Preparation of an ecdysone immunogen for radioimmunoassay work.
A highly improved procedure for the preparation of ecdysone-protein conjugates for immunological work is reported. Bovine thyroglobulin is succinylated and the succinylated protein is coupled to beta-ecdysone with 1-ethyl-3-(-dimethylaminopropyl)-carbodiimide in the presence of the acylation catalyst 4-dimethylaminopyridine, The antiserum obtained using this immunogen provides a radioimmunoassay sensitive to 25 pmol of beta-ecdysone. The anti-ecdysone antibody cross-reacts with alpha-ecdysone but not with cholesterol or progesterone. This procedure reverses the standard strategy for synthesizing ester linkages in hapten-protein conjugates and should have widespread applicability for the preparation of other such conjugates for immunological work.
D000818:Animals; D002417:Cattle; D004440:Ecdysone / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000494:pharmacology; D006241:Haptens / Q000138:chemical synthesis*; D007106:Immune Sera; D011485:Protein Binding; D011817:Rabbits / Q000276:immunology; D011863:Radioimmunoassay; D013387:Succinic Anhydrides / Q000494:pharmacology; D013954:Thyroglobulin / Q000378:metabolism*
D006241:Haptens; D007106:Immune Sera; D013387:Succinic Anhydrides; D004440:Ecdysone; D013954:Thyroglobulin; D005022:Ethyldimethylaminopropyl Carbodiimide