title: High-throughput analysis of protein-protein interactions in picoliter-volume droplets using fluorescence polarization.
abstract: 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.
authors.affiliation: Department of Biochemistry, Chungbuk National University, Cheongju, Republic of Korea.
authors.forename: Jae-Won
authors.initials: JW
authors.lastname: Choi
authors.forename: Dong-Ku
authors.initials: DK
authors.lastname: Kang
authors.forename: Hyun
authors.initials: H
authors.lastname: Park
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: deMello
authors.forename: Soo-Ik
authors.initials: SI
authors.lastname: Chang
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry; D005454:Fluorescence Polarization; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation; D011506:Proteins / Q000737:chemistry*; D012259:Ribonuclease, Pancreatic / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D011506:Proteins; C046366:angiogenin; D012259:Ribonuclease, Pancreatic