Monoclonal antibody production and characterization for the measurement of plasma high density lipoprotein.
10.3346/jkms.1996.11.5.390
- Author:
Sang Han LEE
1
;
Man Hee CHO
Author Information
1. Department of Biochemistry, College of Medicine, Soonchunhyang University, Cheon-An, Korea.
- Publication Type:Original Article
- Keywords:
ELISA;
immunoprecipitation;
High density lipoprotein;
Western blotting;
Dissociation constant (Kd)
- MeSH:
Animal;
Antibodies, Monoclonal/*immunology/isolation & purification;
Antibody Affinity;
Apolipoprotein A-I/*immunology;
Blotting, Western;
Hybridomas;
Lipoproteins, HDL/immunology;
Mice;
Mice, Inbred BALB C;
Tumor Cells, Cultured
- From:Journal of Korean Medical Science
1996;11(5):390-396
- CountryRepublic of Korea
- Language:English
-
Abstract:
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.
