High-Throughput Analysis of Protein–Protein Interactions in Picoliter-Volume Droplets Using Fluorescence Polarization

Droplet-based microfluidic systems have emerged as a powerful platform for performing high-throughput biological experimentation. In addition, fluorescence polarization has been shown to be effective in reporting a diversity of bimolecular events such as protein-protein, DNA-protein, DNA-DNA, receptor-ligand, enzyme-substrate, and protein-drug interactions. Herein, we report the use of fluorescence polarization for high-throughput protein-protein interaction analysis in a droplet-based microfluidic system. To demonstrate the efficacy of the approach, we investigate the interaction between angiogenin (ANG) and antiangiogenin antibody (anti-ANG Ab) and demonstrate the efficient extraction of dissociation constants (K(D) = 10.4 ± 3.3 nM) within short time periods.