兼具酶催化活性的布洛芬人工抗原的制备与应用初探

Preparation and primary application of ibuprofen artificial antigen with catalytic activity

目的:制备以辣根过氧化物酶为载体蛋白的布洛芬人工抗原,并快速测定其偶联比、酶催化活性,进而应用于尿液中布洛芬的检测。方法:利用活化酯法将布洛芬与载体蛋白辣根过氧化物酶偶联,制备布洛芬的人工抗原,采用生物质谱法完成人工抗原偶联比的测定,并对酶催化活性进行了验证。结果:布洛芬人工抗原偶联比为5,辣根过氧化物酶的催化活性未受到任何影响,可以应用于尿液中布洛芬的检测。结论:以酶作为载体蛋白的人工抗原兼具免疫原和酶催化的双重性质,为免疫分析提供了一种新的简单可行方法。

OBJECTIVE To prepare ibuprofen artificial antigen and fast determine the coupling ratio of ibuprofen and biological macromolecules.METHODS The ibuprofen artificial antigen was prepared by using active ester method to couple ibuprofen to horseradish peroxidsae（HRP）,and its coupling ratio was determined by using biological mass spectrometry method.Also,the original catalytic activity of the artificial antigen was verified.RESULTS The coupling ratio of ibuprofen artificial antigen was five and the catalytic activity of the artificial antigen was the same as that of HRP.CONCLUSION The operation is simple by using the biological mass spectrometric method to determine the coupling ratio of artificial antigen.The coupling ratio of ibuprofen and HRP can get only within a few seconds.Furthermore,the antigen has catalytic activity so it can provide a simple and feasible method for immunoassay.