Production of highly specific polyclonal and monoclonal antibodies using estradiol-3-O-carboxymethyl ether as hapten.
The preparation of high affinity and high specificity polyclonal and monoclonal antibodies to estradiol is described. Monoclonal antibodies were derived from BALB/c mice hyperimmunized with estradiol-3-O-carboxymethyl ether conjugated to bovine serum albumin. Spleen cells were hybridized with mouse myeloma cells. Quite a few monoclonal antibodies showed very good affinity for estradiol. Extended immunization and hyperimmunization were essential for producing a greater number of positive clones secreting high affinity antibodies. Binding constants of the antisera and their cross-reactivities with related steroids were calculated. Both polyclonal and monoclonal antibodies showed very high affinity for estradiol exhibiting little or no cross-reactivities with structurally related steroids indicating that this site of linkage is a good choice for discriminating between differences at the 16-17 position in the D-ring. This monoclonal antibody (44.28.6), having negligible cross-reactivity with estriol and estrone, can be used for diagnostic purposes.
Mandal
C
C
Indian Institute of Chemical Biology, Calcutta.
Ali
N
N
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000915:Antibody Affinity; D000918:Antibody Specificity; D002459:Cell Fusion; D003429:Cross Reactions; D004958:Estradiol / Q000031:analogs & derivatives* / Q000276:immunology; D005260:Female; D006241:Haptens / Q000276:immunology*; D007106:Immune Sera / Q000276:immunology*; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000276:immunology; D008738:Methyl Ethers / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D012710:Serum Albumin, Bovine / Q000276:immunology; D013154:Spleen / Q000166:cytology; D013256:Steroids / Q000276:immunology
D000911:Antibodies, Monoclonal; D006241:Haptens; D007106:Immune Sera; D007074:Immunoglobulin G; D008738:Methyl Ethers; D013256:Steroids; D012710:Serum Albumin, Bovine; C058989:estradiol-3-O-carboxymethyl ether; D004958:Estradiol