The technology behind Fluidic Analytics instruments allows for the characterisation of physiologically relevant protein interaction behaviour in tag-free systems:
Emma V. Yates, Thomas Müller, Luke Rajah, Erwin J. De Genst, Paolo Arosio, Sara Linse, Michele Vendruscolo, Christopher M. Dobson, Tuomas P.J. Knowles. Latent Analysis of unmodified biomolecules and their complexes in solution with attomole detection sensitivity. Nature Chemistry 7(10):802-9, 2015.
The study of biomolecular interactions is central to an understanding of function, malfunction and therapeutic modulation of biological systems, yet often involves a compromise between sensitivity and accuracy. Many conventional analytical steps and the procedures required to facilitate sensitive detection, such as the incorporation of chemical labels, are prone to perturb the complexes under observation. Here we present a ‘latent’ analysis approach that uses chemical and microfluidic tools to reveal, through highly sensitive detection of a labelled system, the behaviour of the physiologically relevant unlabelled system. We implement this strategy in a native microfluidic diffusional sizing platform, allowing us to achieve detection sensitivity at the attomole level, determine the hydrodynamic radii of biomolecules that vary by over three orders of magnitude in molecular weight, and study heterogeneous mixtures. We illustrate these key advantages by characterizing a complex of an antibody domain in the solution phase and under physiologically relevant conditions.
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The technology behind Fluidic Analytics instruments allows for the analysis of size and interactions between proteins:
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. Knowles. Microfluidic diffusion analysis of the sizes and interactions of proteins under native solution conditions. ACS Nano 10 333-341, 2015.
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.
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The technology behind Fluidic Analytics instruments allows for the measurement of viscosity:
Paulo Arosio, Kevin Hu, Francesco A. Aprile, Thomas Muller, Tuomas P.J. Knowles. Microfluidic diffusion viscometer for rapid analysis of complex solutions. Anal. Chem 88 (7) 488-3493, 201.6
The viscosity of complex solutions is a physical property of central relevance for a large number of applications in material, biological, and biotechnological sciences. Here we demonstrate a microfluidic technology to measure the viscosity of solutions by following the advection and diffusion of tracer particles under steady-state flow. We validate our method with standard water-glycerol mixtures, and then we apply this microfluidic diffusion viscometer to measure the viscosity of protein solutions at high concentrations as well as of a crude cell lysate. Our approach exhibits a series of attractive features, including analysis time on the order of seconds and the consumption of a few μL of sample, as well as the possibility to readily integrate the microfluidic viscometer in other instrument platforms or modular microfluidic devices. These characteristics make microfluidic diffusion viscometry an attractive approach in automated processes in biotechnology and health-care sciences where fast measurements with limited amount of sample consumption are required.
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