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. 2017 Oct 31;7:14455. doi: 10.1038/s41598-017-14823-w

Figure 3.

Figure 3

High throughput screening of anti-IAV H1N1 CA/09 neutralizing scFvs binding to H1N1 CA/09 HA trimer. (a) High throughput screening results (Step 3~4 in Fig. 1) for more than 5000 monoclonal scFvs with the protocol of Step 1a in Fig. 1 are summarized in this panel. The y-axis shows the ELISA signal of the monoclonal scFvs binding to H1N1 CA/09 HA trimer; the x-axis shows the relative viral activity based on the viral activity ratio measured in the microneutralization assay with IAV H1N1 CA/09 after and before addition of the scFv culture medium to the assay system – 100% relative viral active means no inhibition of the scFv to the IAV infection to the MDCK cell; 0% relative viral activity means complete inhibition of the infection. Positive control scFvs derived from F10 and FI6v3 IgGs show almost 0% relative viral activity in the assay and negative control scFv derived from AV1 IgG shows almost 100% relative viral activity. The dark blue data points labelled with antibody names are selected for IgG reformatting (Supplementary Tables S3~S4). The error bars associated with these data points were determined with at least three repeats of the microneutralization measurements. (b) The description is the same as in panel (a) for the high throughput screening results (Step 3~4 in Fig. 1) for more than 100 monoclonal scFvs with the protocol of Step 1b in Fig. 1 are summarized in this panel. Experimental details are described in Methods.