抗乳铁蛋白单链抗体制备及鉴定  

Preparation and Identification of Anti-lactoferrin Single Chain Fv Antibody

作　　者：虞东芳[1] 刘祖国[1] 李民友[1,2] 顾熊飞[1,2] 胡皎月[1] 

机构地区：[1]广东省中山大学眼表疾病研究中心,中山大学中山眼科中心,510060 [2]中山大学基础医学院生物化学教研室

出　　处：《医学研究杂志》2006年第8期36-38,共3页Journal of Medical Research

基　　金：高校博士点专项科研基金(20040558076)资助

摘　　要：目的制备特异性的可溶性抗乳铁蛋白单链抗体。方法用固相化的乳铁蛋白从天然噬菌体抗体库筛选出抗乳铁蛋白的单链抗体克隆,转化至大肠杆菌HB2151中进行可溶性表达。表达产物以镍离子亲和层析法纯化,并进一步鉴定其纯度、活性等。结果成功筛选并表达了抗乳铁蛋白的单链抗体,该抗体能与乳铁蛋白特异性结合,而与转铁蛋白、溶菌酶、小牛血清白蛋白无交叉反应。结论建立了一种抗乳铁蛋白单链抗体的有效制备途径,为其进一步临床应用的研究奠定了基础。Objective To prepare a specific anti - lactoferrin single chain variable fragment （ScFv）. Methods Anti - lactoferrin clones were screened from a ＇ naive ＇ phage antibody library against the immobilized lactoferrin antigen, then the clones were transformed to the E. coli HB2151 to give soluble expression of antibody fragments. The culture supernatant containing ScFv was purified by immobilized metal affinity chromatography, and then determined with SDS - PAGE and ELISA. Results The results demonstrated that ScFvs were specific ; they did not react with transferrin, lysozyme and bovine serum albumin in ELISA. The SDS - PAGE showed that the ScFvs had high purity through affinity chromatography and the molecular weight of them was about 32 kD. Conclusions The successful generation of the ScFvs against lactoferrin provides a basis for further study and clinical applications for dry eye and other ocular diseases.

关 键 词：乳铁蛋白 噬菌体抗体库 单链抗体 筛选 

分 类 号：R151.2[医药卫生—营养与食品卫生学] R392.11[医药卫生—公共卫生与预防医学]