ICAM-1特异结合肽的淘选与性质研究

Selection of the Peptides Binding to ICAM-1 from Phage Display Random 15mer Peptide Library

利用噬菌体展示载体pCANTAB5G8构建了随机十五肽库 ,根据转化率计算库容量为 1 36× 10 8。用PCR SSCP检验了库的异质性。以固相包被亲和淘选的方法 ,经过 4轮淘选和ELISA分析 ,得到了 5个可与重组人细胞间粘附分子 1(ICAM 1)特异结合的噬菌体克隆 ,对所展示的十五肽进行了序列分析。其中活性较高的噬菌体展示十五肽与ICAM 1的亲和常数约为 7 87× 10 7。

A phage display 15mer random peptide library expressed on the N\|terminal of the filamentous phage M13 minor coat protein pⅢ was constructed using the phagmid pCANTAB5G8.The capacity of library is 1 36×10\+8,calculated from the transformation efficiency.Human recombinant Intercellular Adhesion Molecule 1 was immobilized on the ELISA plates.After 4 rounds affinity panning and ELISA analysis,5 positive phage clones that can bind to ICAM\|1 specifically were selected.These positive clones were sequenced and the amino acid sequence of 15mer were deduced.The Ka of one positive clone was 7 87×10\+7,measured by ELISA.