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

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

DOI

  • Christian Bortolini, Cambridge University
  • ,
  • Tadas Kartanas, Cambridge University
  • ,
  • Davor Copic, Cambridge University
  • ,
  • Itzel Condado Morales, Cambridge University
  • ,
  • Yuewen Zhang, Cambridge University
  • ,
  • Pavan K. Challa, Cambridge University
  • ,
  • Quentin Peter, Cambridge University
  • ,
  • Tamás Jávorfi, Diamond Light Source
  • ,
  • Rohanah Hussain, Diamond Light Source
  • ,
  • Mingdong Dong
  • Giuliano Siligardi, Diamond Light Source
  • ,
  • Tuomas P.J. Knowles, Cambridge University
  • ,
  • Jérôme Charmet, The University of Warwick

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.

Original languageEnglish
JournalLab on a Chip
Volume19
Issue1
Pages (from-to)50-58
Number of pages9
ISSN1473-0197
DOIs
Publication statusPublished - 7 Jan 2019

    Research areas

  • Animals, Cattle, Circular Dichroism/instrumentation, Diffusion, Equipment Design, Insulin/chemistry, Lab-On-A-Chip Devices, Particle Size, Protein Structure, Secondary, Proteins/analysis, Reproducibility of Results, Synchrotrons

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