Fig. 2
Generation and binding characterization of CLL-specific antibodies. a Schematic overview of the phage-display panning procedure. To enrich for CLL-specific antibodies, phages displaying scFv were mixed and incubated with a pool of CLL cells from multiple patients and a pool of PBMC from multiple healthy donors (with or without prior depletion of B-cells). Target (CLL) and non-target (PBMC) cells were then separated and phages binding CLL cells were eluted. b Screening in flow cytometry of 1152 scFvs’ binding to CLL cells from one patient versus binding to B-cell depleted PBMC from one healthy donor. c Heatmap showing the flow cytometry binding pattern of 392 unique scFvs to CLL cells from 5 patients, B-cells, and CD19-negative cells (B-cell depleted PBMC) from 2 healthy donors, B-cell lines (Raji and RPMI8226) and other cell lines (DU145, Lovo, MCF-7, and HS-5). Based on binding profile a hierarchal clustering of antibodies was made using QlucoreTM Omics Explorer. Clones were color-coded based on relative signal intensities within each cell type where red represents the strongest binding to a particular cell type, green the weakest binding and black in-between
