Abstract
Ambient surface mass spectrometry is an emerging field which shows great promise for the analysis of biomolecules directly from their biological substrate. In this article, we describe ambient ionisation mass spectrometry techniques for the in situ analysis of intact proteins. As a broad approach, the analysis of intact proteins offers unique advantages for the determination of primary sequence variations and posttranslational modifications, as well as interrogation of tertiary and quaternary structure and protein‐protein/ligand interactions. In situ analysis of intact proteins offers the potential to couple these advantages with information relating to their biological environment, for example, their spatial distributions within healthy and diseased tissues. Here, we describe the techniques most commonly applied to in situ protein analysis (liquid extraction surface analysis, continuous flow liquid microjunction surface sampling, nano desorption electrospray ionisation, and desorption electrospray ionisation), their advantages, and limitations and describe their applications to date. We also discuss the incorporation of ion mobility spectrometry techniques (high field asymmetric waveform ion mobility spectrometry and travelling wave ion mobility spectrometry) into ambient workflows. Finally, future directions for the field are discussed.
Highlights
The interest in proteins is fuelled by their fundamental role in the biological processes occurring in living cells, both under normal conditions and in disease states
We discuss the incorporation of ion mobility spectrometry techniques into ambient workflows
We describe surface analysis techniques most commonly used for the mass spectrometric analysis of intact proteins, with emphasis on liquid extraction‐based techniques which, far, have proven most effective for intact protein analysis directly from biological substrates
Summary
The interest in proteins is fuelled by their fundamental role in the biological processes occurring in living cells, both under normal conditions and in disease states. KEYWORDS ambient mass spectrometry, surface sampling, intact proteins, ion mobility spectrometry, LESA, nanoDESI, DESI, Flowprobe
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