玉米赤霉烯酮半抗原及全抗原的合成与鉴定

Synthesis and identification of the hapten and complete antigens for Zearalenone

目的:合成玉米赤霉烯酮(ZEN)半抗原及全抗原,并鉴定其是否合成成功,为制备ZEN抗体奠定基础。方法:将ZEN与羧甲氧基胺半盐酸盐(O-(carboxym ethyl)hydroxyla-m ine hem ihydrochloride)反应,合成半抗原ZEN-6-羧甲氧基胺(ZEN-oxim e),采用TLC、HPLC和液相-质谱联用技术进行半抗原的纯化、鉴定;通过活泼酯法将半抗原与载体蛋白偶联制备ZEN全抗原,采用紫外光谱法、TNBS法结合比的测定以及免疫学方法进行全抗原鉴定。结果:鉴定结果表明目标半抗原及全抗原合成成功。结论:本研究采用多种方法进行半抗原及全抗原的鉴定,结果皆表明目标半抗原及全抗原合成成功,说明本研究合成ZEN半抗原及人工抗原的技术路线是可行的,为进一步研制ZEN的抗体奠定良好基础。

AIM： To synthesize and identify the hapten and complete antigens for Zearalenone for the purpose of preparation of anti-Zearalenone antibodies.METHODS： Zearalenone and carboxymethoxylamine hemihydrochloride were incubated to synthesize the hapten ZEN-oxime.The ZEN-oxime was identified by thin layer chromatography（TLC）,high performance liquid chromatographic（HPLC） and liquid chromatography-tandem mass spectrometry（LC-MSn）.The ZEN-oxime was further conjugated to bovine serum albumin（BSA） or ovalbumin（OVA） to form the complete antigens by NHS ester method.The complete antigens were identified by ultraviolet spectrum,2,4,6-trinitrobenzene sulfonicacid method（TNBS） and immunological analysis.RESULTS： The results indicated that the hapten and complete antigens were prepared successfully.CONCLUSION： The hapten and complete antigens for Zearalenone have been successfully synthesized and identified by several methods,which enables further preparation of anti-Zearalenone antibodies.