CAJ | 학술논문

目的：建立酸性橙Ⅱ半抗原及人工抗原的合成及鉴定方法。方法将重氮化后的2-氨基-5-磺基苯甲酸与2-萘酚经偶氮反应合成带羧基基团酸性橙Ⅱ,通过碳二亚胺法将其与载体蛋白(BSA)偶联制备酸性橙Ⅱ人工抗原。采用核磁共振碳谱和氢谱对合成的酸性橙Ⅱ半抗原进行结构分析,并用紫外光谱扫描法和蛋白质电泳法鉴定酸性橙Ⅱ人工抗原并计算偶联比。结果通过偶氮反应及碳二亚胺法最终得到纯化后的酸性橙Ⅱ半抗原约47 mg,核磁结果显示成功合成酸性橙Ⅱ半抗原。经SDS-PAGE、N-PAGE和紫外光谱扫描结果均显示酸性橙Ⅱ人工抗原蛋白分子质量大于 BSA,所带负电荷大于 BSA,且合成后的酸性橙Ⅱ人工抗原具有吸收峰叠加效应,均表明酸性橙Ⅱ人工抗原合成成功,偶联比为20.9：1。结论此方法能够成功合成酸性橙Ⅱ半抗原和人工抗原,为进一步制备特异性酸性橙Ⅱ抗体和建立酸性橙Ⅱ的免疫检测方法奠定基础。
목적：건립산성등Ⅱ반항원급인공항원적합성급감정방법。방법장중담화후적2-안기-5-광기분갑산여2-내분경우담반응합성대최기기단산성등Ⅱ,통과탄이아알법장기여재체단백(BSA)우련제비산성등Ⅱ인공항원。채용핵자공진탄보화경보대합성적산성등Ⅱ반항원진행결구분석,병용자외광보소묘법화단백질전영법감정산성등Ⅱ인공항원병계산우련비。결과통과우담반응급탄이아알법최종득도순화후적산성등Ⅱ반항원약47 mg,핵자결과현시성공합성산성등Ⅱ반항원。경SDS-PAGE、N-PAGE화자외광보소묘결과균현시산성등Ⅱ인공항원단백분자질량대우 BSA,소대부전하대우 BSA,차합성후적산성등Ⅱ인공항원구유흡수봉첩가효응,균표명산성등Ⅱ인공항원합성성공,우련비위20.9：1。결론차방법능구성공합성산성등Ⅱ반항원화인공항원,위진일보제비특이성산성등Ⅱ항체화건립산성등Ⅱ적면역검측방법전정기출。
Objective To explore a method for the synthesis and identification of acid orange Ⅱhapten and artificial antigen. Methods Acid orange Ⅱ hapten was synthesized by the reaction between 2-amino-5-sulfo-benzoic acid and 2-naphthol, then acid orange Ⅱ artificial antigen was prepared by active ester method. Acid orange Ⅱ hapten was identified by mass spectrometry and acid orange Ⅱ artificial antigen was identified by UV spectrophotometric, SDS-PAGE as well as N-PAGE. Results The structure of acid orange Ⅱ hapten was consistent with the predictive structure by mass spectral analysis. By the methods of SDS-PAGE and N-PAGE, the molecule weight and negatively charged electrons of acid orange Ⅱ artificial antigen were greater than BSA. Acid orange Ⅱ artificial antigen showed the superposition phenomenon by Ultraviolet spectrum. According to the UV, acid orange Ⅱ hapten and BSA coupling ratio was 20.9:1. Conclusion This method can be applied for acid orange Ⅱ hapten and artificial antigen synthesis and provide technical basis for the preparation of acid orange Ⅱ specificity antibodies and development of acid orange Ⅱ immune detection.