肝癌细胞靶向肽的筛选与鉴定 |
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引用本文: | 赵红蕾,韩文瑜,刁昱文,冯新,韩东,朱僧,刘晓贺,路荣,雷连成.肝癌细胞靶向肽的筛选与鉴定[J].兽医大学学报,2012(2):202-206. |
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作者姓名: | 赵红蕾 韩文瑜 刁昱文 冯新 韩东 朱僧 刘晓贺 路荣 雷连成 |
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作者单位: | 吉林大学畜牧兽医学院,吉林长春130062 |
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基金项目: | 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) |
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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。
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关 键 词: | 噬菌体随机环七肽库 肝癌细胞 消减筛选 靶向 |
Screening and characterization of multi-peptide binding specifically to hepatocellular carcinoma cells by phage peptide library |
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Authors: | ZHAO Hong-lei HAN Wen-yu DIAO Yu-wen FENG Xin HAN Dong ZHU Seng LIU Xiao-he LU Rong LEI Lian-cheng |
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Institution: | (College of Animal Science and Veterinary Medicine,Jilin University,Changchun 130062,China) |
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Abstract: | 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. |
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Keywords: | Ph D-C7CTM peptide library hepatocellular carcinoma cells substaction biopanning targeting |
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