Construction of the single-chain Fv from 196-14 antibody toward ovarian cancer-associated antigen CA125.
The variable regions of heavy- and light-chains of mouse monoclonal antibody 196-14 toward ovarian cancer-associated antigen CA125 were linked with a peptide linker (GSTSGSGKSSEGKG) and a histidine tag was attached at the carboxyl terminal. This single-chain Fv (scFv) with a histidine tag was expressed in Escherichia coli as an inclusion body. The inclusion body was solubilized with guanidium chloride, followed by purification on nickel nitrilotriacetic acid agarose column and refolding into the active form. The scFv thus obtained bound to the Siso cells, which express CA125, and may recognize the same epitope as the parental 196-14 antibody IgG does.
Hashimoto
Y
Y
Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
Tanigawa
K
K
Nakashima
M
M
Sonoda
K
K
Ueda
T
T
Watanabe
T
T
Imoto
T
T
D000818:Animals; D000915:Antibody Affinity; D000951:Antigens, Neoplasm / Q000276:immunology; D001667:Binding, Competitive; D018394:CA-125 Antigen / Q000276:immunology*; D002478:Cells, Cultured; D005260:Female; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology*; D007074:Immunoglobulin G / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology; D051379:Mice; D010051:Ovarian Neoplasms / Q000175:diagnosis / Q000276:immunology*; D017510:Protein Folding; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
D000951:Antigens, Neoplasm; D018394:CA-125 Antigen; D007128:Immunoglobulin Fragments; D007074:Immunoglobulin G; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins