A rapid and efficient single-cell manipulation method for screening antigen-specific antibody-secreting cells from human peripheral blood.
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.
Jin
Aishun
A
Department of Immunology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama, Japan.
Ozawa
Tatsuhiko
T
Tajiri
Kazuto
K
Obata
Tsutomu
T
Kondo
Sachiko
S
Kinoshita
Koshi
K
Kadowaki
Shinichi
S
Takahashi
Kazuo
K
Sugiyama
Toshiro
T
Kishi
Hiroyuki
H
Muraguchi
Atsushi
A
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
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D006510:Hepatitis B Antibodies; D009113:Muramidase
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