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 language | English |
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Pages (from-to) | 235-239 |
Number of pages | 5 |
Journal | Immunology Letters |
Volume | 41 |
Issue number | 2-3 |
DOIs | |
Publication status | Published - 1994 |
Externally published | Yes |
Keywords
- Digoxin
- Group-selective immunoassay
- Monoclonal antibody
- Streptavidin-biotin system
ASJC Scopus subject areas
- Immunology and Allergy
- Immunology