Group-selective immunoassay

Sergey A. Yazynin, Sergey M. Deyev

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

A new solid-phase immunoassay technique has been applied for anti-digoxin monoclonal antibodies (Mabs) detection. At the first assay step, anti-digoxin Mabs (IgG1, Kaff = 9.2 × 109 M-1) were bound to a specially prepared immunosorbent, the microtiter plates coated with digoxin-human serum albumin conjugate (Dig-HSA), in which free amino groups were protected by a glutaraldehyde cross-linking modification. The modification did not essentially influence the antibody-binding capacity of the immunosorbent. After antigen-antibody reaction, free amino groups were located only on the anti-digoxin Mabs, bound to chemically modified immunosorbent. At the second assay step, free amino groups of anti-digoxin Mabs were biotinylated by N-hydroxysuccinimide-biotin ester. Then the biotin residues were detected by the streptavidin-peroxidase conjugate. The method does not require second labeled antibodies and may be used for anti-hapten hybridoma screening.

Original languageEnglish
Pages (from-to)235-239
Number of pages5
JournalImmunology Letters
Volume41
Issue number2-3
DOIs
Publication statusPublished - 1994
Externally publishedYes

Keywords

  • Digoxin
  • Group-selective immunoassay
  • Monoclonal antibody
  • Streptavidin-biotin system

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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