The new solid-phase immunoassay technique has been applied for antidigoxin monoclonal antibodies (Mabs) detection. At the first assay step, antidigoxin Mabs (IgG1, K(aff) = 9.2·109 M-1) bound with specially prepared immunosorbent: microtiter plates with adsorbed digoxin-human serum albumin conjugate, in which free amino groups were blocked by glutaraldehyde (the microtiter plates with immobilized conjugate were treated for 3 h by 2.5% glutaraldehyde at 45°C, this had no significant effect on the immunological properties of immunosorbents). After antigen-antibody reaction and solid phase separation, free amino groups were located only at the antidigoxin Mabs bound to GSI immunosorbent. At the second assay step, N-hydroxysuccinimidebiotin ester solution was added to bind with the free amino groups. After separation at the third assay step, biotin labels were detected by the streptavidin-peroxidase conjugate. The method does not require second labeled antibodies and may be used for anti-hapten screening.
|Number of pages||9|
|Publication status||Published - 1995|
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