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Resolving protein mixtures using microfluidic diffusional sizing combined with synchrotron radiation circular dichroism
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Bortolini, Christian, Kartanas, Tadas, Copic, Davor, Morales, Itzel Condado, Zhang, Yuewen, Challa, Pavan K., Peter, Quentin A. E., Jávorfi, Tamas, Hussain, Rohanah, Dong, Mingdong, Siligardi, Giuliano, Knowles, Tuomas P. J. and Charmet, Jérôme (2019) Resolving protein mixtures using microfluidic diffusional sizing combined with synchrotron radiation circular dichroism. Lab on a Chip, 19 (1). pp. 50-58. doi:10.1039/C8LC00757H ISSN 1473-0197.
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WRAP-resolving-protein-mixtures-using-microfluidic-diffusional-sizing-combined-synchrotron-radiation-circular-dichroism-Charmet-2018.pdf - Accepted Version - Requires a PDF viewer. Download (7Mb) | Preview |
Official URL: https://doi.org/10.1039/C8LC00757H
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.
| Item Type: | Journal Article | |||||||||||||||||||||||||||||||||||||||
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| Subjects: | Q Science > QP Physiology | |||||||||||||||||||||||||||||||||||||||
| Divisions: | Faculty of Science, Engineering and Medicine > Science > Chemistry | |||||||||||||||||||||||||||||||||||||||
| Library of Congress Subject Headings (LCSH): | Proteins -- Spectra, Circular dichroism | |||||||||||||||||||||||||||||||||||||||
| Journal or Publication Title: | Lab on a Chip | |||||||||||||||||||||||||||||||||||||||
| Publisher: | Royal Society of Chemistry | |||||||||||||||||||||||||||||||||||||||
| ISSN: | 1473-0197 | |||||||||||||||||||||||||||||||||||||||
| Official Date: | January 2019 | |||||||||||||||||||||||||||||||||||||||
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| Volume: | 19 | |||||||||||||||||||||||||||||||||||||||
| Number: | 1 | |||||||||||||||||||||||||||||||||||||||
| Page Range: | pp. 50-58 | |||||||||||||||||||||||||||||||||||||||
| DOI: | 10.1039/C8LC00757H | |||||||||||||||||||||||||||||||||||||||
| Status: | Peer Reviewed | |||||||||||||||||||||||||||||||||||||||
| Publication Status: | Published | |||||||||||||||||||||||||||||||||||||||
| Access rights to Published version: | Restricted or Subscription Access | |||||||||||||||||||||||||||||||||||||||
| Date of first compliant deposit: | 15 November 2018 | |||||||||||||||||||||||||||||||||||||||
| Date of first compliant Open Access: | 22 November 2019 | |||||||||||||||||||||||||||||||||||||||
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