NGS of phage antibody outputs from whole-cell panning

We present a target-unbiased approach for antibody discovery that relies on generating mAbs against native target cell surfaces via phage display. This method combines a previously reported method for improved whole-cell phage display selections with next-generation sequencing analysis to efficiently identify mAbs with the desired target cell reactivity. This approach enabled the identification of three multiple myeloma cell surface antigens, and cognate monoclonal antibody probes. Rabbit phage antibody libraries were selected against human multiple myeloma cell lines, HEK293-ROR1-TetOn cells, and healthy donor-derived human PBMCs using the Fab-phage Biotinylation and Capture (FBC) method. Selected antibody outputs were subjected to NGS analysis for HCDR3 profiling.