Figure 1.

ELISA analysis of phage binders to GHBP. (A) Binding of the H10 phage or its round-two frameshifting mutants to GHBP, anti-E-tag mAb, or nonfat milk-coated plates. Wells were coated with either 2% nonfat milk (M), 100 ng/well of GHBP (G), or 100 ng/well of anti-E-tag mAb (E). Phage were added at 10¹⁰/well and were detected by anti-M13 antibody–horseradish peroxidase conjugate. The clone name is indicated in the upper portions of the graphs. (B) Competition of H10 phage binding. Phage were added at 10¹⁰/well in the absence or presence of competitor. Competitors were GH, BSA, or the H10 synthetic peptide, added 1 h before phage. Phage were detected by anti-M13 antibody–horseradish peroxidase conjugate. Relative activity is defined as the ratio of ELISA A₄₀₅ values in the presence and the absence of the competitor. The A₄₀₅ for the control (no competitor) was 0.4 and is labeled M in the graph.