罂粟碱和阿特拉津的蛋白芯片研究与实验条件优化

目的探索和优化罂粟碱抗原芯片和阿特拉津抗体芯片的制备及相关实验条件,利用抗原抗体特异性结合作用达到对罂粟碱和阿特拉津分子定性、定量检测的目的。方法将罂粟碱和农药阿特拉津与蛋白偶联后的衍生物作为完全抗原,与其相应的特异性抗体结合,从而间接实现对小分子物质的检测。结果当以醛基化玻片为固体载体时,罂粟碱和阿特拉津分别以2%卵清蛋白(OVA)和3%牛血清蛋白(BSA)最适合作封闭剂;当pH 7.2~7.4之间、抗原抗体反应时间120 min、反应温度为37℃和适度漂洗时能获得理想荧光信号。结论罂粟碱和阿特拉津蛋白衍生物完全能与其相应的抗体进行特异性结合,荧光信号随待测物浓度的降低而增强。

Optimization of protein chip technologies for the detection of atrazine and paraverine

Objective To study the preparation of a chip of paraverine antigen or anti-atrazine antibody that recognizes its partner qualitatively and quantitatively with high affinity and specificity under optimized conditions.Methods The derivatives of pesticide atrazine and paraverine were successfully synthesized in this study,and then conjugated to ovalbumin,used as antigen which proved subsequently high specificity against its corresponding antibody.Results When aldehydized glass slide was used as a carrier,atrazine and paraverine were well blocked with 2% ovalbumin and 3% BSA respectively.Atrazine's fluorescence intensity was best when the antibody concentration was reached to 0.254 mg/ml,and 3% BSA(pH 7.2~9.0) was used as buffer blocking for 30~60 minutes.Conclusions The protein derivatives of atrazine and paraverine are able to conjugate specifically with their antibodies.The fluorescence intensity increases along with decreasing concentration of analytes.