羟基红花黄色素A人工抗原的制备

Preparation of artificial antigen of hydroxysafflor yellow A

目的研究羟基红花黄色素A(HSYA)与牛血清白蛋白(BSA)和卵清蛋白(OVA)的偶联方法,获得具有免疫原性的羟基红花黄色素A人工抗原。方法通过偶联物透析液的颜色变化、紫外图谱、红外扫描图谱及动物免疫试验来鉴定制备的人工抗原。结果HSYA-BSA人工抗原在紫外扫描中表现出HSYA和BSA的特征吸收峰;在红外扫描中HSYA-BSA具有HSYA特征官能团的吸收峰和氨基酸的特征吸收峰;间接ELISA检测结果显示产生抗HSYA的特异性抗体,血清效价可达1∶3 200;标准曲线方程计算得BSA和HSYA的结合率是1∶50。结论通过直接偶联法成功制备出具有免疫原性的羟基红花黄色素A人工抗原。

Objective To develop the proper method for conjugating hydroxysafflor yellow A （HSYA） with carrier protein, bovine serum albumin （BSA） and ovalbumin （OVA）, as well as preparation of artificial immunogen of HSYA. Methods Immunogen HSYA-BSA and coating antigen HSYA-OVA were prepared by immediate coupling method, and the characterizations of the artificial antigen were studied by color change, ultraviolet, and infrared radiation spectra shift, and mice immunization of the conjugate. Results HSYA-BSA Artificial antigen exhibited characteristic ultraviolet spectra of BSA and HSYA and showed obvious absorption peak in characteristic functional group of HSYA and amino acid in infrared radiation spectra. The immunized mice antiserum specifically bound the HSYA; The specific antibody for HSYA was determined by indirect ELISA. The determination of antiserum was 1 ： 3 200. The conjugating rate between BSA and HSYA was 1 ： 50 calculated by regression equation. Conclusion The artificial immunogen of HSYA is synthesized by immediate coupling method successfully.