摘要
目的:利用合成的重组Neuropilin-1的蛋白分子对噬菌体随机七肽库进行筛选,以期获得能与Neuropilin-1分子具高亲和活性的小肽分子,并对其功能进行初步鉴定。方法:对包被好的rhNeuropi-lin-1蛋白,经过三轮"吸附-洗脱-扩增"后,ELISA鉴定阳性克隆,DNA测序和分析,竞争抑制试验进一步分析其相关化学合成多肽的功能。结果:经3轮亲和筛选后,噬菌体克隆得到有效富集,ELISA鉴定结果显示2个阳性克隆具有较强结合活性。DNA测序显示其具有*FPS***的一致基序。阳性克隆与Neuropilin-1分子的结合能被其相关化学合成多肽FITC-DFPSPLW所抑制。结论:利用噬菌体随机展示肽库技术获得了与Neuropilin-1分子具特异性结合能力的小肽分子,为进一步基于Neuropilin-1的肿瘤血管生成抑制的研究奠定基础。
Objective： To screen small peptides binding Neuropilin-1 in phage displayed 7-peptide library and to preliminarily identify its function. Methods. rhNeuropilin-1 protein as selected ligands under- went three rounds of ＂absorption-elution-amplification＂ process. Then they were identified by ELISA. DNA sequence and analysis were performed. Its function as chemical synthesis of polypeptide was analyzed by competitive inhibition test. Result. After 3 affinity screening, the phage clones were gathered. ELISA result revealed 5 of 12 phage clones were positive with a strong binding activity. And DNA sequence showed sequence motif same with ＊ FPS ＊＊ ~. Positive clone combined with rhNeuropilin-1 was inhibited by chemical synthesized peptides ＂FITC-DFPSPLW＂. Conclusion： The rhNeuropilin-l-biding peptides with high affinity to Neuropilin can be acquired by tphage displayed random peptide library, which provide basis for studies on the function of Neuropilin-based inhibitors for tumor angiogenesis.
出处
《海南医学院学报》
CAS
2013年第4期442-445,448,共5页
Journal of Hainan Medical University
基金
中国高校医学期刊临床专项资金项目(112210582)~~
