CAJ | 학술논문

目的从scFv噬菌体库中获取人源化特异性Anti-GPⅡb/Ⅲa单克隆scFv抗体.方法对Tomlinson I+J scFv文库进行3次淘洗,富集特异性的抗GPⅡb/Ⅲa抗体.通过酶联免疫吸附试验(ELISA)和双脱氧终止法基因测序及同源性对比,检测出人源化的抗GPⅡb/Ⅲa单克隆抗体.结果　在3次淘洗后,得到了抗GPⅡb/Ⅲa单克隆噬菌体抗体,阳性克隆的获取率在95.6%以上;ELISA和基因测序筛选出25种不同的全长抗GPⅡb/Ⅲa噬菌体抗体,这些基因序列与人免疫球蛋白可变区基因同源性达到89%以上;分泌性抗体ELISA检测提示这些抗体顺利表达了蛋白并特异性结合GPⅡb/Ⅲa,其中15种scFv对GPⅡb/Ⅲa有更强的阳性反应.结论　人源化的特异性抗-GPⅡb/Ⅲa scFv能通过噬菌体展示技术快速有效地获得.
목적 종scFv서균체고중획취인원화특이성Anti-GPⅡb/Ⅲa단극륭scFv항체.방법 대Tomlinson I+J scFv문고진행3차도세,부집특이성적항GPⅡb/Ⅲa항체.통과매련면역흡부시험(ELISA)화쌍탈양종지법기인측서급동원성대비,검측출인원화적항GPⅡb/Ⅲa단극륭항체.결과　재3차도세후,득도료항GPⅡb/Ⅲa단극륭서균체항체,양성극륭적획취솔재95.6%이상;ELISA화기인측서사선출25충불동적전장항GPⅡb/Ⅲa서균체항체,저사기인서렬여인면역구단백가변구기인동원성체도89%이상;분비성항체ELISA검측제시저사항체순리표체료단백병특이성결합GPⅡb/Ⅲa,기중15충scFv대GPⅡb/Ⅲa유경강적양성반응.결론　인원화적특이성항-GPⅡb/Ⅲa scFv능통과서균체전시기술쾌속유효지획득.
Objective To screen monoclonal anti-GP Ⅱ b/Ⅲ a antibodies from scFv phage libraries and obtain specific monoclonal anti-GP Ⅱ b/Ⅲ a scFv. Methods Specific anti-GP Ⅱ b/Ⅲ a scFv antibodies were enriched by three rounds of selection from Tomlinson Ⅰ + J libraries. By using polyclonal and monoclonal phage enzyme linked immunosorbent assay ( ELISA) and gene sequencing by bideoxy chain termination, full-length specific monoclonal anti-GP Ⅱ b/Ⅲ a antibodies were picked out and their gene sequences were obtained. Results Twenty-five different full-length monoclonal scFv phage fragments were obtained after three rounds of panning and their gene sequences were identified, and positive rate of monoclones was above 95.6%; homology comparison with variable regions of human immunoglobulin gene showed the similarity was above 89%; the result of soluble scFv ELISA showed that these specific scFv could be expressed smoothly, and 15 full-length monoclonal scFv antibodies were stronger positive than the other in these scFv. Conclusion Antibody phage display was a rapid and effective method to obtain Anti-GP Ⅱ b/Ⅲ a scFv fragements.