Use of detergent-based buffers allows detection forms in an immunoassay format

Inhibin ELISAs are used in monitoring aspects of reproductive function, however these assays are based on the measurements of the mature 30 kDa inhibin forms and not precursor forms. In conventional ELISA formats, the 105 kDa unprocessed 'Pro-inhibin' forms are immunologically inactive, but the immunoactivity can be recovered in the presence of detergents. The immunoactivity of Pro-inhibin forms was assessed in the presence of a range of detergents utilising antibodies to the alpha-, beta(A)- and beta(B)-subunits of inhibin. In contrast to mature forms, unprocessed inhibin forms showed a 10-40 fold increase in inhibin A and total inhibin immunoactivities under optimised detergent (0.5% SDS/2% Triton X-100) conditions. The suppressed immunoactivity of the Pro-inhibin forms in these immunoassays was attributed to steric hindrance by the respective pp,beta(A)- and alpha-subunit prodomains. This study details a detergent-based immunoassay that allows detection of previously undetectable precursor inhibin forms. (C) 2013 Elsevier Ireland Ltd. All rights reserved.