【作者】 郭莉娜；

【导师】 韩继媛；

【作者基本信息】 华中科技大学 ， 急诊医学， 2010， 硕士

【摘要】 目的从大型人源性噬菌体抗体库中直接筛选出百草枯特异性单链抗体(scFv),并与百草枯人工抗原特异性结合性能进行鉴定。方法以百草枯人工抗原(PQ-h-BSA)为靶抗原,采用固相化抗原免疫吸附筛选法,通过“吸附-洗脱-扩增”筛选富集过程,从大型人源性噬菌体抗体库中筛选出特异性百草枯单链抗体,通过ELISA法对其特异性结合性能进行初步鉴定。结果经过5轮筛选,获得两株能与百草枯人工抗原特异性结合的阳性克隆。经ELISA法初步鉴定,筛选出的单链抗体与百草枯人工抗原反应强烈,而与其他抗原不反应或反应微弱。结论利用噬菌体抗体技术可不经免疫制备人源性百草枯抗体,为临床治疗百草枯中毒研究提供了具有更广阔前景的人源性小分子抗体。更多还原

【Abstract】 Objective To screen humanized anti-paraquat scFv from large human phage antibody library ,and its specific binding properties with paraquat artifical antigen were identified.Methods In the process of "absorption - elution - amplification" , using solid-phase ELISA-based antigen screening method, that artificial antigen paraquat (PQ-h-BSA) as antigen,choosing by their specific binding properties of the initial identification in ELISA, to screen humanized anti-paraquat scFv from large human phage antibody library .Results Two strains with paraquat artificial antigen-specific combination of positive clones were got in 5 rounds of screening. In initially identified by ELISA, anti-paraquat scFv of screeninig reacted strongly with paraquat artifical antigen , but no reaction or weak with other antigens.Conclusion It is feasible to obtain humanized anti-paraquat scFv by phage display technique,which provides a more promising source of small molecule human antibodies for clinical treatment of paraquat poisoning study.更多还原