从scFv噬菌体库分离特异性的人源化抗D-dimer抗体  

Isolation of specific humanized anti-D-dimer scFv fragments from scFv phage libraries

作　　者：夏红利[1] 谭最[1] 陈德杰[2] 乔建国[1] 邱仁峰[1] 

机构地区：[1]武汉大学中南医院普外科,430071 [2]襄樊市中心医院普外科

出　　处：《中华微生物学和免疫学杂志》2011年第2期168-172,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目（30872525）

摘　　要：目的 从scFv（单链Fv）噬菌体抗体库分离出对D-dimer有特异性的人源化单克隆scFv.方法 对Tomlinson scFv噬菌体文库进行3轮淘洗,富集特异性的抗D-dimer抗体并进行ELISA 验证.通过酶联免疫检测和双脱氧终止法基因测序,获取特异性的人源化单克隆抗体.结果 3轮淘洗选择出38个抗D-dimer噬菌体抗体,酶联免疫和基因测序分析后,20个不同的全长单克隆抗D-dimer scFv噬菌体抗体被筛选出来,3轮选择后阳性克隆获取率为100% 分泌性抗体ELISA结果显示单克隆anti-D-dimer噬菌体顺利表达了抗体蛋白 5个A450值较高的单克隆中,3个显示了对D-dimer的高特异性和亲和力.结论 抗体噬菌体展示技术是分离获取人源化特异性anti-D-dimer抗体的高效快速方法.Objective To isolate specific humanized anti-D-dimer scFv（single chain Fv） antibody from scFv phage libraries. Methods Isolate anti-D-dimer positive clones from Tomlinson I ＋ J phage libraries by three rounds of panuing, then sequence monoclonal genes by bideoxy-mediated chain termination and express soluble scFv antibody Pick out anti-D-dimer antibodies with high specificity and affinity by ELISA.Results After three rounds of selection from human scFv phage libraries Tomlinson I and J, 38 monclonal specific anti-D-dimer scFv fragments were selected. By polyclonal and monoclonal phage ELISA and gene sequencing, 20 different full-length monoclonal scFv phages were identified, the result of soluble scFv ELISA showed that 20 full-length monoclonal scFv were expressed smoothly. According to the result of soluble scFv ELISA, in 5 scFv antibodies with high value of A450 selected, 3 scFv antibody fragments showed high specific and affinity. Conclusion Antibody phage display was an effective, rapid method to isolate anti-D-dimer antibodies with high specificity and affinity.

关 键 词：筛选 抗D-dimer抗体 噬菌体展示技术筛选 抗D-dimer抗体 噬菌体展示技术筛选 抗D-dimer抗体 噬菌体展示技术 

分 类 号：R392[医药卫生—免疫学]