nn5b04713_si_001.pdf (3.23 MB)
Microfluidic Diffusion Analysis of the Sizes and Interactions of Proteins under Native Solution Conditions
journal contribution
posted on 2015-12-23, 17:51 authored by Paolo Arosio, Thomas Müller, Luke Rajah, Emma V. Yates, Francesco
A. Aprile, Yingbo Zhang, Samuel I. A. Cohen, Duncan A. White, Therese W. Herling, Erwin J. De Genst, Sara Linse, Michele Vendruscolo, Christopher M. Dobson, Tuomas P. J. KnowlesCharacterizing
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.