磺胺类药物单链抗体的制备及其与磺胺噻唑相互作用研究  

Construction of a Single Chain Variable Fragment Antibody Against Sulfonamides and Its Interaction with Sulfathiazole

作　　者：李永翰 温凯[1] 于雪芝[1] 李成龙[1] 王战辉[1,2] 

机构地区：[1]中国农业大学动物医学院,北京100193 [2]动物源食品安全检测技术北京市重点实验室,北京100193

出　　处：《分析化学》2015年第3期366-370,共5页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金面上项目(No.31372475)资助~~

摘　　要：抗体与对应的小分子待测物之间的相互作用模式决定了免疫分析的特性。本研究以磺胺类药物杂交瘤细胞株4C7为起点,应用基因工程技术制备出单链抗体sc Fv4C7,采用间接竞争ELISA法对比其与母本单克隆抗体的识别特性,同时采用同源建模构建sc Fv4C7的三维立体结构,并与磺胺噻唑(STZ)进行分子对接。间接竞争ELISA结果显示sc Fv4C7保留了亲本单克隆抗体的识别特性,分子对接结果显示STZ深陷入抗体的重链和轻链形成的"口袋"中,STZ分子更靠近重链,且主要与抗原互补决定区CDR H3相互作用。本研究为制备识别谱更广、亲和力更高的磺胺类药物抗体提供了必要的结构信息。The interaction between the antibody and the corresponding target molecule determines the characteristics of immunoassay. In this study,a single chain variable fragment antibody（ sc Fv4C7） derived from the hybridoma strain 4C7 were prepared via genetic engineering technique. The recognition properties of sc Fv4C7 was determined and compared to those of the parent monoclonal antibody by indirect competitive enzyme-linked immunosorbent assay（ ic-ELISA）. Three dimensional structure of the sc Fv4C7 was presented by Swiss-Model,and sulfathiazole（ STZ） was docked to the sc Fv4C7 model to obtain the structure of the binding complex. The results from the ic-ELISA showed that the binding properties of sc Fv4C7 were comparable with the parent monoclonal antibody and STZ was almost completely buried in a deep binding pocket formed by the heavy chain and light chain of the antibody. The interaction between STZ and sc Fv4C7 was more closely related to the heavy chain and the complementarity-determining region（ CDR） H3 loop played more important role than other CDR loops. The study preliminary provided the necessary structural information for the preparation of antibody with broader specificity and higher affinity.

关 键 词：相互作用 同源建模 分子对接 单链抗体 磺胺噻唑 

分 类 号：TQ460.1[化学工程—制药化工]