Cell display library for gene cloning of variable regions of human antibodies to hepatitis B surface antigen.
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 VH and VK gene segments of the 4 clones isolated in this study belonged to VHIII and VKI 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.
Higuchi
K
K
Fundamental Research Laboratory of Life Science, Asahi Chemical Industry Co., Ltd., Shizuoka, Japan.
Araki
T
T
Matsuzaki
O
O
Sato
A
A
Kanno
K
K
Kitaguchi
N
N
Ito
H
H
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D015723:Gene Library; D005803:Genes, Immunoglobulin; D006510:Hepatitis B Antibodies / Q000235:genetics*; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D008969:Molecular Sequence Data; D014162:Transfection
D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D007135:Immunoglobulin Variable Region