氯霉素全抗原的合成与鉴定研究.pdfVIP

M037 氯霉素全抗原的合成与鉴定 吴媛媛 包永明 王龑 梁兴超 修志龙 (大连理工大学环境与生命学院生物科学与工程系，大连 116024) 摘 要 采用戊二醛法和羰基二咪唑法，将半抗原氯霉素(CAP)与载体牛血清白蛋白(BSA)偶联， 制得氯霉素全抗原 CAP-BSA，其紫外扫描光谱和红外光谱表明偶联成功。所合成的抗原采用聚 丙烯酰胺凝胶电泳(SDS)只见一条区带，表明所合成的氯霉素与 BSA 偶联物可以做为免疫抗原。 用所买的氯霉素快速检测试剂盒可以定量测定所合成抗原中氯霉素的含量。将固体配成 1mg/ml 溶液，用 TNBS 法测氨基酸残基，得到 BSA 上偶联的氯霉素数目。 关键词 氯霉素 牛血清白蛋白 戊二醛 羰基二咪唑 SYNTHESIS AND IDENTIFICATION OF IMMUNOGEN CHORAMPHENICOL WU Yuanyuan, BAO Yongming, WANG Yan, LIANG Xingchao and XIU Zhilong (Department of Bioscience and Biotechnology, School of Environmental and Biological Science and Technology, Dalian University of Technology, Dalian,116024 ) Abstract In order to establish an enzyme-linked immunosorbent assay (ELISA) for rapidly examing residual chloramphenicol (CAP) in food, the antibody with high titer and specificity must be firstly prepared. So it was necessary for antibody production to conjugate the hapten (CAP) with a carrier protein to synthesize immunogen (antigen). Chloramphenicol was coupled with carrier protein bovine serum albumin (BSA) to prepare complete antigen CAP-BSA by glutaraldehyde (GA) and N-N’-carbonyldiimidazole (CDI) in this paper. The successful linkage of the CAP-BSA was identified by the ultraviolet spectrum (UV) and the infrared spectrometry (IRS). There was only one zone in SDSand the products could be considered as the immunogen. An ELISA kit had been used to determine the content of CAP in the CAP-BSA. The residual amount of amino groups was determined by the TNBS spectrometry, and the amount of CAP linked to BSA was achieved. All the above results indicated that the immunogen CAP-BSA was