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Analysis of “difficult” proteins by microfluidic-based hydrogen/deuterium exchange

Project description

Innovative chip technology to improve mass-spectrometry of protein systems

Many important proteins cannot be studied by traditional structural analysis methods as they are unstable, too large or present in limited quantities in their native state. A promising approach is to use mass spectrometry to measure the hydrogen/deuterium exchange of proteins (HDX-MS). The EU-funded HDXchip project aims to develop innovative microchips to revolutionise the HDX-MS method to enable the analysis of protein systems of very high complexity. This technology will enable improvements in HDX-MS applications, such as analysis of the conformations and interactions of large protein complexes and analysis of proteins inside their lipid membranes. The project's success may dramatically improve understanding of the function of such proteins and their potential for targeting by drugs.

Objective

Many proteins important in biology or disease are not amenable to traditional methods for structural analysis as they cannot be isolated in sufficient amounts nor studied directly in their native environment. Alternative techniques are needed to understand the conformation and interactions of such difficult proteins and further support the on-going shift to protein-based ‘biopharmaceuticals’ in the pharmaceutical industry. One promising technique to meet this challenge is the use of mass spectrometry to measure the hydrogen/deuterium exchange of proteins (HDX-MS). However, HDX-MS suffers from significant limitations to realize its full potential and applicability.
HDXchip aims to develop innovative microchips to revolutionize the HDX-MS method and enable analysis of difficult protein systems of unprecedented complexity. HDXchip will create “HDX-MS 2.0” technology that integrates on-chip sample treatment and separation to overcome major limitations of the conventional method, including virtually eliminating unwanted deuterium loss (back-exchange). The key to fabrication of HDXchips lie in the fact that all elements will be made of thiol-ene polymers. These polymers are not only used to make the chip itself in a cheap and simple manner, but also to fabricate high surface area functionalities for fast sample treatment (incl. enzymatic reactions) and separation at unprecedented efficiency.
HDXchip will enable step-change improvements in popular HDX-MS applications, like analysis of the conformation and interactions of purified proteins and also open up for exciting and hitherto unfeasible applications such as analysis of proteins inside whole cells and proteins in their native lipid membranes. Hence, HDXchip could enable breakthrough insights into the conformation of difficult protein systems both in isolation and in their native environment, and dramatically improve our understanding of the function of such proteins and their potential to be targeted by drugs.

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Topic(s)

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ERC-COG - Consolidator Grant

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Call for proposal

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(opens in new window) ERC-2020-COG

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Host institution

KOBENHAVNS UNIVERSITET
Net EU contribution

Net EU financial contribution. The sum of money that the participant receives, deducted by the EU contribution to its linked third party. It considers the distribution of the EU financial contribution between direct beneficiaries of the project and other types of participants, like third-party participants.

€ 1 999 752,00
Address
NORREGADE 10
1165 KOBENHAVN
Denmark

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Region
Danmark Hovedstaden Byen København
Activity type
Higher or Secondary Education Establishments
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Total cost

The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.

€ 1 999 752,00

Beneficiaries (1)

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