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. 2022 Sep 6;14(1):2115200. doi: 10.1080/19420862.2022.2115200

Figure 5.

Panel A: bar plot shows no binding to antigen for the parental clone whenever the antigen is pre-blocked with one of the selected clones, but binding remains when the antigen is pre-incubated with a control clone. Panel B: scatter plot with dissociation rates for each clone on the X-axis and association rates on the Y-axis shows that the association rates of the selected clones are similar to the parental clone while the dissociation rates are generally 2 orders of magnitude slower. Panel C: surface plasmon resonance sensorgrams for selected clones show very slow dissociation for all of them.

(a) Antigen binding to yeast displaying the parental scFv when competing with 24 affinity-matured clones (A01-A02, A04-A12, B01-B12, G05) and an unrelated scFv-Fc (control). Binding values are given by the median APC fluorescence of the yeast displaying population. There is no binding to the target by the parental clone whenever the antigen is pre-blocked with one of the selected clones, but binding is retained when the antigen is pre-incubated with a control clone. (b) Observed on-rates (ka) and off-rates (kd) for the parental and 24 affinity-matured clones (clone G05, which does not have the LCDR2 liabilities is highlighted in Orange). Reported values are the mean of at least 4 measurements. Isoaffinity curves (KD) are shown as dashed diagonal lines. (c) SPR sensorgrams for 24 affinity-matured clones. The name of the clones and calculated KD are shown in each plot.