title: A novel microfluidic mixer based on dual-hydrodynamic focusing for interrogating the kinetics of DNA-protein interaction.
abstract: Kinetic measurement of biomacromolecular interaction plays a significant role in revealing the underlying mechanisms of cellular activities. Due to the small diffusion coefficient of biomacromolecules, it is difficult to resolve the rapid kinetic process with traditional analytical methods such as stopped-flow or laminar mixers. Here, we demonstrated a unique continuous-flow laminar mixer based on microfluidic dual-hydrodynamic focusing to characterize the kinetics of DNA-protein interactions. The time window of this mixer for kinetics observation could cover from sub-milliseconds to seconds, which made it possible to capture the folding process with a wide dynamic range. Moreover, the sample consumption was remarkably reduced to <0.55 μL min⁻¹, over 1000-fold saving in comparison to those reported previously. We further interrogated the interaction kinetics of G-quadruplex and the single-stranded DNA binding protein, indicating that this novel micromixer would be a useful approach for analyzing the interaction kinetics of biomacromolecules.
authors.affiliation: Britton Chance Center for Biomedical Photonics at Wuhan National Laboratory for Optoelectronics-Hubei Bioinformatics & Molecular Imaging Key Laboratory, Systems Biology Theme, Department of Biomedical Engineering, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Li
authors.forename: Fei
authors.initials: F
authors.lastname: Xu
authors.forename: Chao
authors.initials: C
authors.lastname: Liu
authors.forename: Youzhi
authors.initials: Y
authors.lastname: Xu
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Feng
authors.forename: Bi-Feng
authors.initials: BF
authors.lastname: Liu
mesh_terms: D004247:DNA / Q000032:analysis / Q000493:pharmacokinetics*; D004268:DNA-Binding Proteins / Q000032:analysis / Q000493:pharmacokinetics*; D004780:Environment, Controlled; D057446:Hydrodynamics; D008163:Luminescent Measurements / Q000379:methods; D044085:Microfluidics / Q000379:methods*; D011485:Protein Binding / Q000502:physiology
chemical_list: D004268:DNA-Binding Proteins; D004247:DNA