Preparation and application of anti-idiotypic antibody against anti-gibberellin A4 antibody.
A monoclonal anti-idiotypic antibody was raised against anti-gibberellin A4 (GA4) antibody, which recognizes biologically active gibberellins such as GA1 and GA4 specifically. Amino acid sequences of variable regions of both anti-GA4 and anti-idiotypic antibodies were analyzed. By using the property of the anti-idiotypic antibody to compete with GA1/4 in binding to the anti-GA4 antibody, we successfully applied the anti-idiotypic antibody to ELISA as a tracer for measuring GA1/4. The single-chain Fv (scFv) gene of the anti-idiotypic antibody was constructed, and scFv expressed in E. coli showed binding activity to anti-GA4 antibody. These results suggest the possible application of anti-idiotypic antibody as a handy and stable source of an enzymatic tracer for ELISA by production of fusion protein of the scFv and an appropriate enzyme.
Suzuki
Y
Y
Department of Applied Biological Chemistry, University of Tokyo, Japan. ayoshi@hongo.ecc.u-tokyo.ac.jp
Shimada
N
N
Niwa
R
R
Yokoi
M
M
Nakajima
M
M
Murofushi
N
N
Yamaguchi
I
I
D000595:Amino Acid Sequence; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000096:biosynthesis / Q000302:isolation & purification*; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000302:isolation & purification*; D001483:Base Sequence; D002459:Cell Fusion; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000378:metabolism; D005260:Female; D005875:Gibberellins / Q000276:immunology*; D006825:Hybridomas; D007074:Immunoglobulin G / Q000276:immunology; D007536:Isomerism; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D011863:Radioimmunoassay
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D005875:Gibberellins; D007074:Immunoglobulin G