利用噬菌体肽库筛选与HIV-1p24抗原结合的多肽

Screening of HIV-1 p24 Binding Peptides from Phage Display Peptide Library

通过噬菌体展示技术筛选出与HIV-1p24抗原结合的多肽,为用多肽辅助p24抗原检测提供实验基础。以重组p24抗原为靶蛋白,对噬菌体随机七肽库进行三轮筛选,用ELISA鉴定第三轮筛选到的噬菌体克隆与p24重组抗原的结合能力,再对噬菌体克隆进行测序分析,同时研究了ELISA中噬菌体加入量及多种封闭剂对噬菌体特异性结合能力的影响。在10个ELISA鉴定为阳性的噬菌体克隆中,有9个噬菌体克隆展示同一多肽序列FTTRANA,ELISA鉴定阳性克隆时噬菌体的加入范围为每孔8.13×1011～1.02×1014pfu,BSA作为封闭剂时效果最好。最终得到一种可与p24重组抗原结合的多肽,为多肽用于p24抗原检测打下基础。

A phage display peptide library was screened by targeting recombinant p24 antigen.After three rounds of panning,eleven phage clones were chosen to test the binding affinity by ELISA,and ten of them were positive.DNA sequencing showed that nine of the positive clones display a unique sequence FTTRANA which has high affinity with HIV-1 p24 antigen.To improve the condition of phage ELISA,phage number and blocking buffer have been optimized,phage number ranged from 8.13×1011 to 1.02×1014 pfu/well and 0.5%BSA was highly recommended.Therefore,in this study,peptides combined with p24 antigen were obtained which might be useful in p24 antigen assay.