Article

In vitro panning of a targeting peptide to hepatocarcinoma from a phage display peptide library.

School of Life Sciences, East China Normal University, Shanghai, PR China.
Biochemical and Biophysical Research Communications (Impact Factor: 2.28). 05/2006; 342(3):956-62. DOI: 10.1016/j.bbrc.2006.02.050
Source: PubMed

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.

1 Bookmark
 · 
104 Views
  • [Show abstract] [Hide abstract]
    ABSTRACT: Brucellosis is a globally distributed zoonotic disease that causes animal and human diseases, while current antibiotic and vaccine strategies are not optimal. The surface-exposed protein OMP25 is involved in Brucella virulence and play an important role in Brucella pathogenesis during infection, suggesting that Omp25 is an useful target for selecting potential therapeutic molecules to inhibit Brucella pathogenesis. In this study, we identified, for the first time, the specific peptides bind specifically to Omp25 protein of pathogen using a phage panning technique, After four rounds of panning, 42 plaques of eluted phages were subjected to pyrosequencing. Four phage clones bound better than the other clones were selected through the confirmation of ELISA and affinity constants. Selected peptides could inhibit significantly Brucella abortus 2308 (S2308) internalization and intracellular growth in RAW264.7 macrophages, and induce significantly secretion of tumor necrosis factor alpha (TNF-α) and interleukin-12 in peptide- and S2308-treated cells. Any observed peptide (OP11, OP27, OP35, or OP40) could inhibit significantly S2308 infection in BALB/c mice. Moreover, the peptide 0P11 was the best candidate peptide in inhibiting S2308 infection in vitro and in vivo. These results suggest that peptide OP11 has potential for exploitation as a peptide drug in resisting S2308 infection.
    Journal of Medical Microbiology 04/2014; · 2.30 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: There is an urgent need for biomarkers to identify malignant thyroid nodules from indeterminate follicular lesions. We have used a subtractive proteomic strategy to identify novel biomarkers by selecting ligands to goiter tissue from a 12-mer random peptide phage-displayed library using the BRASIL method (Biopanning and Rapid Analysis of Selective Interactive Ligands). After three rounds of selection, two highly reactive clones to the papillary thyroid tumor cell line NPA were further evaluated, and their specific binding to tumor proteins was confirmed using phage-ELISA. The antibody-like peptide CaT12 was tumor-specific, which was further tested by immunohistochemistry against TMAs (tissue microarrays) comprised of 775 human benign and malignant tissues, including 232 thyroid nodular lesions: 15 normal thyroid tissues, 53 nodular goiters (NG), 54 follicular adenomas (FA); 69 papillary thyroid carcinomas (PTC); and 41 follicular carcinomas (FC). CaT12 was able to identify PTC among thyroid nodular lesions with 91.2% sensitivity and 85.1% specificity, despite its non-specificity for thyroid tissues. Additionally, the CaT12 peptide helped characterize follicular lesions distinguishing the follicular variant of PTC (FVPTC) from FA with 91.9% accuracy; FVPTC from NG with 83.1% accuracy; FVPTC from the classic PTC with 57.7% accuracy; and FVPTC from FC with 88.7% accuracy. In conclusion, our strategy to select differentially expressed ligands to thyroid tissue was highly effective and resulted in a useful antibody-like biomarker that recognizes malignancy among thyroid nodules and may help distinguish follicular patterned lesions.
    Cancer letters 02/2013; · 5.02 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Abstract Recombinant antibodies are increasingly being employed as therapeutic agents especially in combination with anti-cancer drugs. The single-chain antibody fragments are small antigen-binding proteins which provide the most commonly used antibody formats for diagnostic and therapeutic purposes. These antibody fragments have more rapid tumor penetration and clearance from the serum relative to full-length monoclonal antibodies. There are in vitro antibody-display technologies such as phage display, cell surface display, ribosome display and mRNA display that can be used to isolate high specificity and affinity single-chain antibodies against a wide variety of targets. We review these strategies for generation of stable and active antibody fragments in the present article.
    Immunopharmacology and Immunotoxicology 08/2014; · 1.36 Impact Factor