An enzyme-linked immunoassay for the detection of medroxyprogesterone acetate in intestines based on monoclonal antibody |
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| Authors: | Hongli Qi Ruirui Wang Guang Yang Jinlan Liu |
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| Affiliation: | 1. Ocean University of China , Qingdao, 266003, P.R. China;2. Key Laboratory of Aquatic Ecology and Aquaculture of Tianjin , Tianjin Agricultural University , Jinjing Road 22, Xiqing District, Tianjin, 300384, P.R. China;3. National Marine Environmental Monitoring Center , Dalian, 116023, P.R. China;4. Key Laboratory of Aquatic Ecology and Aquaculture of Tianjin , Tianjin Agricultural University , Jinjing Road 22, Xiqing District, Tianjin, 300384, P.R. China |
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| Abstract: | A competitive indirect enzyme-linked immunoassay (ciELISA) based on a monoclonal antibody was developed to detect medroxyprogesterone acetate (MPA) in sheep intestines, which are used as sausage casings. MPA has a relative molecular mass of only 344.5, and it has no immunogenicity. By activating MPA with o-carboxymethyl hydroxylamine hemi HCl, a hapten of MPA (3-CMO-MPA) was synthesised and linked to bovine serum albumin (BSA) and ovalbumin (OVA) to prepare artificial antigens. The monoclonal antibody was prepared from mice immunised with MPA-BSA. The ELISA method allowed MPA detection in a range of 0.1–24.3 ng/ml with an average IC50 value of 1.7 ng/ml. The recoveries of MPA from spiked intestinal tissues at levels of 0.5–5 ng/g ranged from 85.2% to 115.1% with CVs of 8.8–11.2%, and the detection limits were 0.5 ng/g in intestinal tissues. The ciELISA test proved to be a rapid, sensitive, economical and reliable method for screening MPA residues in intestinal tissues. |
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| Keywords: | ciELISA medroxyprogesterone acetate monoclonal antibody detection |
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