从人天然抗体库中克隆IgM抗角蛋白抗体
ZGM,,ZGM;噬菌体;抗体,细菌;角蛋白,1材料和方法,2结果,参考文献
关键词: ZGM;噬菌体;抗体,细菌;角蛋白中图号:R392.11
摘 要:目的 克隆制备人源性抗角蛋白IgM抗体. 方法 以人表皮角蛋白为抗原,从人源性天然抗体库中筛选IgM型抗角蛋白人抗体,并用ELISA对所获抗体进行抗原结合活性和特异性等鉴定. 结果 通过3轮“亲和吸附-洗脱-扩增”筛选,获得2株IgM型抗角蛋白噬菌体抗体.酶切去掉载体上的噬菌体基因Ⅲ,得到了可溶性表达的Fab段.经过鉴定,所获抗体具有良好的抗原结合活性和特异性. 结论 从人源性天然抗体库中成功筛选出2株抗角蛋白IgM抗体,为进一步研究抗角蛋白抗体的特性提供了条件.
Cloning of anti┐keratin IgM antibodies from a hu┐man natural antibody library
WANG Gang,LIU Yu-Feng,WANG Yan
Center of Dermatology&Venereology of Chinese PLA,Xijing Hospital,Fourth Military Medical U-niversity,Xi'an710033,China,Department of Central Experiment,Navy General Hospital,Bei-jing100037,China
Keywords:ZGM;bacteriophages;antibodies,bacterial;ker-atin
Abstract:AIM To clone human anti-keratin IgM antibodies from a natural antibody library.METHODS Panning of the natural antibody library against keratin antigen was conduct-ed to select specific IgM antibodies.The antigen binding ac-tivity and the specificity of the isolated binders were deter-mined with ELISA.RESULTS After3rounds of“adhen-sion-amplification-elution”panning,2clones of IgM which could bind specifically to keratin were obtained.After remov-ing the phagesome gene III fragment ......
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