肝癌细胞靶向肽的筛选与鉴定  

Screening and characterization of multi-peptide binding specifically to hepatocellular carcinoma cells by phage peptide library

作　　者：赵红蕾[1] 韩文瑜[1] 刁昱文[1] 冯新[1] 韩东[1] 朱僧[1] 刘晓贺[1] 路荣[1] 雷连成[1] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062

出　　处：《中国兽医学报》2012年第2期202-206,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30901069);国家自然科学基金重点资助项目(31130052)

摘　　要：利用噬菌体环七肽库筛选与肝癌细胞特异性结合的多肽,并对其亲和力进行生物学鉴定。以HL-7702为消减细胞,HepG2为筛选靶细胞,对噬菌体随机环七肽库进行4轮全细胞消减筛选,并随机挑取60个阳性噬菌体克隆,以ELISA法鉴定其与HepG2细胞的结合活性,并取阳性克隆进行测序分析,并合成多肽进行免疫细胞化学染色鉴定。经4轮筛选,噬菌体在靶细胞HepG2上出现明显富集;利用ELISA从随机挑选的60个噬菌体克隆中得到15个与肝癌细胞具有高结合力的阳性克隆,测序并进行序列分析比对发现氨基酸序列无同源性,经免疫细胞化学染色鉴定后,发现1条多肽序列亲和力较高,为提高抗菌肽对肿瘤细胞的靶向杀伤作用奠定基础。Ph.D-C7CTM peptide libraries were used to screen and characterize peptides which bind specifically to hepatocellular carcinoma cells.Whole-cell subtractive screening was performed on HepG2 cells and HL-7702 cells,in which hepatocellular carcinoma cells were used as the target cells and normal cells as the absorber cells.The positive phage clones selectived stochastically were identified by using cell-based ELISA and analyzed by DNA sequencing,following with immunocytochemistry.After 4 rounds of panning,phages which can bind to the HepG2 cells were enriched obviously.Fifteen positive clones with high affinity were identified by ELISA.After cell immunofluorescence and immunocytochemistry,we found 1 peptide with high affinity and no conserved motif was found in these peptides,which increase the targeting of antimicrobial peptides on tumor cells.

关 键 词：噬菌体随机环七肽库 肝癌细胞 消减筛选 靶向 

分 类 号：S859[农业科学—临床兽医学]