In vitro panning of a targeting peptide to hepatocarcinoma from a phage display peptide library
ABSTRACT Phage display technology has been used as a powerful tool in the discovery of ligands specific to receptor(s) on the surface of a cancer cell and could also impact clinical issues including functional diagnosis and cell-specific drug delivery. After three rounds of in vitro panning and two rounds of reverse absorption, a group of phages capable of addressing BEL-7402 enormously were obtained for further analysis. Through a cell-based ELISA, immunofluorescence, FACS, and in vivo binding study, WP05 (sequence TACHQHVRMVRP) was demonstrated to be the most effective peptide in targeting four kinds of liver cancer cell lines (BEL-7402, BEL-7404, SMMC-7721, and HepG2), but not the normal liver cell line HL-7702. In conclusion, the peptide WP05 which was screened by in vitro phage display technology was proved to be a targeting peptide to several common hepatocellular carcinoma cell lines.
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ABSTRACT: Anchored periplasmic expression (APEx) is a method for isolating high affinity ligand-binding proteins from large combinatorial libraries, and antibodies highly specific for soluble antigens were successfully isolated from APEx antibody libraries in combination with flow cytometric sorting (Harvey et al., Proc Natl Acad Sci USA 101(25):9193-9198, 2004). However, many disease markers and drug targets are localized on the cell surface, and often, unique posttranslational modifications and/or properly folded epitopes are lost when they were expressed and isolated in soluble form. In this study, we demonstrate that Escherichia coli spheroplasts, displaying antibodies and screened by a combination of plate-panning and flow cytometric sorting, can be used for isolating antibodies specific for antigens on the human cell surface. Two rounds of plate-panning followed by one round of flow cytometric sorting resulted in 7,200-fold enrichment of antibodies specific for the protective antigen of Bacillus anthracis from a large excess of spheroplasts expressing a scFv antibody to digoxin (a negative control). There is the potential to use this technique for library screening to find novel antibodies against disease cell surface antigens.Applied Microbiology and Biotechnology 09/2010; 88(6):1385-91. DOI:10.1007/s00253-010-2861-3 · 3.81 Impact Factor
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ABSTRACT: ObjectiveTo identify and localize the synthesized targeting peptide A54 to liver cancer cell line BEL-7502 in vivo and in vitro for confirming the potential clinical application of peptide A54 in hepatocarcinoma targeting therapy. MethodsPhage A54 was confirmed by ELISA. Biotin and FAM labeled A54 peptides were identified and localized by means of immunohistochemistry and immunocytochemistry. ResultsA54 peptide could target the liver-tumor tissue in vivo and adhere to several liver-tumor cells in vitro. FAM-labeled A54 peptides were localized on the membrane surface of liver-tumor cells. ConclusionSynthesized A54 peptide obtained from in vivo phage display technology still kept special ability to adhere liver-tumor cell in vivo and in vitro. The A54 peptide could be a candidate carrier for hepatocarcinoma targeting therapy.Chinese Journal of Cancer Research 11/2006; 18(4):241-245. DOI:10.1007/s11670-006-0241-4 · 0.93 Impact Factor
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ABSTRACT: Combinatorial peptide library technology is a valuable resource for drug discovery and development. Several peptide drugs developed through phage-displayed peptide library technology are presently in clinical trials and the authors envision that phage-displayed peptide library technology will assist in the discovery and development of many more. This review attempts to compile and summarize recent literature on targeting peptides developed through peptide library technology, with special emphasis on novel peptides with targeting capacity evaluated in vivo.Expert Opinion on Drug Discovery 04/2007; 2(4):525. DOI:10.1517/174604188.8.131.525 · 3.47 Impact Factor