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目的应用噬菌体随机七肽库生物淘选人角质细胞生长因子(keratinocyte growth factor,KGF)噬菌体模拟肽。方法以人KGF单克隆抗体为靶,对噬菌体随机七肽库进行4轮生物淘选,测定噬菌体滴度。随机挑取单克隆噬菌体行ELISA检测其结合力,对结合活性较好的单克隆噬菌体提取噬菌体DNA,测序,并应用Basic BLAST系统行序列相似性和同源性分析。结果经过4轮生物淘选获得的噬菌体滴度逐渐升高,特异性噬菌体模拟肽获得了富集,行ELISA检测的单克隆噬菌体滴度均可达到2.0×1014 pfu/mL。取吸光度值显示与特异性抗体具有较好结合性的单克隆噬菌体进行测序,共获得26个与促进分裂、生长有关的碱基序列,其中有2个序列与人KGF相似;同源序列分析显示26个碱基序列的共同序列与人KGF的部分序列相似。结论从噬菌体随机七肽库中可淘选到与人KGF DNA序列相似或相关的噬菌体模拟肽,有望改善人KGF性能,促进创面愈合和改善组织工程皮肤替代物的性能。
OBJECTIVE: To screen the phage display peptide of human keratinocyte growth factor (KGF) by phage random peptide library. Methods Human KGF monoclonal antibody was used as a target to perform 4 rounds of biopanning on phage random peptide library to determine phage titer. The phage DNA of the monoclonal phage with good binding activity was extracted and sequenced. The similarity and homology of the sequences were analyzed by Basic BLAST system. Results After 4 rounds of biopanning, the titer of phage was gradually increased and the specific phage peptidomimetic peptide was enriched. The titer of monoclonal phage was 2.0 × 1014 pfu / mL. A total of 26 single-phage clones with good binding ability to specific antibodies were sequenced and 26 base pairs were found to be involved in promoting division and growth. Two of them were similar to human KGF. Homology analysis The consensus sequence of 26 base sequences is similar to the partial sequence of human KGF. CONCLUSION: The phage peptidomimetics, which are similar to or related to human KGF DNA sequence, can be panned out from the phage random peptide library. It is expected to improve the performance of human KGF, promote wound healing and improve the performance of tissue engineered skin substitutes.