Fig. 2: Flow cytometry analysis of fluorescent labeled phage displayed Fabs (ph-Fabs) bound to ARS1620-V7 immobilized beads. | Communications Biology

Fig. 2: Flow cytometry analysis of fluorescent labeled phage displayed Fabs (ph-Fabs) bound to ARS1620-V7 immobilized beads.

From: Rare antibody phage isolation and discrimination (RAPID) biopanning enables identification of high-affinity antibodies against challenging targets

Fig. 2: Flow cytometry analysis of fluorescent labeled phage displayed Fabs (ph-Fabs) bound to ARS1620-V7 immobilized beads.

a ph-Fabs bound to ARS1620-V7-beads with varying incubation times. ph-P1A4 and ph-P2B2 were incubated for a total of 1 h and 3 h each. An increase in fluorescent intensity is observed beyond the 1-hour mark, indicating that the binding is not fully saturated even after 1 h of incubation. b Flow cytometry analysis of ph-Fabs of varying affinities bound to ARS1620-V7-beads. Fluorescent distributions of beads bound to ph-P2B2, ph-P1H6, ph-P1C1, and ph-P2F11 show MFI trends that agree with the ph-Fab affinities. c Flow cytometry analysis of increasing ph-P1A4 concentration shows an increase in normalized MFI with an observable shift from flow cytometry starting at 10 percent ph-P1A4 (d) Overlay of flow cytometry analysis of ph-P2B2, ph-P1H6, and a ph-P2B2, ph-P1H6 mix (90%/10%) bound to ARS1720-V7-beads. (e) Flow cytometry analysis of fluorescein isothiocyanate (FITC) and Alexa Fluor 647 (AF647) labeled mixture of ph-P2B2 and ph-P1H6. Pure ph-P2B2 labeled with both FITC and AF647 was used as a control. Histograms of the dot blot distributions are also shown.

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