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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。
The phage cyclic heptapeptide library was used to screen the polypeptides that specifically bind to hepatoma cells and their biological affinity was identified. HepG2 cells were screened by HL-7702 and HepG2 cells were screened by four rounds of cell subtraction, and 60 positive phage clones were randomly selected. The binding activity of HepG2 to HepG2 cells was determined by ELISA , And positive clones were sequenced and the synthesized peptides were identified by immunocytochemical staining. After 4 rounds of screening, phage displayed obvious enrichment on target HepG2 cells. Fifteen positive clones with high binding affinity to hepatocarcinoma cells were obtained by ELISA from randomly selected 60 phage clones. Sequencing analysis showed that amino acids Sequence homology, identified by immunocytochemical staining and found that a polypeptide sequence with high affinity, in order to improve antimicrobial peptides on tumor cells targeted killing basis.