抗人凝血因子Ⅷ-C1C2区单克隆抗体的制备及鉴定  

Preparation and Characterization of Monoclonal Antibody to FⅧ C1C2 Domain of Human

作　　者：张婷婷[1] 赵益明[1] 李振宇[1] 沈飞[1] 阮长耿[1] 

机构地区：[1]苏州大学附属第一医院江苏省血液研究所卫生部血栓与止血重点实验室,江苏苏州215006

出　　处：《苏州大学学报（医学版）》2010年第1期106-109,共4页Suzhou University Journal of Medical Science

基　　金：江苏省高校自然科学基金资助项目(08KJD310008);江苏省卫生厅"科教兴卫工程"临床医学中心血液病开放课题基金资助项目(WKF07009)

摘　　要：目的制备抗人凝血因子Ⅷ(FⅧ)C1C2区单克隆抗体(mAb),并对其进行生化和生物学鉴定。方法采用基因重组技术表达FⅧ-C1C2区蛋白(rFⅧ-C1C2),以rFⅧ-C1C2免疫8周龄Balb/C小鼠,小鼠脾脏细胞与小鼠骨髓瘤细胞融合后,ELISA法筛选阳性克隆,制备稳定分泌抗rFⅧ-C1C2mAb的细胞株,并对mAb进行生化和免疫学鉴定。结果获得一株抗rFⅧ-C1C2区单克隆抗体阳性克隆2B11,命名为SZ-133。琼脂糖双扩散鉴定为IgG1类,Westernblot法证实SZ-133可与rFⅧ-C1C2区蛋白及重组的全长FⅧ特异性结合,但不影响FⅧ的凝血活性。结论表达了rFⅧ-C1C2,并制备出了抗人凝血因子Ⅷ的单克隆抗体SZ-133,它能特异识别血浆及重组的FⅧ,有可能用于基础和临床应用研究。Objective To develop monoclonal antibody（mAb）anti-FⅧ C1C2 domain of human and characterize its biochemical and biological functions.Methods FactorⅧ-C1C2 protein was expressed by prokaryotic expression system.Balb/c mice of eight-week ages were immunized by intraperitoneal and subcutaneous injection of rFⅧ-C1C2 protein.The positive clone was screened by ELISA and identified using western-blot.Results A positive clone 2B11 designated as SZ-133 was produced,belonging to IgG1 subclass.Western blot demonstrated that SZ-133 could specifically recognized rFⅧ-C1C2 and fⅧ in human plasma.Conclusion rFⅧ-C1C2 is expressed in Ecoli.The successfully developed mAb against human FⅧ-C1C2 domain mAb can be useful for basic and clinical application.

关 键 词：人凝血因子Ⅷ(FⅧ)C1C2区 原核表达 单克隆抗体 鉴定 

分 类 号：R331.1[医药卫生—人体生理学]