title: In vitro high throughput phage display selection of ovarian cancer avid phage clones for near-infrared optical imaging.
abstract: Ovarian cancer is among the leading causes of cancer deaths in women, and is the most fatal gynecological malignancy. Poor outcomes of the disease are a direct result of inadequate detection and diagnostic methods, which may be overcome by the development of novel efficacious screening modalities. However, the advancement of such technologies is often time-consuming and costly. To overcome this hurdle, our laboratory has established a time and cost effective method of selecting and identifying ovarian carcinoma avid bacteriophage (phage) clones using high throughput phage display technology. These phage clones were selected from a filamentous phage fusion vector (fUSE5) 15-amino acid peptide library against human ovarian carcinoma (SKOV-3) cells, and identified by DNA sequencing. Two phage clones, pM6 and pM9, were shown to exhibit high binding affinity and specificity for SKOV-3 cells using micropanning, cell binding and fluorescent microscopy studies. To validate that the binding was mediated by the phage-displayed peptides, biotinylated peptides (M6 and M9) were synthesized and the specificity for ovarian carcinoma cells was analyzed. These results showed that M6 and M9 bound to SKOV-3 cells in a dose-response manner and exhibited EC50 values of 22.9 ± 2.0 μM and 12.2 ± 2.1μM (mean ± STD), respectively. Based on this, phage clones pM6 and pM9 were labeled with the near-infrared fluorophore AF680, and examined for their pharmacokinetic properties and tumor imaging abilities in vivo. Both phage successfully targeted and imaged SKOV-3 tumors in xenografted nude mice, demonstrating the ability of this method to quickly and cost effectively develop novel ovarian carcinoma avid phage.
authors.forename: Mette
authors.initials: M
authors.lastname: Soendergaard
authors.forename: Jessica R
authors.initials: JR
authors.lastname: Newton-Northup
authors.affiliation: Harry S. Truman Memorial Veterans Hospital, Columbia, MO 65201, USA. deutschers@missouri.edu.
authors.forename: Susan L
authors.initials: SL
authors.lastname: Deutscher
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001435:Bacteriophages / Q000737:chemistry / Q000378:metabolism; D002460:Cell Line; D005260:Female; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D008819:Mice, Nude; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D061848:Optical Imaging; D010051:Ovarian Neoplasms / Q000175:diagnosis*; D010053:Ovary / Q000473:pathology*; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000378:metabolism
chemical_list: D019151:Peptide Library; D010455:Peptides