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