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Development of Monoclonal Antibody‐based Enzyme‐linked Immunosorbent Assay to the Estrogen Diethylstilbestrol
Author(s) -
Wang WenJun,
Li Ji,
Zhao JiXun,
Zhang GuoZhong,
Xu Ting,
Sheng Wei
Publication year - 2006
Publication title -
chinese journal of chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.28
H-Index - 41
eISSN - 1614-7065
pISSN - 1001-604X
DOI - 10.1002/cjoc.200690329
Subject(s) - chemistry , diethylstilbestrol , immunogen , hapten , hexestrol , polyclonal antibodies , immunoassay , chromatography , monoclonal antibody , detection limit , bovine serum albumin , estrogen , enzyme , metabolite , antibody , biochemistry , endocrinology , hormone , immunology , medicine , biology
Diethylstilbestrol (DES) is a synthetic estrogen that has ever been used worldwide. Polyclonal antibodies (PAbs) were used in immunoassay for detection of DES residues in environmental and agricultural samples in previous paper. In this paper, an indirect competitive enzyme‐linked immunosorbent assay (icELISA) was developed based on monoclonal antibody (MAb) for the determination of diethylstilbestrol. Mono‐ o ‐carboxypropyldiethylstilbestrol (DES‐CP) and mono‐ o ‐carboxymethyldiethylstilbestrol (DES‐CME) were synthesized to be haptens. DES‐CP was coupled to bovine serum albumin (BSA) to be an immunogen in BALB/c female mouse for MAb production. The MAb was characterized for specificity and affinity to DES in icELISA. Under the optimum condition, the icELISA showed an IC 50 of 9.8 ng/mL, the limit of detection (IC 20 ) of 2.3 ng/mL and a working range of 2–42 ng/mL. Hexestrol and dienestrol exhibited cross‐reactivity values were 44% and 27%, respectively. Cross‐reactivity of natural estrogen 17 β ‐estradiol was less than 0.1%. The influences of some factors such as salt concentration, pH and organic solvent concentration on the assay were evaluated. The concentrations of DES in the fortified water samples determined by the assay were correlated well with the fortification levels. The results were confirmed with analysis by HPLC.