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Group-selective immunoassay

S A Yazynin1, S M Deyev

  • 1Department of Molecular Genetic Immunology, V.A. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow.

Immunology Letters
|July 1, 1994
PubMed

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 x 10(9) 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.

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