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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 ANG的拮抗剂
Objective To search for mimetic peptides against recombinant human ANG antibodies from a random phage dideopeptide library. Methods Recombinant human ANG was used as the target molecule to screen the randomized 12-mer peptide library. The binding and specificity of the positive clones were confirmed by phage ELISA and competitive inhibition assay. Results After 3 rounds of affinity screening, the phage recoveries increased from 2.0 × 10 - 4% to 9.3 × 10 - 2%, and the positive clones were enriched. The EL ISA and competitive inhibition experiments showed that there were 9 phage clones with The target molecules have strong binding activity, of which 6 positive phage clones can specifically inhibit the binding of anti-human recombinant ANG MAb to ANG. Conclusion The small peptide may be an anti-human ANG antibody mimic peptide, which can be used as an antagonist of ANG