Figure 2
Summary of the anti-NP antibody discovery results. (A) The pie chart in the left-hand side shows the distribution of a total of 753 positive (ELISA OD450nm > 0.5 binding to Protein A/L and to corresponding target NP) anti-NP monoclonal scFvs from the Step 3 of the antibody discovery procedure in Fig. 1B. The pie chart in the right-hand side shows the distribution of the 25 scFvs reformatted to IgG1 in the Step 4 of the antibody discovery procedure in Fig. 1B. These 25 IgGs were selected to form a panel of antibodies to represent the 753 scFvs according to the NP binding specificity groups as shown in (B) of this figure. The sequences of these 25 scFvs are listed in Supplementary Table S1. (B) The heat map shows the ELISA OD (450 nm) of the 753 anti-NP scFvs (y-axis) from the phage display selection cycles plotted against the 6 representative NPs (x-axis). The representative set of 25 scFvs are marked next to the heat map. The heat map and the clustering of the scFvs as shown by the dendrograms in the x-axis and y-axis were generated with the gplots package of the R software7,35. The experimental procedure for the ELISA measurement of the scFv-NP interactions is described in Methods.
