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

Christian Bortolini; Tadas Kartanas; Davor Copic; Itzel Condado Morales; Yuewen Zhang; Pavan K. Challa; Quentin Peter; Tamás Jávorfi; Rohanah Hussain; Mingdong Dong; Giuliano Siligardi; Tuomas P.J. Knowles; Jérôme Charmet

doi:10.1039/c8lc00757h

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

Christian Bortolini, Tadas Kartanas, Davor Copic, Itzel Condado Morales, Yuewen Zhang, Pavan K. Challa, Quentin Peter, Tamás Jávorfi, Rohanah Hussain, Mingdong Dong, Giuliano Siligardi^*, Tuomas P.J. Knowles, Jérôme Charmet

^*Corresponding author for this work

Interdisciplinary Nanoscience Center

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

8 Citations (Scopus)

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.

Original language	English
Journal	Lab on a Chip
Volume	19
Issue	1
Pages (from-to)	50-58
Number of pages	9
ISSN	1473-0197
DOIs	https://doi.org/10.1039/c8lc00757h
Publication status	Published - 7 Jan 2019

Keywords

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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Bortolini, C., Kartanas, T., Copic, D., Condado Morales, I., Zhang, Y., Challa, P. K., Peter, Q., Jávorfi, T., Hussain, R., Dong, M., Siligardi, G., Knowles, T. P. J., & Charmet, J. (2019). Resolving protein mixtures using microfluidic diffusional sizing combined with synchrotron radiation circular dichroism. Lab on a Chip, 19(1), 50-58. https://doi.org/10.1039/c8lc00757h

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title = "Resolving protein mixtures using microfluidic diffusional sizing combined with synchrotron radiation circular dichroism",

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.",

keywords = "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",

author = "Christian Bortolini and Tadas Kartanas and Davor Copic and \{Condado Morales\}, Itzel and Yuewen Zhang and Challa, \{Pavan K.\} and Quentin Peter and Tam{\'a}s J{\'a}vorfi and Rohanah Hussain and Mingdong Dong and Giuliano Siligardi and Knowles, \{Tuomas P.J.\} and J{\'e}r{\^o}me Charmet",

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doi = "10.1039/c8lc00757h",

language = "English",

volume = "19",

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Resolving protein mixtures using microfluidic diffusional sizing combined with synchrotron radiation circular dichroism. / Bortolini, Christian; Kartanas, Tadas; Copic, Davor et al.
In: Lab on a Chip, Vol. 19, No. 1, 07.01.2019, p. 50-58.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - Resolving protein mixtures using microfluidic diffusional sizing combined with synchrotron radiation circular dichroism

AU - Bortolini, Christian

AU - Kartanas, Tadas

AU - Copic, Davor

AU - Condado Morales, Itzel

AU - Zhang, Yuewen

AU - Challa, Pavan K.

AU - Peter, Quentin

AU - Jávorfi, Tamás

AU - Hussain, Rohanah

AU - Dong, Mingdong

AU - Siligardi, Giuliano

AU - Knowles, Tuomas P.J.

AU - Charmet, Jérôme

PY - 2019/1/7

Y1 - 2019/1/7

N2 - 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.

AB - 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.

KW - Animals

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KW - Diffusion

KW - Equipment Design

KW - Insulin/chemistry

KW - Lab-On-A-Chip Devices

KW - Particle Size

KW - Protein Structure, Secondary

KW - Proteins/analysis

KW - Reproducibility of Results

KW - Synchrotrons

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DO - 10.1039/c8lc00757h

M3 - Journal article

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AN - SCOPUS:85058771775

SN - 1473-0197

VL - 19

SP - 50

EP - 58

JO - Lab on a Chip

JF - Lab on a Chip

IS - 1

ER -

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

Abstract

Keywords

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