Cell display library for gene cloning of variable regions of human antibodies to hepatitis B surface antigen

Kazuo Higuchi, Takeyoshi Araki, Osamu Matsuzaki, Akiko Sato, Kimiyoshi Kanno, Nobuya Kitaguchi, Hirataka Ito

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

A novel cell display system was developed for cloning the variable region (V) genes of antigen-specific human antibodies. The system is based on an antibody library displayed on the surface of COS cells, using a plasmid vector designed to direct expression of membrane-bound antibodies. COS cells expressing antigen-specific antibodies were separated using a flow cytometer for their binding to a fluorescent dye-labeled antigen. To test the performance of this system, we cloned V genes of 4 antibodies directed against hepatitis B surface antigen (HBsAg) from a library prepared from peripheral blood lymphocytes of a vaccinated donor. These membrane-bound anti-HBsAg antibodies were easily converted to soluble forms, all of which showed a size similar to human serum IgG in SDS-PAGE and the same specific binding to HBsAg as membrane-bound forms in ELISA. All V(H) and Y(κ) gene segments of the 4 clones isolated in this study belonged to V(H)III and V(κ)I subgroups, respectively. These findings demonstrate the potential and selection capabilities of our cell display system for cloning the V genes of antigen-specific human antibodies.

Original languageEnglish
Pages (from-to)193-204
Number of pages12
JournalJournal of Immunological Methods
Volume202
Issue number2
DOIs
Publication statusPublished - 28-03-1997

Fingerprint

Hepatitis B Surface Antigens
Gene Library
Organism Cloning
Antibodies
Antigens
COS Cells
Genes
Membranes
Hepatitis B Antibodies
Fluorescent Dyes
Libraries
Polyacrylamide Gel Electrophoresis
Plasmids
Clone Cells
Immunoglobulin G
Enzyme-Linked Immunosorbent Assay
Lymphocytes
Serum

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

Cite this

Higuchi, Kazuo ; Araki, Takeyoshi ; Matsuzaki, Osamu ; Sato, Akiko ; Kanno, Kimiyoshi ; Kitaguchi, Nobuya ; Ito, Hirataka. / Cell display library for gene cloning of variable regions of human antibodies to hepatitis B surface antigen. In: Journal of Immunological Methods. 1997 ; Vol. 202, No. 2. pp. 193-204.
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Cell display library for gene cloning of variable regions of human antibodies to hepatitis B surface antigen. / Higuchi, Kazuo; Araki, Takeyoshi; Matsuzaki, Osamu; Sato, Akiko; Kanno, Kimiyoshi; Kitaguchi, Nobuya; Ito, Hirataka.

In: Journal of Immunological Methods, Vol. 202, No. 2, 28.03.1997, p. 193-204.

Research output: Contribution to journalArticle

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