Fig. 3: NGS enrichment ranking selection of CD151 in situ selection of Ab clones. | Communications Biology

Fig. 3: NGS enrichment ranking selection of CD151 in situ selection of Ab clones.

From: CellectSeq: In silico discovery of antibodies targeting integral membrane proteins combining in situ selections and next-generation sequencing

Fig. 3

The abundance of each sequence (N:105,434) in Fab-phage pools selected for binding to HEK293T-CD151+ cells (positive counts, x-axis) is plotted versus the ratio of the abundance in pools selected for binding to HEK293T-CD151+ cells over pools selected for binding to HEK293T-CD151− (positive/negative, y-axis). Each circle represents one unique paratope (i.e., unique combination of CDRs L3, H1, H2, and H3). The dashed red lines define an upper-right quadrant that contains putative CD151 binding clones, defined arbitrarily as those occurring more than 200 times in the positive pool and being greater than fourfold enriched relative to the negative pool. The red circles represent the 100 Ab clones in the upper-right quadrant that were selected after the NGS analysis and predicted to bind to CD151. All selected clones are close homologs (>80% sequence identity) of the immunodominant Ab clone CD151-1. The red circle at the far right represents the immunodominant Ab clone CD151-1 that was manually sampled and validated as an anti-CD151 Ab by cellular phage ELISA (Fig. S5).

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