羟基红花黄色素A人工抗原两种制备方法的比较  

Comparation of the preparation of Hydroxysafflor yellow A artificial antigen between immediate and sodium iodide coupling method

作　　者：侯世斌[1] 吕芳[1] 苏幼红[1] 李江伟[1] 张富春[1] 

机构地区：[1]新疆大学生命科学与技术学院新疆生物资源基因工程重点实验室,乌鲁木齐830046

出　　处：《食品科技》2008年第10期218-222,共5页Food Science and Technology

基　　金：科技部省部共建新疆生物资源基因工程重点实验室开放基金(XJDX0201-2006-04)

摘　　要：目的:对过碘酸钠法和直接法制备的羟基红花黄色素A全抗原进行比较,以期获取理想的羟A人工抗原。方法:通过透析液的颜色变化、紫外扫描、红外扫描及动物免疫试验等比较两种方法合成的人工抗原特点。结果:两种方法制备的人工抗原在紫外扫描中都有蛋白的特征吸收峰,只是受偶联基团的影响特征峰都蓝移到275nm附近,但是直接法的400nm处羟A特征峰保留,过碘酸钠法的消失。对直接法的偶联物进行红外扫描,显示羟A特征官能团的吸收峰和蛋白的特征吸收峰。过碘酸钠法的经过五免老鼠仍然监测不到针对羟A的特异性抗体,用直接法的免疫抗原继续免疫二次产生了抗羟A的特异性抗体,三免血清效价达1:3200;用直接法的免疫两只新西兰大白兔六免抗HSYA抗体效价达1:3200以上。结论:直接法适合制备羟A人工抗原,可以为后续研究奠定可靠的前提条件。Objective： To develop efficient method for conjugating HSYA with carrier protein BSA, and preparation of artificial immunogen of hydroxysafflor yellow A （HSYA）, we compared two preparing methods of sodium iodide and direct conjugating. Method： Immunogen HSYA-BSA and coating antigen HSYA-OVA were prepared by the two methods, and the characterizations of artificial antigen were studied by color changes, ultraviolet and infrared radiation spectra shift and mice （rabbits）immunization of the conjugate. Results： The Artificial antigen prepared by the two methods both exhibited characteristic ultraviolet spectra of BSA, but the characteristic peaks both moved to near 275 nm by the impact of coupling groups, and there is a characteristic absorption peak of HSYA in immediate coupling, but never in the sodium iodide method. HSYA-BSA in immediate coupling showed obvious absorption peak of characteristic functional group of HSYA and amino acid in infrared radiation spectra. No antibody specifically bound the HSYA was detected in the sera of mouse immunizated with artificial antigens synthesized by the sodium iodide. In contrast, antibody against HSYA conjugated with BSA by immediate coupling was detected, and the titer of determined antiserum was 1：3200.The similar result was obtained in immunized New Zealand rabbits with antigens prepared of immediate coupling method, and the titer of determined antiserum was over 1：3200 after the sixth immunization. Conclusion： The artificial immunogen of hydroxyl safflor yellow A synthesized by immediate coupling method was more efficient than the sodium iodide method and reliably for the follow-up research.

关 键 词：羟基红花黄色素A 人工抗原 过碘酸钠法 直接法 间接ELISA 

分 类 号：R927.2[医药卫生—药学] TS202.3[轻工技术与工程—食品科学]