Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Estradiol

doi:10.7506/spkx1002-6630-201208030

Abstract

Abstract: In order to establish a simple and fast assay for the detection of estradiol in foods, estradiol hapten was synthesized and identified by nuclear magnetic resonance (NMR) and infrared spectroscopy. The hapten was activated by DCC/NHS ester method and covalently coupled to carrier proteins (bovine serum albumin and ovalbumin). Polyclonal antibodies were obtained from immunized rabbits. The optimal reaction conditions of indirect competitive enzyme-linked immunosorbent assay (icELISA) were established. The IC50 and LOD of the developed method were 3.9 ng/mL and 0.3 ng/mL, respectively. The cross-reactivity with one of 8 structural analogues of estradiol was less than 1.21%. High sensitivity and specificity will contribute to the development of commercial ELISA kits for the detection of estradiol in foods.

Key words: estradiol, antibody, indirect competitive enzyme-linked immunosorbent assay

CLC Number:

TS201.3

LIUJin，LEIHong-tao，SUNYuan-ming，LIZhen-feng，YANGJin-yi，SHENYu-dong，WANGHong，ZENGDao-ping. Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Estradiol[J]. FOOD SCIENCE, 2012, 33(8): 146-150.

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Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Estradiol

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