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Phase Separation Immunoassay of Methyltestosterone

Gao Jun; Li Yuanzong; Chang Wenbao; Ci Yunxiang (Department of Chemistry,Peking University,Beijing 100871 )  
A notel immunoassay of methyltestosterone (MT) in serum was developed. The antigen of MT (AgMT) was synthesized by covalently bonding MT to bovine serum albumin (BSA). The polycolnal antibody of AgMT (AbMT ) was raised from rabit. AgMT was conjugated to poly-N-isopropylacrylamide (PNIP) to form AgMT bound thermally reversible hydrogel PNIP-AgMT. A competitive immunoassay method based on the competition of PNIP-AgMT and free MT with limited amount of horseradish peroxidase(HRP) labeled AbMT was established. The separation of PNIP in the assay process was achieved by precipitation of the immuno-complex above its critical solution temperature. The amount of MT was determined by measuring the initial rate of the oxidation of 4-aminoantipyrine by H2O2 catalyzed by HRP-immuno- complex precipitated. The linear range for MT was 50 μg/L ~ 7. 5 mg/L and the detection limit was 50 μg/L. The recoveries of MT form human serum were satisfactory.
【Fund】: 国家自然科学基金!29775002
【CateGory Index】: O629
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