雌二醇单克隆抗体制备及间接竞争ELISA检测方法的建立

doi:10.16431/j.cnki.1671-7236.2017.10.038

Abstract

Abstract:

In order to establish a sensitive,specific and rapid method for the detection of estradiol (E₂) residues, an indirect competitive enzyme-linked immunosorbent assay (icELISA) based on monoclonal antibody (MAb) for E₂ was developed. BALB/c mice were immunized by E₂-BSA and cell fusion technology was employed to screen hybridoma cell lines. One hybridoma cell line (3H3) was isolated, which produced monoclonal antibody that could binding E₂. Under the optimized conditions, the icELISA based on 3H3 for E₂ showed a half maximum inhibition concentration (IC₅₀) values of 1.636 ng/mL and detection ranges of 0.202 to 13.281 ng/mL with cross-reactivities for estriol and ethinyloestradiol of 0.31% and 0.25%, respectively, and negligible cross-reactivities with other E₂ analogs including estrone, estradiol valerate, estradiol benzoate, quinestrol, diethylstilbestrol and nonylphenol. The results demonstrated that the developed method could meet the requirements of high sensitivity detection of E₂ residue in food samples.

Key words: estradiol; monoclonal antibody; indirect competitive ELISA

CLC Number:

S859.84

BAI Yu, HU Jing-yan, XU Yong-peng, HE Jie, JIN Jun-jie, LI Pei-de, LIU Su-zhen. Preparation of Monoclonal Antibody and Establishment of Indirect Competitive ELISA of Estradiol[J]. , 2017, 44(10): 3100-3105.

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Preparation of Monoclonal Antibody and Establishment of Indirect Competitive ELISA of Estradiol

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