从随机肽库中筛选抗人ANG抗体的模拟小肽

Screen of anti-angiogenin peptide from a phage-displayed random peptides library

目的从随机噬菌体十二肽库中寻找抗重组人 ANG抗体的模拟肽。方法以重组人 ANG为目标分子 ,对随机十二肽库进行亲合筛选 ,用噬菌体 EL ISA及竞争抑制实验鉴定阳性克隆的结合性和特异性。结果经过 3轮亲和筛选 ,噬菌体的回收率从 2 .0× 10 - 4 %增加到 9.3× 10 - 2 % ,阳性克隆得到富集 ;EL ISA和竞争抑制实验证明 :有 9个噬菌体克隆与靶分子有较强的结合活性 ,其中 6个阳性噬菌体克隆能特异性地抑制抗人重组 ANG单抗与 ANG的结合。结论该小肽可能是抗人ANG抗体的模拟小肽 ,可作为

ObjectiveTo isolate a peptide antagonist of the recombinant human angiogenin (rhANG) Methods A random peptide library of 12 amino acid residues displayed as a fusion to protein Ⅲ of filamentous phage fd Affinity selective methods were used to screen small peptides that bind to rhANG And the positive phage clones were identified with the methods of phage ELISA and competitive inhibition test Results After 3 rounds of screening, the gradual increase of the ratio of output to input were from 2 0×10 -4 % to 9 3×10 -2 %, which means that specific enrichment had been achieved Nine phage clones were identified as positive by phage ELISA and 6 of them could inhibit the interaction of rhANG with its antibody specifically by competitive binding ELISA Conclusion The peptide could act as potent mimetics of the antibody of anti rhANG, and could be used in a clinical medicine