In vitro high throughput phage display selection of ovarian cancer avid phage clones for near-infrared optical imaging

• Published in: Combinatorial chemistry & high throughput screening Vol. 17; no. 10; p. 859
• Main Authors: Soendergaard, Mette, Newton-Northup, Jessica R, Deutscher, Susan L
• Format: Journal Article
• Language: English
• Published: United Arab Emirates 01.01.2014
• Subjects: Amino Acid Sequence; Animals; Bacteriophages - chemistry; Bacteriophages - metabolism; Cell Line; Female; HEK293 Cells; High-Throughput Screening Assays; Humans; Mice, Nude; Microscopy, Fluorescence; Molecular Sequence Data; Optical Imaging; Ovarian Neoplasms - diagnosis; Ovary - pathology; Peptide Library; Peptides - chemistry; Peptides - metabolism
• ISSN: 1875-5402
• DOI: 10.2174/1386207317666141031152828

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.