应用噬菌体肽库筛选Endoglin的结合肽

目的 利用噬菌体12肽库筛选可与Endoglin结合的活性小分子肽.方法 以rhEndoglin为靶分子,对噬菌体12肽库进行亲和筛选.经过3轮筛选,随机挑取16个克隆,双夹心ELISA法鉴定其亲和性,测定亲和力高的阳性噬菌体克隆的DNA序列,推断出与噬菌体外壳蛋白融合的氨基酸序列,竞争抑制实验检测优势克隆的特异性.结果 竞争性ELISA显示6个噬菌体克隆与rhEndoglin有较强的结合活性,经测序获得5种不同的多肽序列,其中2个克隆的氨基酸序列为:AHKHVHHVPVRL.结论 噬菌体随机肽库技术可以获得Endoglin结合肽的蛋白质序列.

Screening of conjugated peptides of Endoglin from phage display peptide library

Objective To screen rhEndoglin-binding peptides from phage displayed 12-peptide library. Methods The rhEndoglin was used as target protein for biopanning of phage-displayed 12-peptide library. After three rounds of screening, 16 phage clones were randomly selected and identified by sandwich ELISA. The positive phage clones were sequenced, and the fuse peptides were deduced by the DNA sequence. Further we identified the affinity and speciality by competitive inhibition test. Results Six of 16 phage clones were identified as positive clones by competent ELISA which could bind to rhEndoglin. Five sequences were obtained, and the amino acid sequence in two of these five was AHKHVHHVPVRL. Conclusion The rhEndoglin-binding peptides can be obtained by screening phage random peptide library. It could play an important role in early diagnosis of ovarian cancer.