J Korean Med Sci.  1996 Oct;11(5):390-396. 10.3346/jkms.1996.11.5.390.

Monoclonal antibody production and characterization for the measurement of plasma high density lipoprotein

• Lee SH
• Cho MH

• KMID: 644996
• DOI: http://doi.org/10.3346/jkms.1996.11.5.390

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.

Keyword

ELISA; immunoprecipitation; High density lipoprotein; Western blotting; Dissociation constant (Kd)

MeSH Terms

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