title: Microfluidic Diffusion Analysis of the Sizes and Interactions of Proteins under Native Solution Conditions.
abstract: Characterizing the sizes and interactions of macromolecules under native conditions is a challenging problem in many areas of molecular sciences, which fundamentally arises from the polydisperse nature of biomolecular mixtures. Here, we describe a microfluidic platform for diffusional sizing based on monitoring micron-scale mass transport simultaneously in space and time. We show that the global analysis of such combined space-time data enables the hydrodynamic radii of individual species within mixtures to be determined directly by deconvoluting average signals into the contributions from the individual species. We demonstrate that the ability to perform rapid noninvasive sizing allows this method to be used to characterize interactions between biomolecules under native conditions. We illustrate the potential of the technique by implementing a single-step quantitative immunoassay that operates on a time scale of seconds and detects specific interactions between biomolecules within complex mixtures.
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, U.K.
authors.forename: Paolo
authors.initials: P
authors.lastname: Arosio
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, U.K.|Fluidic Analytics Ltd. , Cambridge CB2 1EW, U.K.
authors.forename: Thomas
authors.initials: T
authors.lastname: Müller
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, U.K.
authors.forename: Luke
authors.initials: L
authors.lastname: Rajah
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, U.K.
authors.forename: Emma V
authors.initials: EV
authors.lastname: Yates
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, U.K.
authors.forename: Francesco A
authors.initials: FA
authors.lastname: Aprile
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, U.K.
authors.forename: Yingbo
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, U.K.
authors.forename: Samuel I A
authors.initials: SI
authors.lastname: Cohen
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, U.K.
authors.forename: Duncan A
authors.initials: DA
authors.lastname: White
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, U.K.
authors.forename: Therese W
authors.initials: TW
authors.lastname: Herling
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, U.K.
authors.forename: Erwin J
authors.initials: EJ
authors.lastname: De Genst
authors.affiliation: Department of Biochemistry and Structural Biology, Lund University , Box 124, SE221 00 Lund, Sweden.
authors.forename: Sara
authors.initials: S
authors.lastname: Linse
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, U.K.
authors.forename: Michele
authors.initials: M
authors.lastname: Vendruscolo
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, U.K.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Dobson
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, U.K.
authors.forename: Tuomas P J
authors.initials: TP
authors.lastname: Knowles
mesh_terms: D000818:Animals; D002417:Cattle; D004058:Diffusion; D005456:Fluorescent Dyes / Q000737:chemistry; D005934:Glucagon / Q000737:chemistry; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry; D006801:Humans; D057446:Hydrodynamics; D007118:Immunoassay; D046210:Microfluidic Analytical Techniques; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D008970:Molecular Weight; D012710:Serum Albumin, Bovine / Q000737:chemistry; D061905:Single-Domain Antibodies / Q000737:chemistry; D012996:Solutions; D013194:Staining and Labeling / Q000379:methods*; D014867:Water / Q000737:chemistry; D051844:alpha-Synuclein / Q000737:chemistry; D009764:o-Phthalaldehyde / Q000737:chemistry
chemical_list: D005456:Fluorescent Dyes; D018840:HSP70 Heat-Shock Proteins; D061905:Single-Domain Antibodies; D012996:Solutions; D051844:alpha-Synuclein; D014867:Water; D012710:Serum Albumin, Bovine; D009764:o-Phthalaldehyde; D005934:Glucagon
keywords: diffusion; immunoassay; interactions; polydispersity; proteins; size distribution