中文摘要：

目的研制抗rhEndoglin-scFv(single chain variable fragment)抗体。方法将具有抗原结合活性的抗rhEndoglin-scFv噬菌体感染大肠杆菌E.coliHB2151,经IPTG诱导表达,SDS-PAGE和Western Blot鉴定分析。用HiTrap Anti-ETag柱亲和层析纯化scFv抗体,HiTrap Desalting柱行缓冲液更换。用间接ELISA测scFv抗体的亲和常数,用竞争性ELISA和间接免疫荧光检测scFv抗体的抗原结合活性。结果成功诱导表达和纯化抗rhEndoglin-scFv抗体,表达产物主要位于周质腔中,相对分子质量约为2.9×104。测定scFv的亲和常数为(2.14±0.84)×107L/mol,它能与抗rhEndoglin单抗竞争性结合同一抗原表位,且竞争作用随scFv浓度增加而加强。间接免疫荧光实验证实该scFv抗体能识别HUVEC胞膜表达的Endoglin抗原。结论制备的抗rhEndoglin-scFv抗体具有与rhEndoglin和天然Endoglin结合的能力,可望将其用作肿瘤诊断和治疗的靶向载体。

英文摘要：

Objective To prepare scFv antibody against rhEndoglin(recombinant human Endoglin).Methods E.coli HB2151 was infected by phage displaying scFv fragment against rhEndoglin,and then induced by IPTG to express the soluble scFv antibody against rhEndoglin,which was identified and analyzed by SDS-PAGE and Western blot.Purification and buffer exchange of the scFv antibody were performed respectively with HiTrap Anti-E tag columns and HiTrap desalting columns.Affinity constant of the scFv antibody was determined by...