Affinity ranking of antibodies using flow cytometry: Application in antibody phage display-based target discovery

We show that ranking of the antibodies based on FACS data correlates well with ranking based on KD values as measured by SPR and can therefore be used to discriminate between high- and low-affinity antibodies. Finally, we show that a low-affinity antibody may only detect high expression levels of a surface marker while failing to detect lower expression levels of this molecule, which may lead to a false interpretation of antibody specificity.