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目的从噬菌体展示文库中筛选能与抗人PTA1单克隆抗体(mAbs)特异性结合的短肽。方法采用protein-A亲和层析法纯化系列抗人PTA1mAbs，通过流式细胞术检测筛选出能够阻断PTA1-Fc融合蛋白与其天然配体结合的mAb，经过三轮亲和筛选，从噬菌体随机12肽库中筛选可特异性结合mAb的重组噬菌体阳性克隆，进而对这些克隆的短肽序列进行测定和分析。结果确定了能够阻断PTA1-Fc融合蛋白与其天然配体结合的mAb为LeoA1，得到了13个具有特异性结合LeoA1的重组噬菌体阳性克隆，序列测定结果发现，这些可结合LeoA1的短肽，有较保守而集中的模式序列，即WPXHHX序列。结论这些具有保守模式序列的短肽，有可能模拟PTA1分子的功能表位，成为其天然配体拮抗剂的候选肽段。此外，PTA1分子与其配体结合的功能表位的确定，也为今后寻找天然配体和进一步深入研究PTA1分子的结构和功能提供了实验依据。
Objective To screen short phage display libraries for specific binding to anti-human PTA1 monoclonal antibodies (mAbs). Methods A series of anti-human PTA1 mAbs was purified by protein-A affinity chromatography. The mAbs that could block the binding of PTA1-Fc fusion protein with its natural ligand were screened by flow cytometry. After three rounds of affinity screening, 12 peptide library, recombinant phage-positive clones that specifically bind to the mAb were screened, and the short peptide sequences of these clones were determined and analyzed. As a result, it was confirmed that the mAb capable of blocking the binding of the PTA1-Fc fusion protein to its natural ligand was LeoA1, and 13 recombinant phage-positive clones that specifically bound to LeoA1 were obtained. As a result of sequencing, these short peptides binding to LeoA1 The more conservative and focused pattern sequence, the WPXHHX sequence. Conclusions These short peptides with conserved pattern sequences are likely to mimic the functional epitopes of PTA1 molecules and become candidate peptides for their natural ligand antagonists. In addition, the determination of the functional epitope of PTA1 binding to its ligand also provides experimental evidence for searching for natural ligands and further studying the structure and function of PTA1 molecules in the future.