A sensitive bridge heterologous enzyme immunoassay of progesterone using geometrical isomers

Abstract

A sensitive bridge heterologous enzyme immunoassay of progesterone using geometrical isomers of progesterone 3( O-carboxymethyl)oxime( E/ Z) was developed. Isomers were separated by synthesis of N-hydroxysuccinimide esters. Progesterone 3(Z)( O-carboxymethyl)oxime N-hydroxy-succinimide ester bound with β-galactosidase in an appropriate molar ratio provided a conjugate suitable for an enzyme immunoassay. The antiserum was raised in rabbits by immunizing the animals with the progesterone 3( E)( O-carboxymethyl)oxime-bovine serum albumin conjugate. This bridge heterologous enzyme immunoassay proved to have sufficient sensitivity equivalent to radioimmunoassay and excellent specificity.