TriVersa NanoMate® LESA® with ESI Chip® Technology

The TriVersa NanoMate® LESA® is the latest in chip-based electrospray ionization technology from Advion Interchim Scientific®. It combines the benefits of liquid chromatography, mass spectrometry, chip-based infusion, fraction collection and direct surface analysis into one integrated ion source platform. It allows scientists to obtain more information from complex samples than LC/MS alone.

CHIPSOFT 10.0 WITH DEVELOPERS KIT: Customized method development for the TriVersa Nanomate

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.


Direct Tissue Profiling of Protein Complexes: Toward Native Mass Spectrometry Imaging

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

Screening and classifying small-molecule inhibitors of amyloid formation using ion mobility spectrometry–mass spectrometry

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.

Native Liquid Extraction Surface Analysis Mass Spectrometry: Analysis of Noncovalent Protein Complexes Directly from Dried Substrates

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.

Institute for Research in Biomedicine (IRB Barcelona), Spain

What is the focus of your lab’s research?

Our goal is to provide the research community at IRB Barcelona and their co-workers with state-of-the-art tools and methodologies for the MS analysis of a broad range of biological species, from large proteins and DNA to small molecules. The final purpose is to get insight into these molecules’ identity, structure, interaction with other molecules and biological function in order to help in drug design, protein mechanism elucidation and in the search for biomarkers. We have implemented methods specialized in top-down proteomics and we are pioneers in this MS strategy in Spain.

As a core facility, we are responsible for working with different biologic molecules, and we are required to change methods constantly and efficiently.

How does the TriVersa NanoMate® align with your research goals?

Originally, we purchased the TriVersa NanoMate® for its chip-based direct infusion mode for noncovalent interaction analysis, but we learned quickly that it could be applied to other areas of our research. Prior to using the TriVersa NanoMate®, the steps involved in collecting fractions were painful and time-consuming. With the TriVersa NanoMate®, we can run LC/fraction collection or infusion without changing the setup and wasting time with stabilization. We do not experience the problems typical with traditional nanoelectrospray sources.

One aspect of the TriVersa NanoMate® that impressed us was the ability to analyze complicated top-down samples with the LC compatibility. It is not possible to analyze these samples on an LC time scale, and the fraction collection capability allowed us to analyze in a way that was not possible previously.

To whom would you recommend the TriVersa NanoMate® for their research?

We use the TriVersa NanoMate® for everything; noncovalent interactions, top-down, middle-down, bottom-up, basic infusion, LC coupled to fraction collection. The instrument is useful in all of its different set-ups, especially without having to change sources and waiting for a stable spray.

The reliability of the system is one of the greatest benefits especially for people who have to change frequently between applications. We have found the spray sensing feature to be very valuable because we know our precious samples will not be lost.

Do you have any publications or presentations using the TriVersa NanoMate®?

Publication Highlight

Characterization of Human Sperm Protamine Proteoforms Through a Combination of Top-Down and Bottom-Up Mass Spectrometry Approaches
Soler-Ventura et al. J Proteome Res, 2020, 19(1), 221-237. DOI: 10.1021/acs.jproteome.9b00499
Identified the sperm protamine proteoforms profile, including their post-translational modifications, in normozoospermic individuals using a top-down MS approach and a proteinase-K-digestion-based bottom-up MS approach.

Other Publications
  • Arauz-Garofalo et al. Protamine characterization by top-down proteomics: Boosting proteoform identification with DBSCAN. Proteoms, 2021. DOI: 10.3390/proteomes9020021
  • Yero et al. The Pseudomonas aeruginosa substrate-binding protein Ttg2D functions as a general glycerophospholipid transporter across the periplasm. Comm Bio, 2021. DOI: 10.1038/s42003-021-01968-8
  • Molnar et al. The histone code reader PHD finger protein 7 controls sex-linked disparities in gene expression and malignancy in Drosophila. Sci Adv, 2019. DOI: 10.1126/sciadv.aaw7965
  • Nadal et al. Structure of the homodimeric androgen receptor ligand-binding domain. Nat Commun, 2017. DOI: 10.1038/ncomms14388
  • Testoni et al. Lack of glycogenin causes glycogen accumulation and muscle function impairment. Cell Metabolism, 2017. DOI: 10.1016/j.cmet.2017.06.008
  • Izquierdo-Serra et al. Optical control of endogenous receptors and cellular excitability using targeted covalent photoswitches. Nat Commun, 2016. DOI: 10.1038/ncomms12221
  • Pujol-Pina et al. SDS-PAGE analysis of Aβ oligomers is disserving research into Alzheimer’s disease: Appealing for ESI-IM-MS. Sci Rep, 2015. DOI: 10.1038/srep14809
  • Saez et al. Influence of PPh3 moiety in the anticancer activity of new organometallic ruthenium complexes. J Inorg Biochem, 2014. DOI: j.jinorgbio.2014.03.002
  • Borg et al. Spectral counting assessment of protein dynamic range in cerebrospinal fluid following depletion with plasma-designed immunoaffinity columns. Clin Proteomics, 2011. DOI: 10.1186/1559-0275-8-6