用不同方法从大容量噬菌体抗体库中筛选抗完整食管癌细胞的单链抗体

Screening scFv of anti-intact carcinoma cells of esophageal squamous cell carcinoma from large phage library with different methods

目的:初步探索从天然的单链大容量噬菌体抗体库中筛选肿瘤细胞抗体的技术策略。方法:通过两种方法,即2%多聚甲醛固定细胞筛选法(PF)及活细胞直接筛选法(NF),经过4轮"吸附-洗脱-扩增"的淘筛过程,对食管癌KYSE-170细胞进行筛选,得到抗食管癌细胞的噬菌体抗体,完善了噬菌体抗体库筛选完整细胞的特异性抗体的技术策略。并且进一步制备了其可溶性抗体。结果:活细胞NF组的筛选阳性率高于2%多聚甲醛固定PF组,筛选特异性抗体的阳性率分别为25.00%和12.50%,两组的差异具有统计学意义。结论:利用单链大容量抗体库可以采用不同的筛选方法成功制备与肿瘤细胞结合的噬菌体抗体,为今后的研究和应用奠定基础。

ObjectiveTo establish the method for human anti - tumor cells antibodies from large phage antibody library. Methods ：Through two methods,2% paraformaldehyde fixed cell screening method （ PF） and living cells di-rect screening method （NF） ,after four rounds of ＂ adhesion,elution and amplification＂ process,positive phage anti-bodies were identified for esophageal KYSE - 170 cells. It was a technology strategy for screening specific cell anti-body from the phage antibody library. Results ： The positive screening rate of NF by cell ELISA was higher than PF （25.00% vs 12. 50% ） and soluble scFv was produced. The significant difference between the two groups had statisti-cal significance （P 〈0. 05） . Conclusion： The large phage library biopanning was advantageous in generating specific scFv antibodies for cells by different methods.