Single‐molecule mass spectrometry

Keifer, David Z.; Jarrold, Martin F.

doi:10.1002/mas.21495

Cited by 77 publications

(91 citation statements)

References 155 publications

(233 reference statements)

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“…In this work we used CDMS where the m / z and charge of each ion are measured simultaneously, yielding the mass of each ion directly. CDMS is one of several single‐molecule mass spectrometry methods that can be used to analyze large and heterogeneous objects …”

Section: Resultsmentioning

confidence: 99%

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Measurement of the accurate mass of a 50 MDa infectious virus

Keifer

Motwani

Teschke

et al. 2016

Rapid Comm Mass Spectrometry

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RATIONALE Bacteriophage P22 is believed to contain a total of 521 copies of 9 different proteins and a 41,724 base pair genome. Despite its enormous size and complexity, phage P22 can be electrosprayed, and it remains intact in ultra-high vacuum where its molar mass distribution has been measured. METHODS Phage P22 virions were generated by complementation in Salmonella enterica and purified. They were transferred into 100 mM ammonium acetate and then electrosprayed. The masses of individual virions were determined using charge detection mass spectrometry. RESULTS The stoichiometry of the protein components of phage P22 is sufficiently well-known that the theoretical molar mass can be determined to within a narrow range. The measured average molar mass of phage P22, 52,180±59 kDa, is consistent with the theoretical molar mass and supports the proposed stoichiometry of the components. The intrinsic width of the phage P22 mass distribution can be entirely accounted for by the distribution of DNA packaged by the headful mechanism. CONCLUSIONS At over 50 MDa, phage P22 is the largest object with a well-defined molar mass to be analyzed by mass spectrometry. The narrow measured mass distribution indicates that the virions survive the transition into the gas phase intact.

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Section: Resultsmentioning

confidence: 99%

“…CDMS is one of several single-molecule mass spectrometry methods that can be used to analyze large and heterogeneous objects. [49] CONCLUSIONS We have measured the molar mass distribution for infectious phage P22. Phage P22 contains a total of 521 copies of 9 different proteins along with dsDNA.…”

Section: Resultsmentioning

confidence: 99%

Measurement of the accurate mass of a 50 MDa infectious virus

Keifer

Motwani

Teschke

et al. 2016

Rapid Comm Mass Spectrometry

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“…The image charge waveform also yields the particle time-of-flight (TOF) and velocity through the pickup providing the mass-to-charge ratio for fixed energy particles [15]. This method has been used in accelerator experiments since at least 1960 [9], and with the increasing interest in studies of massive biomolecules, cells and nanoparticles it has seen more recent applications in mass spectrometry measurements [16][17][18][19][20][21][22][23][24], with the state of the art recently described by Keifer and Jarrold [25]. In our own laboratory, we make use of a variant of this approach, applying a charge-pickup electrode in an electrostatic fast-ion beam trap to monitor the ion density oscillating in the trap and carry out Fourier-transform (FT) mass spectrometry on ensembles of molecular ions [26,27].…”

Section: Introductionmentioning

confidence: 99%

The aerosol impact spectrometer: a versatile platform for studying the velocity dependence of nanoparticle-surface impact phenomena

2017

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A new apparatus designed to accelerate/decelerate and study the surface impact phenomena of charged aerosols and nanoparticles over a wide range of mass-tocharge (m/z) ratios and final velocities is described. A nanoparticle ion source coupled with a linear electrostatic trap configured as an image charge detection (ICD) mass spectrometer allows determination of the mass-to-charge ratio and the absolute charge and mass of single nanoparticles. A nine-stage linear accelerator/ decelerator is used to fix the final velocity of the nanoparticles, and in the results reported here the coefficient of restitution for polystyrene latex spheres (PSLs) impacting on silicon is measured using ICD techniques. To enable this apparatus to study a wide range of m/z, the data acquisition system uses a transient digitizer interfaced to a field-programmable gate array module that allows real time calculation of m/z and determination of the pulse sequence for the linear accelerator/decelerator. Electrospray ionization of a colloidal suspension of PSL spheres of 510 and 990 nm has been used to demonstrate acceleration and deceleration of charged nanoparticles and the resolution of the apparatus. Measurements of the coefficient of restitution for PSLs on silicon over the range 10-400 m/s are consistent with previous studies.

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“…An attractive alternative may come from measuring one particle (or ion) at a time, thereby avoiding the problematic convolution of signals that stem from insufficient resolving power [17][18][19][20][21][22][23][24][25][26][27] . When such single-particle detection approaches can be combined with an independent measure of the charge of ions, or when masses can be estimated using entirely different physical principles that circumvent the need to work with multiply charged ions, this opens up the door to bona fide single-particle mass spectrometry measurements.…”

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confidence: 99%

“…Several techniques for single particle mass spectrometry, capable of determining masses into the MDa range have emerged in recent years, most notably Nano Electro-Mechanical System mass spectrometry (NEMS-MS) and Charge Detection mass spectrometry (CD-MS) 21,22 . In NEMS-MS, single particles are deposited on a NEMS resonator, whose vibrational frequency is affected by the absorption of a single particle.…”

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Resolving heterogeneous high-mass macromolecular machineries by Orbitrap-based single particle charge detection mass spectrometry

Wörner

Snijder

Bennett

et al. 2019

Preprint

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Here we show that single particle charge-detection mass spectrometry (CD-MS) can be performed on a ubiquitous Orbitrap mass analyser and applied to the analysis of highmass (megadalton) heterogeneous biomolecular assemblies. We demonstrate that single particle high-mass ions can survive in the Orbitrap for seconds, whereby their measured signal amplitudes scale linearly with charge state over the entire m/z range. Orbitrap based single particle CD-MS can be used to resolve mixed ion populations, accurately predict charge states, and consequently also the mass of the ions. We successfully applied CD-MS to challenging natural and biotherapeutic protein assemblies, such as IgM oligomers, designed protein nano-cages, ribosome particles and intact, empty-and genome-loaded Adeno-associated virus particles. Single particle CD-MS combined with native MS on existing Orbitrap platforms will greatly expand its application, especially in the mass analysis of megadalton heterogeneous biomolecular assemblies.3 Main text:Over the past two decades, native mass spectrometry (MS) has developed into a powerful tool for the characterization of biological macromolecular complexes 1 . These complexes are analysed from solvents that preserve the noncovalent interactions, enabling mass analysis of intact macromolecular assemblies into the megadalton (MDa) range 2 . The exact mass of the intact macromolecular complex is then used to infer its composition, the subunit stoichiometry, and even the number and chemical nature of post-translational modifications and small molecule ligands bound to the complex 3-6 . Various mass analysers, including Quadrupole Time-of-Flight, Fourier-Transform Ion CyclotronResonance, and most recently Orbitrap™-based mass analysers, have all been adapted for native MS experiments 7-12 . With the successful modifications of the Orbitrap for use in native MS, an unprecedented mass resolving power in the high mass range could be achieved, opening the avenue for high resolution analysis on ever more heterogeneous protein assemblies like therapeutic and plasma glycoproteins, designed protein cages, membrane protein assemblies, intact viruses, and ribosomes 12,13 .Notably, masses are not measured directly in most MS approaches, but need to be inferred from the mass-to-charge (m/z) ratios of the detected ions. As first shown in the pioneering work of Mann and Fenn 14 , the charge states from a population of multiply charged ions generated by electrospray ionization (ESI) can be determined from the m/z values by matching consecutive peaks in the charge state distribution to calculate accurate masses. A general limitation in native MS studies then stems from the fact that the charge state, and thus also the mass, can only be accurately measured when multiple charge states of the same molecular species can be resolved and assigned in the m/z for the trimer and 56 for the tetramer precisely coincide at 10,649 m/z (see Fig. 4B).However, the single ion intensities of the IgG oligomers scale with charge state (see Fig. 4C...

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Single‐molecule mass spectrometry

Cited by 77 publications

References 155 publications

Measurement of the accurate mass of a 50 MDa infectious virus

Measurement of the accurate mass of a 50 MDa infectious virus

The aerosol impact spectrometer: a versatile platform for studying the velocity dependence of nanoparticle-surface impact phenomena

Resolving heterogeneous high-mass macromolecular machineries by Orbitrap-based single particle charge detection mass spectrometry

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