A simple procedure for enzyme labelling of progesterone derivatives: application of active esters formed using N,N'-disuccinimidyl carbonate.

ABSTRACT

N,N'-Disuccinimidyl carbonate was used to synthesize N-hydroxysuccinimide esters of 11 alpha-hydroxyprogesterone 11-[1,4-14C]hemisuccinate (P11-HS) and 11 alpha-hydroxyprogesterone 11-glucuronide (P11-Glu) in a one-step procedure at room temperature. Enzyme-labelled progesterone was subsequently formed by reaction of the ester, without purification, with alkaline phosphatase. Labels produced by this simple procedure compared favourably with those formed using an established method of ester synthesis when assessed in enzyme immunoassay (EIA).