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目的全细胞差减法筛选人源性卵巢癌HO8910细胞株特异性结合短肽,为卵巢癌的靶向药物治疗提供理想的载体。方法以人卵巢癌HO8910细胞为靶细胞,人正常卵巢上皮细胞为吸附细胞,对噬菌体随机环7肽库进行4轮差减筛选;ELISA验证阳性噬菌体克隆;对获得的阳性克隆进行DNA测序及生物信息学分析,采用免疫荧光细胞化学方法鉴定噬菌体阳性克隆(phage1)与HO8910细胞的特异性结合。结果经过4轮筛选后,噬菌体在靶细胞HO8910上出现了明显的富集现象;ELISA对20个随机挑选的克隆进行鉴定,12个可与HO8910特异性结合;DNA测序后得到一段与卵巢癌细胞特异性结合的7肽(SWQIGGN),免疫荧光细胞化学结果显示phage1能特异性结合HO8910细胞。结论采用全细胞差减法成功筛选到与卵巢癌细胞特异性结合的7肽。
OBJECTIVE: To screen the HO8910 cell line specific binding short peptide by whole cell subtractive subtraction to provide an ideal carrier for targeted drug therapy of ovarian cancer. Methods Human ovarian cancer HO8910 cells were used as target cells and normal human ovarian epithelial cells were used as adherent cells. The phage random loop 7-peptide library was screened by four rounds. The positive clones were verified by ELISA. The positive clones were sequenced and sequenced. Informatics analysis identified the specific binding of phage1 to HO8910 cells by immunofluorescence cytochemistry. Results After 4 rounds of screening, the bacteriophage showed obvious enrichment on target cell HO8910. Twenty randomly selected clones were identified by ELISA and 12 were specifically combined with HO8910. After DNA sequencing, Specific binding of the peptide 7 (SWQIGGN), immunofluorescence cytochemistry results show that phage1 can specifically bind HO8910 cells. Conclusion We successfully screened the 7 peptide which is specific to ovarian cancer cells by whole cell subtraction method.