氯霉素全抗原的合成与鉴定_吴媛媛.docVIP

· 化学通报 　2005 年 第 6 期 　　　　　　　　　　http :ΠΠwww . hxtb. org 458 · 氯霉素全抗原的合成与鉴定 3 吴媛媛 　包永明 　王　 　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 CA P2B SA ,并由紫外光谱和红外光谱确定偶联成功。所合成抗原的聚丙烯酰胺凝胶 电泳( SDS) 只见一条区带 ,表明所合成的氯霉素与 BSA 偶联物可以作为免疫抗原。用市售的氯霉素快速 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1mgΠmL溶液 ,用 TNB S 法测氨基酸残 基 ,得到 B SA 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 Identif ica tion of Chora mphen icol Immunogen 3 Wu Yua nyua n ,Bao Yongmi ng ,Wang Yan ,Liang Xi ngc hao ,Xiu Zhilong (Department of Bioscience and Biotechnology , School of Environmental and Biological Science and Technology , Dalian University of Technology , Dalian116024) Abstract 　In order to establish an enzyme2linked immunosorbent assay ( EL ISA) for rapidly examing residual chloramphenicol ( CAP) in food , the antibody with high titer and specificity must be firstly prepared. So it was necessaryfor 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 CA P2B SA by glutaraldehyde ( GA) and N ,N′2carbonyldiimidazole ( CDI) . The successful linkage of the CA P2 BSA was identified by UV and IR. There was only one zone in SD S2PA GE and the products could be considered as the immunogen. An EL ISA kit had been used to determine the content of CAP in the CA P2BSA. The residual amount of amino group s was determined by the TNB S spectrometry , and the amount of CAP linked to BSA was achieved. All the above results indicated that the immunogen CA P2B SA was successfully synthesized , which was goi ng to be injected in the animals to obtain CAP antibody. Ke y wor ds 　Chloramphenicol ,Bovine serum albumin , Glutaraldehyde , N ,N′2Carbonyldiimidazole 氯霉素(Chloramphenicol ,CAP) 是应用广泛的抗生素 ,对畜禽疾病的控制和治疗发挥了重要作 用。但由于氯霉素存在严重的副作用 ,能引起人的再生障碍性贫血、粒状白细胞缺乏症等疾病 [1 ] , 其在动物的肉、奶、蛋中的残留 ,威胁着人类的健康。欧美等发达国家已相继禁止或严禁使用 ,并对 畜产品中