The Liquid Extraction Surface Analysis (LESA) capability of the TriVersa NanoMate enables simple, direct nanoESI mass spectrometric analysis from a variety of surfaces.
The new LESAPLUS allows for automated LESA experiments plus additional nano-LC separation through the ChipSoftX operating software with Developers Kit. This enhancement is ideal for direct tissue analysis.
ChipSoftX is an entirely new operating software for the TriVersa NanoMate automated nanoelectrospray source. Besides improvement in program compatibility with Windows and integration of existing software features, it also provides access to the new Developers Kit – a platform for customized method development with direct access to robot controls allowing entirely novel analysis workflows such as LESAPLUS.
Native mass spectrometry seeks to probe noncovalent protein interactions in terms of protein quaternary structure, protein–protein and protein–ligand complexes. The ultimate goal is to link the understanding of protein interactions to the protein environment by visualizing the spatial distribution of noncovalent protein interactions within tissue. Previously, we have shown that noncovalently bound protein complexes can be directly probed via liquid extraction surface analysis from dried blood spot samples, where hemoglobin is highly abundant. Here, we show that the intact hemoglobin complex can be sampled directly from thin tissue sections of mouse liver and correlated to a visible vascular feature, paving the way for native mass spectrometry imaging.
R.L. Griffiths and H.J. Cooper Anal. Chem., 2016, 88 (1), pp 606–609
Analyst, 2015, Accepted Manuscript DOI: 10.1039/C5AN00933B
Liquid extraction surface analysis (LESA) is a surface sampling technique that allows electrospray mass spectrometry analysis of a wide range of analytes directly from biological substrates. Here, we present LESA mass spectrometry coupled with high field asymmetric waveform ion mobility spectrometry (FAIMS) for the analysis of dried blood spots on filter paper. Incorporation of FAIMS in the workflow enables gas-phase separation of lipid and protein molecular classes, enabling analysis of both haemoglobin and a range of lipid (phosphatidylcholine or phosphatidylethanolamine, and sphingomyelin species) from a single extraction sample. The work has implications for multiplexed clinical assays of multiple analytes.
Young, LM, Saunders, JC, Mahood, RA, Revill, CH, Foster, RJ, Tu, LH, Raleigh, DP, Radford, SE, Ashcroft, AE
Nature Chemistry7,73–81doi:10.1038/nchem.2129
The search for therapeutic agents that bind specifically to precursor protein conformations and inhibit amyloid assembly is an important challenge. Identifying such inhibitors is difficult because many protein precursors of aggregation are partially folded or intrinsically disordered, which rules out structure-based design. Furthermore, inhibitors can act by a variety of mechanisms, including specific or nonspecific binding, as well as colloidal inhibition. Here we report a high-throughput method based on ion mobility spectrometry–mass spectrometry (IMS–MS) that is capable of rapidly detecting small molecules that bind to amyloid precursors, identifying the interacting protein species and defining the mode of inhibition. Using this method we have classified a variety of small molecules that are potential inhibitors of human islet amyloid polypeptide (hIAPP) aggregation or amyloid-beta 1-40 aggregation as specific, nonspecific, colloidal or non-interacting. We also demonstrate the ability of IMS–MS to screen for inhibitory small molecules in a 96-well plate format and use this to discover a new inhibitor of hIAPP amyloid assembly.
Randall EC, Bunch J, Cooper HJ; Anal Chem. 2014 Nov 4;86(21):10504-10. doi: 10.1021/ac503349d. Epub 2014 Oct 23
Top-down identification of proteins by liquid extraction surface analysis (LESA) mass spectrometry has previously been reported for tissue sections and dried blood spot samples. Here, we present a modified “contact” LESA method for top-down analysis of proteins directly from living bacterial colonies grown in Petri dishes,without any sample pretreatment. It was possible to identify a number of proteins by use of collision-induced dissociation tandem mass spectrometry followed by searches of the data against an E. coli protein database. The proteins identified suggest that the method may provide insight into the bacterial response to environmental conditions. Moreover, the results show that the “contact” LESA approach results in a smaller sampling area than typical LESA, which may have implications for spatial profiling.
Martin NJ, Griffiths RL, Edwards RL, Cooper HJ. J Am Soc Mass Spectrom. 2015 May 20. [Epub ahead of print]
Liquid extraction surface analysis (LESA) mass spectrometry is a promising tool for the analysis of intact proteins from biological substrates. Here, we demonstrate native LESA mass spectrometry of noncovalent protein complexes of myoglobin and hemoglobin from a range of surfaces. Holomyoglobin, in which apomyoglobin is noncovalently bound to the prosthetic heme group, was observed following LESA mass spectrometry of myoglobin dried onto glass and polyvinylidene fluoride surfaces. Tetrameric hemoglobin [(αβ)2 4H] was observed following LESA mass spectrometry of hemoglobin dried onto glass and polyvinylidene fluoride (PVDF) surfaces, and from dried blood spots (DBS) on filter paper. Heme-bound dimers and monomers were also observed. The ‘contact’ LESA approach was particularly suitable for the analysis of hemoglobin tetramers from DBS.
Reinaldo Almeida, Zane Berzina, Eva C. Arnspang, Jan Baumgart, Johannes Vogt, Robert Nitsch, and Christer S. Ejsing
Here we describe a novel surface sampling technique termed pressurized liquid extraction surface analysis (PLESA), which in combination with a dedicated high resolution shotgun lipidomics routine enables both quantification and in-depth structural characterization of molecular lipid species extracted directly from tissue sections.
Development and applications of lipidomic technology that allow global analysis of cellular lipidomes with the ultimate goal of improving the understanding of the regulation of lipid metabolism.
I used the TriVersa NanoMate in Andrej Schevchenko’s laboratory at Max Planck Institute in Dresden. I learned that the instrument was far easier to use than nanoelectrospray needles. Additionally, the TriVersa NanoMate o ffers high throughput and is easy to teach others, especially students, how to use. You can rely on the TriVersa NanoMate to get the job done.
The TriVersa NanoMate is essential for any laboratory interested in lipidomics research for its ease of use, reliability and stable ion spray.
Lipid Molecular Timeline Profiling Reveals Diurnal Crosstalk between the Liver and Circulation
Sprenger et al. Cell Rep. 2021 34(5):108710
In-depth lipidomics and time-series analysis of fasting-feeding cycles and circadian rhythms provides insights into metabolic crosstalk between tissues.
Surface analysis techniques have gained significant attention over the last couple of years with LESA being an interesting alternative to techniques such as SIMS or MALDI imaging. This note demonstrates practical applications for LESA in the material sciences.