Radial-immunodiffusion assay of human apolipoprotein A-I with use of two monoclonal antibodies combined

S. Marcovina, G. Di Cola, A. L. Catapano

Research output: Contribution to journalArticlepeer-review

Abstract

We produced and characterized several monoclonal antibodies directed toward human plasma apolipoprotein A-I. Two of them, A-I-12 and A-I-57, individually precipitated purified or native high-density lipoprotein in agarose gel by double immunodiffusion. Because radial immunodiffusion performed with a single monoclonal antibody gave faint and diffuse rings of precipitation, we developed and optimized working conditions for using these two monoclonal antibodies combined to determine apolipoprotein A-I in human plasma. This combination gave easy-to-measure, clear, sharp rings, and linear and parallel standard curves for HDL3 (the primary standard) and a reference serum (the secondary standard). Moreover, no pretreatment of samples with dissociating agents or detergents is necessary. The assay was complete after overnight incubation, as compared with two to three days when polyclonal antisera were used. Apolipoprotein A-I concentrations as measured in 128 normolipidemic subjects and in 72 patients with various lipid disorders by the radial immunodiffusion technique with monoclonal antibodies (x) compared well (r = 0.882; y = 1.029x - 0.036) with those measured by radial immunodiffusion with polyclonal antisera (y).

Original languageEnglish
Pages (from-to)2155-2159
Number of pages5
JournalClinical Chemistry
Volume32
Issue number12
Publication statusPublished - 1986

ASJC Scopus subject areas

  • Clinical Biochemistry

Fingerprint Dive into the research topics of 'Radial-immunodiffusion assay of human apolipoprotein A-I with use of two monoclonal antibodies combined'. Together they form a unique fingerprint.

Cite this