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Screening and Identification of a peptide specifically targeted to NCI-H1299 from a phage display peptide library
Authors:Linquan Zang  Lei Shi  Jiao Guo  Qin Pan  Wei Wu  Xuediao Pan  Junye Wang
Institution:1. Department of Pharmacology, Novel Drug Screening Center, School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006, Guangdong, PR China;2. The Institute of Traditional Chinese Medicine of Science, Guangdong Pharmaceutical University, Guangzhou 510006, Guangdong, PR China;3. Sun Yat-sen University Cancer Center, Guangzhou 510060, Guangdong, PR China
Abstract:In this study, a NCI-H1299 (Non-Small Cell Lung Cancer, NSCLC) and a normal lung cell line (Small Airway Epithelial Cells, SAEC) were used for the subtractive screening in vitro with a phage display-12 peptide library. After three rounds of panning, there was an obvious enrichment for the phages specifically binding to the NCI-H1299 cells, and the output/input ratio of phages increased about 875-fold (from 0.4 × 104 to 3.5 × 106). A group of peptides being capable of binding specifically to the NCI-H1299 cells were obtained, and the affinity of these peptides to bind to the targeted cells and tissues was studied. Through a cell-based ELISA, immunocytochemical staining, immunohistochemical staining, and immunofluorescence, a M13 phage isolated and identified from the above screenings, and a synthetic peptide ZS-1 (sequence EHMALTYPFRPP) corresponded to the sequence of the surface protein of the M13 phage were demonstrated to be capable of binding to the tumor cell surfaces of NCI-H1299 and A549 cell lines and biopsy specimens, but not to normal lungs tissue samples, other different cancer cells, or nontumor surrounding lung tissues. In conclusion, the peptide ZS-1 may be a potential candidate of biomarker ligands used for targeted drug delivery in therapy of lung cancer.
Keywords:Lung cancer  Peptide  Phage display  Biomarker  NCI-H1299  Phage M13
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