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Resolving protein mixtures using microfluidic diffusional sizing combined with synchrotron radiation circular dichroism

DOI: 10.1039/C8LC00757H DOI Help

Authors: Christian Bortolini (University of Cambridge; Aarhus University) , Tadas Kartanas (University of Cambridge) , Davor Copic (University of Cambridge) , Itzel Condado Morales (University of Cambridge) , Yuewen Zhang (University of Cambridge) , Pavan K. Challa (University of Cambridge) , Quentin Peter (University of Cambridge) , Tamas Javorfi (Diamond Light Source) , Rohanah Hussain (Diamond Light Source) , Mingdong Dong (Aarhus University) , Giuliano Siligardi (Diamond Light Source) , Tuomas P. J. Knowles (University of Cambridge) , Jerome Charmet (University of Warwick)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Lab On A Chip , VOL 15

State: Published (Approved)
Published: November 2018
Diamond Proposal Number(s): 14750 , 15005 , 15592 , 16203

Abstract: Circular dichroism spectroscopy has become a powerful tool to characterise proteins and other biomolecules. For heterogeneous samples such as those present for interacting proteins, typically only average spectroscopic features can be resolved. Here we overcome this limitation by using free-flow microfluidic size separation in-line with synchrotron radiation circular dichroism to resolve the secondary structure of each component of a model protein mixture containing monomers and fibrils. To enable this objective, we have integrated far-UV compatible measurement chambers into PDMS-based microfluidic devices. Two architectures are proposed so as to accommodate for a wide range of concentrations. The approach, which can be used in combination with other bulk measurement techniques, paves the way to the study of complex mixtures such as the ones associated with protein misfolding and aggregation diseases including Alzheimer's and Parkinson's diseases.

Subject Areas: Biology and Bio-materials


Instruments: B23-Circular Dichroism