3-硝基酪氨酸人工抗原的合成与鉴定  

Synthesis and Identification of the Antigens for the 3-nitrotyrosine

作　　者：彭方毅[1,2] 杨海艳[1] 姜海蓉[1] 徐建建[1] 伍娟 周欢[1] 袁兵占[1] 

机构地区：[1]重庆理工大学药学与生物工程学院,重庆400054 [2]重庆市江津区中心医院,重庆402260

出　　处：《重庆理工大学学报（自然科学）》2013年第2期28-31,44,共5页Journal of Chongqing University of Technology：Natural Science

基　　金：重庆市自然科学基金资助项目(2008BB5331;2009BB7231);教育部重点资助项目(210178)

摘　　要：通过戊二醛法将3-硝基酪氨酸与卵白蛋白(OVA)偶联,制备人工抗原,并用作免疫抗原。用紫外扫描法和聚丙烯酰胺电泳法对其进行鉴定,用BCA法测定二者结合物的质量体积分数,并测得结合物的偶联率。3NT与OVA结合物的质量体积分数为1.69 mg/mL。SDS-PAGE电泳测定其分子量大于43KD。紫外分光光度法测定二者结合物与游离的3NT与游离的OVA之间存在漂移,说明3NT偶联到结合物上,估算其偶联率为5.4。用结合物免疫BALB/C小鼠,间接ELISA测定多抗上清(pAb)效价,获得了高效价的pAb。In this study, Ovalbumin （OVA） was used as a protein carrier to couple with 3-nitroty- rosine by glutaraldehyde method. The artificial antigen for 3-nitrotyrosine was thus prepared as immu- noantigen in enzyme-linked immunosorbent assay （ELISA） protocol; Ultraviolet spectroph- otometry and sodium dodecyl-sulfatepolycrylamide gel electrophonesis （SDS-PAGE） were applied to determine it, and the concentration of the conjugate was determined by BCA method and the conjugated ratio was estimated. The concentration of the conjugate was 1.69 mg/mL; the molecular weight was more than 43KD by SDS-PAGE. Based on the UV spectrum, the excursion of the conjugate was different from 3NT and OVA, that is to say, the conjugate was succeed and the coupling rate was about 5.4,the high titer anti-3NT pAb was been produced in serum of immunized BALB/C mice. The results showed that the artificial antigen for 3-nitrotyrosine was synthesized successfully, which made it possi- ble to establish method of 3NT residues in diseases

关 键 词：3-硝基酪氨酸 卵白蛋白 戊二醛法 

分 类 号：R392.33[医药卫生—免疫学]