ASMS 2020 Reboot

Friday, July 31, 2020

ASMS 2020 was one of the first events in the laboratory meeting segment to convert from face-2-face to a “virtual” format due to the SARS-CoV-2 pandemic. The live part of the meeting expanded to June 1 to 12, 2020. User’s sessions and off-program events started in late May and extended into mid-July. American Society for Mass Spectrometry (ASMS) staff rallied and developed a new plan which they called “ASMS Reboot 2020”, which utilized online web communication such as Zoom and Webinars for communication. The staff of the ASMS Reboot deserves special credit for making the very best of a bad situation.

My analysis: Reboot’s format is adequate for essential communication and almost a direct replacement for lectures, especially for those with large attendance such as keynotes. However, the virtual format is bland. Lecturers use pointers and cursors to guide the audience. Unfortunately, these audience aids are not widely used in web-based presentations. Busy slides can be hard to follow. Plus, side talks with the investigators and colleagues are missing. Another factor, it is difficult to block out interruptions in an office, especially a home office, which increases frustration with the format. The virtual format much less expensive and less risky to human health. However, it is sort of like comparing being in the audience of a travel slide show to the thrill of being on-site atop Mt. Fuji or visiting the Coliseum in Rome, or in the audience at a concert.

Exhibition for Reboot 2020

Traditionally, the focus of ASMS annual meetings is new mass spectrometry products and enabled applications. This review covers new products and enabled applications. The exhibition for Reboot 2020 was scheduled for only four hours on Friday, June 5. Several vendors reported product introductions were rescheduled for other meetings.

Aspect Analytics Highlights imaging MS

Imaging with mass spectrometry is revealing details of cellular structure that needs to be correlated with results from other technology including atomic force and rapidly expanding optical techniques. Aspect Analytics NV, (Genk, Belgium) exhibited their software to apply their bioinformatics, artificial intelligence and machine learning to mass spec imaging. Non-rigid registration facilitates comparisons between many tissues and specimens. Aspect’s workflow organizes the MS data tasks to support all stakeholders. Yet, the software is easy to use and supported so that no IT expertise is required of the users. Aspect sponsored a webinar about their metabolite explorer. The package was developed with the cooperation of Boehringer Ingelheim. Aspect is seeking other collaboration opportunities.

Biocrates Life Sciences AG Studies Organ Function and Disease Stratification

Biocrates Life Sciences AG, (Innsbruck, Austria) focused on metabolomics for stratification of infectious diseases including SARS-CoV-2. Severe infections affect other organs beyond the initial target. With multi-organ response, metabolomics produces biomarkers that can differentiate between different infections, which is useful in guiding therapy. For example, sphingolipids are associated with poor outcomes in lower respiratory infections. Tryptophan metabolites are helpful in guiding pneumonia therapy. A bile acid profile can guide fecal transplant therapy of C. difficile infections.

ESI Source Solutions, LLC Focused on Nano-LC/MS/MS

ESI Source Solutions, LLC (Woburn, MA), focuses on capillary and nanoscale chromatography for Thermo Scientific Instruments. ESI also distributes the Heat Wave Nanospray sources developed by CorSolutions, LLC (Ithaca, NY). The heat Wave facilitates nanospray separations at elevated temperatures. ABIRD stands for Active Background Ion Reduction Device for ESI MS and MS/MS separations. The ABIRD manages the Microenvironment of the nanospray, which improves the signal/noise ration significantly.

Biognosys Adds Features to Spectronaut Software

Biognosys (Schlieren, Switzerland) introduced the SpectroNaut ™14, which adds several new features to their proteomics software. Added features include Data Independent Analysis (DIA) that avoids the need to curate large data-dependent libraries that require sample fractionation. Deep learning with directDIA™ enhancements improved protein count by 25% compared to the prior release (SpectroNaut 13). It also simplifies the workflow.

Spectronaut 14 also uses ion mobility acquisition from Bruker’s (Billerica, MA) timsTOF Pro™, Thermo Fisher Scientific’s (Waltham, MA) FAIMS Pro ™, and Waters (Milford, MA) Synapt G-2-Si HDMSE. This facilitates label-free quantitation of mixtures. These advances were supported by nine posters and several lectures. For example, Prof. Brigit Schilling of The Buck Institute for Research on Aging (Novato, CA)) lectured on Data-Independent Acquisition Strategies to Gain Deep Coverage. Her lecture focused on biomarkers of cellular senescence using Data Independent Acquisition. Cellular senescence is a common change in humans characterized by irreversible growth arrest, resistance to apoptosis, and multi-faceted secretory phenotype. Upon transition to the senescent state, the cells secrete many new proteins. Using DIA, one can focus on sets of particular sets of precursor ions. She first had to show that senescence biomarkers are released into plasma. She found an overlap of 40 proteins of the core senescence-associated secretory phenotype (SASP) with a database of 220 significant biomarkers compiled by other groups. The study next focused on exosomes, which are the vector for intercellular communication. Plasma is a problematic matrix with a high background of peptides, IgG, hormones, etc. which were removed by steric exclusion chromatography. LC-MS further characterized the exosomes with a Thermo Extractive 480 MS using DIA and a pooled sample by DDA, which provided correlations and network analysis. This produced a list of proteins that change with aging. With the data, one can ask and evaluate many questions about the proteomics of human aging.

Evosep for Clinical LC-MS of Infections

ASMS 2020 was held on the web due to the SARS-CoV-2 pandemic. During the first six months of the epidemic, scientists and the public were wondering about the mechanism of the infection and disease progression. EvoSep (Odense C, DK) is a firm that developed a high throughput HPLC using short capillary columns packed with 1.5 um beads with C-18 surface chemistry. The companies vision is, “Robust and fast workflows are indispensable for successfully implementing large-cohort clinical plasma proteomics studies.” To meet this need, EVOSEP designed a mobile phase handling system that elutes the plasma sample from the disposable tip and elutes it into the HPLC. The presets are optimized for reproducibility with run-to-run % CV of 0.9sec, column to column % CV of 4.1 sec. and instrument to instrument % CV of 4.8 sec. This is sufficient for interlaboratory comparison and method transfer. The user simply selects the SPD (Samples Per Day) the range of 30 to 300. The EVOSEP One takes it from there. The 30 SPD provides the highest sequence coverage. The 300 SPD setting typically gives 60% coverage of a proteolytic digest sample.

Thermo Fisher Scientific introduces Exploris Series of Orbitraps

Thermo Fisher Scientific is challenging the dominance of triple quadrupole mass spectrometers for quantitative analysis with the introduction of the Exploris series of Orbitrap mass spectrometers, Table 1. Each Exploris is a compact benchtop instrument that is usually coupled to an HPLC for LC-MS. The Exploris 120 seems adequate for dedicated or targeted methods. The 480 is much faster and has significantly lower detection sensitivity. It also is compatible with more optional accessories such as ion sources.

Table 1: Comparison of selected specifications for Orbitrap Exploris Mass Spectrometers from Thermo Fisher Scientific introduced at ASMS 2020 Reboot.

Exploris Model	120	240	480
Item
Resolution @ m/z 200	120,000	240,000	480,000
Mass range m/z	40-3,000	40-6000	40-6000
Scan speed	Up to 22 Hz	Up to 22 Hz	Up to 40 Hz
Detection Sensitivity	200 fg	200 fg	50 fg

An accompanying poster showed label-free assay of a HELA cell digest (Pierce) of by LC-FAIMS (High Field Asymmetric waveform Ion Mobility Spectrometry). It is an atmospheric pressure ion mobility technique that separates gas-phase ions by their behavior in strong and weak electric fields. FAIMS is compatible with electrospray ionization. It complements LC and MS and LC/MS as an orthogonal separation technology, which is particularly useful in proteomic studies involving microheterogeneity and post-translational modifications. The authors found 5,379 proteins and 49,521 peptides in the digest using 60-minute gradient elution.

A comparison of results from four different instruments shows a 5.89% CV. The tight inter-instrument performance is attributed to a series of method development templates that include parameter presets optimized for particular applications such as gel spots, immunoprecipitation, TMT (Tandem Mass Tag) multiplexing and targeted PRM (Parallel Reaction Monitoring).

Thermo Fisher Scientific also described improvements in their paper spray ion source with the introduction of the VeriSpray™, which is ideal for dried blood spot analysis without chromatography. The sample on the paper is wetted and connected to a high voltage source, which causes the analytes to travel to the inlet of the MS: Simple, automatable, with a low environmental burden. Elisa assays were one featured example.

Orbitrap Eclipse Tribrid for Single-Cell Proteomics

Single-cell proteomics is designed to study cell heterogeneity. This requires the development of new workflows since one is manipulating tiny volumes. To meet this need, Thermo Fisher Scientific introduced the Orbitrap Eclipse Tribrid mass spectrometer with their FAIMS Pro™ interface and nanoPOTS sample processing platform that uses microfabricated multi-well plates which minimize sample loss due to dilution. A webinar described the detection and quantitation of more than 1000 proteins, including proteoforms from single mammalian cells.

An Update on Ion Mobility Spectrometry (IMS)

At ASMS 2019, Waters Corporation introduced a novel dedicated cyclic ion mobility instrument with a reflectron TOF MS for detector. The cyclic device allowed one to select the length of the drift tube in one-meter intervals. The longer the drift tube, the better the resolution. Previously, ion mobility had been used as an orthogonal stage in premium-priced MS instruments, but the drift tube length was about half a meter or less. Results were enticing and promising.

Preview of SLIM Ion Mobility MS from MOBILion Systems

Ion mobility seems to be emerging from a decades-long hibernation while other techniques caught up with the technology. Smith’s Detection did build a business testing aircraft passengers for volatile or semi-volatile organic compounds that could indicate potential passengers might have recently had contacted explosives.

Dr. Richard Smith of the Pacific Northwest Laboratory (Pullman, WA) described the basics of the SLIM technology at a CASSS organized meeting on Mass Spectrometry in 2014. Smith’s team showed that ions could be manipulated and stored for further study. They also showed that drift tubes could guide ions around corners without loss of resolution. This technology facilitated folded designs such as a 13-meter drift tube in a rectangle about the size of typing paper. Smith and colleagues have refined the ion manipulation technology, including combining it with a TOF mass spectrometer from Agilent.

In 2017, MOBILion Systems INC. was formed to commercialize this technology. Operations started in 2018. In 2019, in a 10,000 sqft facility in Chadds Ford, PA. Their first round of financing raised $15.4 million from investors, including Agilent Technologies (Santa Clara, CA). The firm showed shipping boxes for six prototype instruments destined for evaluators. They expect to formally introduce the instrument at ASMS 2021.

Waters Corporation

Waters announced improvements of their high-resolution mass spectrometers, including the XEVO G2-XS QTof, Synapt ™ XS, and SELECT Series Cyclic IMS Instruments. These should be most useful in life science research, including biopharma.

For the SELECT SERIES ™ Cyclic ™ IMS System, introduced at ASMS 2019, will soon be able to use Electron Capture Dissociation (ECD) to probe the higher-order structure of large proteins and peptides. ECD extends the IMS range to very large biopolymers. Waters Corporation introduced IMSⁿ technology, which involves fragmenting a biopolymer and then selecting a particular product and fragmenting it again, which is analogous to multiple reaction monitoring in mass spectrometry (MRM). Fragmentation with ECD is often orthogonal to collisional induced fragmentation (CID). ECD also often preserves the location of post-translational modifications. MRM is a beneficial technology in MS and should be similarly useful in Cyclic IMS systems. The ECD capability was developed in partnership with e-MSion, Inc. (Portland OR).

In addition to the ECD above, Waters Corporation introduced Desorption Electrospray Ionization DESI ™XS for imaging of the spatial distribution of molecules, including drugs, metabolites, and lipids in biological specimens. Intellectual property supporting DESI was licensed from Purdue Research Foundation and Prosolia, Inc. DESI is also marketed for the SYNAPT ™ XS and Xevo™ G2-XS QTof mass spectrometers. The new source is located in a partially sealed housing that isolates the sample from the lab environment and associated contamination issues. It also has a side camera, facilitating alignment of the source and sample.

Columns for LC-MS Waters Corporation Extends the Atlantis Line of HPLC Columns for LC/MS

The excellent detection sensitivity of LC-MS has revealed opportunities for improvement in HPLC Columns, particularly in the area of mixed-mode. The Atlantis C18AX is designed to improve the retention of neutral and ionic analytes. It operates over a pH range of 1 to 12 and avoids irreproducibility problems due to dewetting of the small pores.

The C18 AX column packing starts with a BEH spherical bead with nominal 1.7, 2.7, and 5 um, which covers the UPLC and HPLC size range. The porosity is 95 Å, which is smaller than most BEH particles but gives higher surface area and retention. Next, a quaternary amine is bonded to the BEH bead to provide anion exchange retention. Finally, a C-18 surface chemistry adds RPLC selectivity. Thus, one can use pH chemistry to augment the selectivity over a much wide pH range than mixed-mode chemistry bonded to silica.

Chromatography of anions is problematic due to the interaction of anions with Iron and titanium and other metals. This often shows up as poor mass balance or variable results for anions, especially chelating anions. Waters Corporation has developed a new metallic material for the column tube. It is not PEEK lined stainless steel. I expect that someone will do a laser ablation study to figure out what the new material is.

ChromaNik Improves pH Stability of Columns

ChromaNik Technologies, Inc. (Minato-ku, Osaka, Japan) is a developer and vendor of columns designed for LC-MS. The key features are low bleed, Wide pH operating range, and excellent reproducibility column to column. The firm also serves the needs of chromatographer for custom packed columns filled with stationary phases produced outside of Japan. The SunShell brand of core-shell stationary phase is end-capped (hexamethyltrisiloxane) core-shell silica phase that provides 50% more theoretical plates than columns packed with totally porous silica particles. Surface chemistries included C-18, C-8, PFP, phenyl, bonded to 2.0, 2.6, and 5.0-micron core-shell particles.

SunArmor is ChromaNik’s stationary phase for high pH operating range, covering pH 2 to 12. It shows excellent peak shape for acidic, basic, and chelating analytes. The Firms Sunniest brand uses a novel C-18 reagent and temperature treatment to anchor the C-18 to the particle with multipoint bonding chemistry and end-capped at high temperature. Other phases are available.

User’s Meetings

Traditionally, major vendors of MS have each organized and underwritten events for their users of their products at ASMS. The Reboot format changed the format with the vendors organizing multi-hour webinars usually with several speakers outside of the Reboot program. Some went on for several days.

Agilent Technologies, Inc. (Santa Clara, CA) presented a three day virtual MS smorgasbord on June 16-18, 2020 with descriptions of new systems using triple quadrupole technology and the rapid-fire instrument for ultra-high throughput.

LC-MS as an FDA Class 1 Medical Device

LC-MS instruments are complex systems with lots of idiosyncrasies. Thus, the FDA burdened them with research use only (RUO) classification. Yet, the results are potentially useful in diagnosing disease in humans. Plus, in other countries, LC-MS is routinely used in in-vitro diagnostics of humans. To bridge this chasm, Agilent introduced the K6460S Clinical Edition TQ LC/MS system designed to meet the FDA requirements for a Class 1 device for quantitative analysis in the clinical diagnostics market segment.

The LC starts with a Clinical Edition 1260 Infinity II HPLC coupled to a K6460 triple quadrupole Mass spectrometer. Agilent’s Mass Hunter Clinical Edition software package controls the instruments and facilitates report generation. Clinical Edition software facilitates simplification of workflows and eliminates the need for specialized training.

Agilent simplified the triple quadrupole product line by combing the best features of the 6470A and 6475 triple quadrupole platforms and added Agilent’s novel VacShield to get the Agilent 6470B Triple Quadrupole LC/MS. The VacShield reduces maintenance time by 92% since one does not need to break vacuum to clean the ion injector. This eliminates a six-hour workflow. The 6470B also incorporates the ion optics from 6495C Which improves detection sensitivity and mass range. The electronics have also been updated.

Agilent’s liquid handling stage for the RapidFire 400 has been upgraded to handle 1536 well plates in addition to legacy 96 or 384 well plates. The 400 no also has thermostatic control of the 64 plate sample hotel. Throughput can be as fast as 2 seconds for MS assays. If solid phase extraction (SPE/MS) is used, the cycle time can be as short as 8 sec/sample. The anticipated use of the RapidFire 400 is for very high throughput assays including screening and can process samples from 400 plates each with 1536 wells. Next, Agilent redesigned the layout to accommodate an Ultivo LC/TQ MS to save space and facilitate one keyboard control.

Bruker

Septic shock is one of the most frequent causes of death in the USA. This is especially true with SARS-CoV-2 where cytokine storms are blamed for many of the deaths. As part of the technical program, Bruker featured a webinar on their MALDI Biotype ® presented by William (Ike) Northern of CompuNet, Dayton OH). Usually, the Biotyper provides rapid identification of organisms in blood. However occasionally the results are ambiguous. Bruker has developed the MBT Sepsityper® solution that resolves the ambiguities for MRSA subtyping, Bacteroides fragilis cfia and KPC-producing Entrobacteriacae.

Bruker improves the detection sensitivity of MALDI by 10 to 100 X. Bruker introduces matrix-assisted laser desorption ionization (MALDI) with the launch of the world’s first commercial MALDI-2 post-ionization (PI) source. It a second laser irradiates the MALDI cloud to improve detection sensitivity by 10 to 100 X.

Sciex

Sciex (Framingham, MA) also hosted a three-day virtual event. The highlight was the introduction of the Echo MS system with a throughput of three samples per second in the MS only mode. Adding an SPE or chromatography front end to the Echo system slows it down to the pace for the separation.

As the Echo name indicates that LabCyte Echo acoustic liquid handling technology is used to route the sample solution into an orifice of a transfer line leading to the mass spectrometer. The reproducibility of the Echo MS is much better than when chromatography is required. The sample-to-sample carryover is very small. The Echo is probably enabled by Danaher’s acquisition of LabCyte in early 2019. Sciex is also in Danaher’s portfolio.

Shimadzu Scientific Instruments

Shimadzu Scientific Instruments (Colombia, MD) celebrated 50 years of evolution of mass spectrometry instruments dating back to the LKB-9000. This year they introduced the LCMS-8060 NX, a triple Quadrupole LC-MS that features world-class detection sensitivity and robustness. The new UFsweeper™ improves data quality by sweeping-out residues from the collision cell. This enables an MRM 9multiple reaction monitoring) speed of 555 channels/second. Thermal control of the ion source also improves robustness and reproducibility.

SSI responded quickly to SARS-CoV-2 with the introduction of the Covid-19 Drug Internal Standards kite that includes a certificate of analysis. They also introduced the 2019 Novel Corona Virus Detection Kit.

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