Monoclonal antibody production and characterization for the measurement of plasma high density lipoprotein.
We prepared two monoclonal antibodies-A-I30 and A-I4 to HDL apo A-I-with the ultimate goals of expressing and overproducing the valuable immunodiagnostic single chain Fv by phage display libraries in E, coli. Monoclonal antibodies were produced by immunizing mice with apolipoprotein A-I, and purified from ascitic fluid by affinity chromatography on a Protein A Sepharose CL-4B column. The specificity of A-I30 and A-I4 was confirmed by ELISA and Western blotting. The dissociation constants(Kd) of antigen-antibody complex obtained by enzyme linked immunosorbent assay(ELISA) method were 2.25 x 10(-8) M for A-I30 and 2.15 x 10(-8) M for A-I4. The experimental values of Kd are shown to be close to those obtained by immunoprecipitation of the radiolabeled antigen. From the above results, it was shown that A-I30 and A-I4 could be provided as excellent reagents for the determination of plasma HDL concentrations in clinical specimens using ELISA.
Lee
S H
SH
Department of Biochemistry, College of Medicine, Soonchunhyang University, Cheon-An, Korea.
Cho
M H
MH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000915:Antibody Affinity; D016632:Apolipoprotein A-I / Q000276:immunology*; D015153:Blotting, Western; D006825:Hybridomas; D008075:Lipoproteins, HDL / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D016632:Apolipoprotein A-I; D008075:Lipoproteins, HDL