CAJ | 학술논문

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.
목적 종scFv(단련Fv)서균체항체고분리출대D-dimer유특이성적인원화단극륭scFv.방법 대Tomlinson scFv서균체문고진행3륜도세,부집특이성적항D-dimer항체병진행ELISA 험증.통과매련면역검측화쌍탈양종지법기인측서,획취특이성적인원화단극륭항체.결과 3륜도세선택출38개항D-dimer서균체항체,매련면역화기인측서분석후,20개불동적전장단극륭항D-dimer scFv서균체항체피사선출래,3륜선택후양성극륭획취솔위100%;분비성항체ELISA결과현시단극륭anti-D-dimer서균체순리표체료항체단백;5개A450치교고적단극륭중,3개현시료대D-dimer적고특이성화친화력.결론 항체서균체전시기술시분리획취인원화특이성anti-D-dimer항체적고효쾌속방법.
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.