A: The mass spectrometry facility here at Boston College’s main focus is as a core lab, open to all users, both undergraduate and graduate students alike.
Our main research interests include the application of mass spectrometry as a routine tool for chemistry/chemical biology; exploring new research applications for mass spectrometry; and the use of open-air ionization methods for the analysis of pharmaceutical, biological, drug and chemical analysis.
Q: What was your previous work flow or challenges?
A: As a core lab we have a large number of samples being run and there is a constant need to change ionization sources.
Q: Why did you incorporate the expression® CMS into your laboratory?
A: The expression® has allowed us to take the workload of the other more expensive instruments by incorporating it as an open access instrument. With minimal training, the students are able to run their own samples, as not to slow down their research.
Due to the instrument size and speed at which it pumps down, we are able to move the instrument directly to the classroom for students to use.
A: The central activity of our group is to design and develop molecules to study and control biological systems. We are using organic and organometallic synthesis to construct new bioactive compounds, new agents for bioconjugation, and new probes for biology. Our focus is to apply chemistry to biology by identifying targets and then creating cancer treatment strategies and therapy options.
Q: WHAT PREVIOUS WORKFLOW CHALLENGES DID YOU EXPERIENCE?
A: For identification and analysis, we use NMR and HPLC regularly, but had no mass spectrometer in the laboratory, so we gave our samples to outside MS service for analysis. Therefore, we had to wait for the results.
Q: WHY DID YOU INCORPORATE THE EXPRESSION CMS INTO YOUR LABORATORY?
A: Advion’s CMS allows us to have a self-service option. One ingenieur is in-charge of the CMS, but everyone can use it including students after a short formation. And for 80% of our work it is perfect. We still use the MS facility for more demanding analysis, but this new workflow speeds up our process significantly.
Q: WHO WOULD YOU RECOMMEND TO PURCHASE THE EXPRESSION CMS?
A: I would recommend the CMS to any small or large organization like ours who wants fast answers for day-to-day analysis.
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
The purpose of this study was to evaluate the positive and negative electrospray (ESI) and atmospheric pressure chemical ionization (APCI) LC/MS ion current response from a representative synthetic mixture of standard pharmaceutical compounds. The selected drup compounds were prepared as an equimolar mixture and analyzed by an optimized gradient LC/MS procedure using a novel compact mass spectrometer. The total and extracted ion current chromatographic peaks were compared between ESI and APCI in both positive and negative ionization modes. The results from this study shed some light on the relative merits of APCI vs ESI as well as the importance of the optimal ionization polarity for the LC/MS determination of the selected acidic and basic small molecule drugs. We also note some differences in adduct formation between these two API techniques.
Developed in collaboration with Oak Ridge National Laboratory*, the liquid extraction surface analysis (LESA) capability of the TriVersa Nanomate® enables simple, direct ESI mass spectrometric analysis from a variety of surfaces.
A: Medicinal chemistry research in my lab is a collaborative exercise where we engage other scientists with complementary interests. Currently, we are engaged in the discovery of protein kinase inhibitors for anti-infective and anti-inflammatory applications and in the discovery of new agents useful for the potential treatment of neurodegenerative diseases. We are working with others at MSU as well as scientists in other universities and research institutes.
Q: Why did you incorporate the expression CMS into your laboratory?
A: As a medicinal chemist working in the industry, I wanted to have access to the same technology for my group’s research at MSU. However, working in the industry I had experts to run samples. In my lab at MSU, it is essential to have the same technology available in an easy to use the instrument. Not only do the expression CMS specifications meet our needs perfectly, but also it runs without a hitch and it is easily serviceable by a non-expert.
Q: Who would you recommend to purchase the expression CMS?
A: I would recommend the expression CMS to any non-expert chemist or laboratory who needs access to this level of technology. The Advion service and support groups make it accessible.
Presented by: John P. Shockcor, Director of Life Sciences Business Development, Waters Corp., Visiting Fellow, Dept. of Biochemistry, University of Cambridge, UK
Description: Profiling low level components in a complex mixture of small molecules can be a challenging task. Although it may be possible to detect many low level components in a complex mixture, characterization is often hindered because fragmentation of these low level components yields peaks below the limit of detection. This problem can be alleviated by using a TriVersa NanoMate assisted approach. In this webinar we will describe how a TriVersa NanoMate coupled to a SYNAPT G2 Hybrid QTof Ion-Mobility Mass Spectrometer can provide critical fragmentation information needed to characterize low level components in lipidomics, drug metabolism studies and natural product profiling. This approach is ideally suited to the use of time-aligned-parallel fragmentation (TAP) which are illustrated by a number of examples.
A: The development of macromolecular materials that combine synthetic polymerization techniques with the capacity of peptides and other biologically relevant molecules to participate in supramolecular interactions will lead to a variety of new responsive materials. When applied in vivo, a vital characteristic of these new materials will be their biodegradability, allowing the carrier to break down into benign metabolites. My lab focuses on the synthesis of biodegradable polymeric and self-assembling structures and their application in a broad variety of fields, including regenerative medicine, cancer therapeutics, drug delivery, and signaling gas delivery.
Q: What was your previous work flow or challenges?
A: We make and purify a lot of peptides, some of which are unstable under preparative HPLC purification conditions. By using rapid MS analysis to confirm the identity of an HPLC peak, we can quickly workup the product-containing fractions before they decompose. This was nearly impossible before we moved to the Advion system. Our peptides routinely have molecular weights in excess of 2000 g/mol, which we can identify by looking at multiply charged species. We use the expression® CMS for characterizing peptides several times during a difficult synthesis to optimize our synthesis and purification methods. My group has also found that the CMS system is extremely helpful in small molecule synthesis, where we use it to quickly verify that a product has formed in a reaction before proceeding to workup and isolation steps.
Q: Why did you incorporate the expression® CMS into your laboratory?
A: We incorporated the CMS system in our lab to speed up workflow and to enable us to synthesize peptides and peptide conjugates that are deliberately unstable. It has sped up small molecules and peptide synthesis as well as allowed us to do qualitative decomposition studies on our compounds.
Q: Who would you recommend to purchase the expression® CMS?
A: I think that nearly any lab that does significant small molecule and/or peptide synthesis would benefit from this instrument. There is no substitute for having a mass spectrometer a few steps away from your bench. Analyses require very little sample and are often as quick as a few minutes, making the CMS system a more efficient analytical technique than 1H NMR for rapid product confirmation.
The rapid identification of radiolabeled compounds would clearly be beneficial for applied clinical and preclinical tracers used for imaging with positron emission tomography (PET). The limited uptake of mass spectrometry in the imaging field has been due to several factors including concerns regarding adequate sensitivity, instrument footprint in the highly constrained space requirements for the majority of labs and high initial capital costs plus high ongoing maintenance costs.
The introduction of the expression Compact Mass Spectrometer (CMS) at the 2012 Spring ACS National Meeting addressed for the first time three of the concerns: space, capital cost and maintenance costs. Only the question of adequate sensitivity remained to be answered.
