Homogeneous enzyme immunoassay of estradiol using estradiol-3-O-carboxymethyl ether as hapten.
A homogeneous enzyme immunoassay for estradiol estimation has been developed, which can be extended to other steroids. A new procedure for the preparation of estradiol -3-0- carboxymethyl ether by a simple one step reaction in high yield (90%) has been described. This hapten has been used for raising highly specific anti-estradiol antibody in rabbits and for preparation of enzyme conjugates. Two different enzymes, lysozyme and glucose -6- phosphate dehydrogenase have been studied for their suitability as enzyme labels. Our results indicate that lysozyme-conjugate meets the essential requirement for a practical enzyme immunoassay. The advantage of the present nonradioactive procedure is the overall simplicity, low cost and high stability of the reagents.
Dhar
T K
TK
Indian Institute of Chemical Biology, Calcutta.
Samanta
A K
AK
Ali
E
E
D002021:Buffers; D004958:Estradiol / Q000031:analogs & derivatives* / Q000032:analysis* / Q000138:chemical synthesis; D005954:Glucosephosphate Dehydrogenase; D006241:Haptens; D007124:Immunoenzyme Techniques; D008738:Methyl Ethers / Q000138:chemical synthesis; D009113:Muramidase
D002021:Buffers; D006241:Haptens; D008738:Methyl Ethers; C058989:estradiol-3-O-carboxymethyl ether; D004958:Estradiol; D005954:Glucosephosphate Dehydrogenase; D009113:Muramidase