title: Protein Aggregate-Ligand Binding Assays Based on Microfluidic Diffusional Separation.
abstract: The measurement of molecular interactions with pathological protein aggregates, including amyloid fibrils, is of central importance in the context of the development of diagnostic and therapeutic strategies against protein misfolding disorders. Probing such interactions by conventional methods can, however, be challenging because of the supramolecular nature of protein aggregates, their heterogeneity, and their often dynamic nature. Here we demonstrate that direct measurement of diffusion on a microfluidic platform enables the determination of affinity and kinetics data for ligand binding to amyloid fibrils in solution. This method yields rapid binding information from only microlitres of sample, and is therefore a powerful technique for identifying and characterising molecular species with potential therapeutic or diagnostic application.
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.
authors.forename: Yingbo
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK. alexander.buell@uni-duesseldorf.de.|Institute of Physical Biology, University of Düsseldorf, Universitätsstrasse 1, 40225, Düsseldorf, Germany. alexander.buell@uni-duesseldorf.de.
authors.forename: Alexander K
authors.initials: AK
authors.lastname: Buell
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.|Fluidic Analytics Ltd, Unit 5 Chesterton Mill, French's Road, Cambridge, CB4 3NP, UK.
authors.forename: Thomas
authors.initials: T
authors.lastname: Müller
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.|AstraZeneca, Innovative Medicines, Discovery Sciences, Cambridge Science Park, Milton Road, Cambridge, CB4 0WG, UK.
authors.forename: Erwin
authors.initials: E
authors.lastname: De Genst
authors.affiliation: Department of Chemistry, Physical and Theoretical Chemistry Laboratory, University of Oxford, South Parks Road, Oxford, OX3 1QZ, UK.
authors.forename: Justin
authors.initials: J
authors.lastname: Benesch
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Dobson
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK. tpjk2@cam.ac.uk.
authors.forename: Tuomas P J
authors.initials: TP
authors.lastname: Knowles
mesh_terms: D000682:Amyloid / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D052160:Benzothiazoles; D001665:Binding Sites / Q000187:drug effects; D004058:Diffusion; D006801:Humans; D007700:Kinetics; D008024:Ligands; D046210:Microfluidic Analytical Techniques; D053758:Nanoparticles / Q000737:chemistry; D010316:Particle Size; D066329:Protein Aggregates / Q000187:drug effects; D057165:Proteostasis Deficiencies / Q000175:diagnosis* / Q000188:drug therapy; D013844:Thiazoles / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology
chemical_list: D000682:Amyloid; D052160:Benzothiazoles; D008024:Ligands; D066329:Protein Aggregates; D013844:Thiazoles; C009462:thioflavin T
keywords: aggregation; amyloid beta-peptides; fluorescent probes; microfluidic devices; protein-protein interactions