异氰酸酯类代表物人工抗原的合成及鉴定

Synthesis and identification of artificial antigen for the isocyanates

目的合成异氰酸酯类物质二异氰酸甲苯酯(2,4-TDI),二苯基甲烷二异氰酸酯(MDI)及环己烷二异氰酸酯(HDI)的人工完全抗原,为检测环境中异氰酸酯类物质的免疫分析方法的建立奠定基础。方法将不同浓度TDI,MDI,HDI连接到载体蛋白HSA(人血清白蛋白)和OVA(卵清蛋白)上,采用紫外全谱扫描,三硝基苯磺酸法(TNBS)和SDS-PAGE(聚丙烯酰胺凝胶电泳)对合成抗原进行鉴定。结果合成的TDI、HDI、MDI完全抗原的紫外吸收光谱、SDS-PAGE电泳条带呈现出与半抗原、载体蛋白不同的吸收峰和条带特性,说明半抗原与载体蛋白偶联成功,TNBS法鉴定结果显示成功合成具有不同偶联比的TDI、HDI、MDI的各3种免疫原和包被原。结论本研究成功合成TDI、MDI、HDI的人工完全抗原,为其免疫测定方法的建立提供了条件。

Objective To synthesize the artificial complete antigen of isocyanates including toluene diisocyanate （TDI）, methyl- ene diphenyl diisocyanate（HDI） and cyclohexyl diisocyanate（MDI）. It is conducive to establish the immunoassay techniques of isocyanates in environment. Methods TDI, HDI and MDI at different concentrations were conjugated into human serum albu- ,nin（HSA） and ovalbumin （OVA） to prepare artificial complete antigen. Then uhraviolet spectrum scan, TNBS and SDS- PAGE were used to identify these synthesized antigen. Results The uhraviolet absorption spectrum and SDS -PAGE bands of artificial complete antigens showed different absorption peaks and band characteristics with those of TDI, HDI, MD! as well as carrier proteins, indicating the successful coupling of hapten and carrier protein. The identification results of TNBS method showed that artificial antigens with different coupling ratios were successfully acquired. Conclusion This study has successfully synthesized artificial complete antigens of TDI, HDI and MDI. It was helpful to establish a sensitive and active immunoassay technique.