一种用于抗原-抗体相互作用研究的实验方法  

An Experimental Method for the Study of Antigen-Antibody Interactions

作　　者：罗婷 刘美君 Daniel M.Czajkowsky Luo Ting;Liu Meijun;Daniel M.Czajkowsky(School of Biomedical Engineering,Shanghai Jiao Tong University,Shanghai,200240)

机构地区：[1]上海交通大学生物医学工程学院,上海200240

出　　处：《基因组学与应用生物学》2021年第2期849-854,共6页Genomics and Applied Biology

基　　金：国家自然科学基金项目(11374207,31670722,81627801)资助。

摘　　要：蛋白抗原-抗体相互作用是生物技术领域的重要研究之一。蛋白抗原难以合成、纯化且易于降解,低质量的蛋白抗原可能会导致该研究复杂、耗时、昂贵,甚至不可靠;抗原-抗体相互作用可能涉及易接触的抗原结合位点也可能给其带来挑战。故本研究开发出高效的实验体系,仅使用相应肽段作为抗原,来研究蛋白抗原与抗体的相互作用。以抗A型肉毒杆菌抗体及其对应的抗原肽段为模型,采用原子力显微镜和凝胶电泳确定研究的最佳条件。但抗原多肽在一些常规的缓冲液中会自发聚集。在确定了阻止其聚集的条件后,通过磁性微球分选洗脱实验模拟细胞表面的抗原-抗体结合过程,验证了该研究体系的有效性。故本研究所建立的实验体系高效、简单又经济,有望为免疫学基础研究、疫苗研发和免疫治疗等医学临床与工业应用发展提供技术补充。The interaction between protein antigens and antibodies is one of the most important studies in the biotechnol-ogical field.However,protein antigens are often difficult to be synthesized and purified,but easily degraded.Low-quality protein antigens may make the research complicated,time-consuming,expensive,and even unreliable;antigen-antibody interactions may involve easily accessible antigen binding sites,which may also pose challenges.Therefore,this study developed a highly efficient system to study protein antigen-antibody interactions that involves using unique peptide fragments from the protein instead of the whole protein as the antigen.Taking the anti-clostridium botulinum A toxin antibody and its corresponding antigenic peptide fragment as a model,we used atomic force microscopy and gel electrophoresis to identify optimal conditions to study this interaction.Surprisingly,we found that this peptide readily aggregated under a range of common buffer conditions,which completely prevented studies with antibodies.After identifying conditions that prevented this aggregation,we employed magnetic beads in a pull-down assay to examine the strength of this interaction,which we demonstrated was highly effective.Thus,the system we have developed,with particular attention to the buffer conditions,is an efficient,simple and cost-effective assay to study antigen-antibody interactions,which we expect will prove to be to be of use for basic immunology research as well as clinical applications,such as vaccine development and immunotherapy.

关 键 词：抗原-抗体反应 抗原多肽 聚集 原子力显微镜 

分 类 号：R392-3[医药卫生—免疫学]