Screening of phage clones isolated from the α-helix & 3₁₀-helix libraries.

<p>A) Monoclonal scFv screening ELISA testing 96 clones in each library. Bacterial supernatants were added to 5 µg/ml recombinant αvβ6 immobilized onto ELISA plate and then probed with mouse anti-Myc antibody followed by anti-mouse-HRP. B) Clones with unique sequences were screened for binding cellular αvβ6 by flow cytometry. Figure shows examples where scFv was tested at 100 (red histogram), 10 (orange histogram) and 1 (green histogram) µg/ml. For clarity the relevant αvβ6-specific mouse monoclonal antibody 10D5 (grey) and the negative control IgG (black) histograms are also shown in each plot. C) Binding to cellular αvβ6 by D25 and D34 verified by flow cytometry at 100 (red histogram), 10 (orange histogram) and 1 (green histogram) µg/ml. Size-exclusion chromatography profile of purified of D25scFv and D34scFv showed a major peaks at 30 kDa.