Charge detection mass spectrometry for the analysis of viruses and virus-like particles

Miller, Lohra M.; Jarrold, Martin F.

doi:10.1042/ebc20220101

Cited by 15 publications

(8 citation statements)

References 64 publications

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“…CD-MS extends the upper mass limit of MS by simultaneously measuring the m/z and z of individual ions to allow direct calculation of mass. CD-MS has been used to measure a number of viruses, 49 with recent utility in the analysis of rAAVs. 34 , 36 , 37 , 38 Prior to CD-MS analysis, rAAV samples were buffer exchanged into 200 mM ammonium acetate (Honeywell, 631-61-8) solution using Micro Bio-Spin columns (Bio-Rad, 7326221) in duplicate.…”

Section: Methodsmentioning

confidence: 99%

Partial genome content within rAAVs impacts performance in a cell assay-dependent manner

Troxell,

Jaslow,

Tsai

et al. 2023

Molecular Therapy - Methods & Clinical Development

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Section: Methodsmentioning

confidence: 99%

Partial genome content within rAAVs impacts performance in a cell assay-dependent manner

Troxell,

Jaslow,

Tsai

et al. 2023

Molecular Therapy - Methods & Clinical Development

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“…CDMS is particularly useful for characterizing very large protein complexes, − having masses that extend into the megadalton (MDa) to gigaDa regimebeyond the range of conventional MS measurements. , While investigating the formation of the larger 26S and 30S proteasomes, we found evidence for several high-mass 19S oligomers, including an abundant 19S trimer, as well as a distribution of multimeric (20S) x (where x ranges from 1 to ∼13) species, favored in acidified solutions. These larger 20S species appear to have relatively well-defined structures in which all 28 proteins comprising each core complex are entirely conserved for every oligomer size.…”

Section: Introductionmentioning

confidence: 92%

CDMS Analysis of Intact 19S, 20S, 26S, and 30S Proteasomes: Evidence for Higher-Order 20S Assemblies at a Low pH

et al. 2023

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Charge detection mass spectrometry (CDMS) was examined as a means of studying proteasomes. To this end, the following masses of the 20S, 19S, 26S, and 30S proteasomes from Saccharomyces cerevisiae (budding yeast) were measured: m(20S) = 738.8 ± 2.9 kDa, m(19S) = 926.2 ± 4.8 kDa, m(26S) = 1,637.0 ± 7.6 kDa, and m(30S) = 2,534.2 ± 10.8 kDa. Under some conditions, larger (20S) x (where x = 1 to ∼13) assemblies are observed; the 19S regulatory particle also oligomerizes, but to a lesser extent, forming (19S) x complexes (where x = 1 to 4, favoring the x = 3 trimer). The (20S) x oligomers are favored in vitro, as the pH of the solution is lowered (from 7.0 to 5.4, in a 20 mM ammonium acetate solution) and may be related to in vivo proteasome storage granules that are observed under carbon starvation. From measurements of m(20S) x (x = 1 to ∼13) species, it appears that each multimer retains all 28 proteins of the 20S complex subunit. Several types of structures that might explain the formation of (20S) x assemblies are considered. We stress that each structural type [hypothetical planar, raft-like geometries (where individual proteasomes associate through side-by-side interactions); elongated, rodlike geometries (where subunits are bound endto-end); and geometries that are roughly spherical (arising from aggregation through nonspecific subunit interactions)] is highly speculative but still interesting to consider, and a short discussion is provided. The utility of CDMS for characterizing proteasomes and related oligomers is discussed.

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“…This feature has led to a keen interest in using charge detectors to measure macro-ions that exceed the practical upper m / z limit of conventional mass analyzers. Various types of biological macro-molecules fall into this category including various organelles, viruses, − supramolecular protein complexes, − and double- and single-stranded DNA. − Charge detectors have also found interest for the measurement of nanoparticles. − …”

Section: Introductionmentioning

confidence: 99%

Charge Detection Mass Spectrometry for Megadalton Polymer Characterization and Measurement of Electrospray-Generated Charged Droplet Dynamics with a New “Direct Visualization of the Rayleigh Limit” Approach to Aid in m/z Calibration

Hrabovsky,

Argence,

Lesage

et al. 2024

Anal. Chem.

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A charge detector has been constructed and mounted inside the vacuum housing of a commercial mass spectrometer (Micromass-Waters Quattro I, Waters Corp., Manchester, UK). The in-house built single-pass charge detector is composed of a designed, complete electronics system that includes a low-noise charge amplifier. Communication to the data acquisition system was enabled, and analog and digital filters were devised, followed by their tuning and programming. Data treatment scripts for data analysis and plotting were automated, and the assembled system was calibrated and tested. The instrument has an acquisition speed of ∼200 detection events/s, and it permits detection down to ∼510 charges (= three times RMS noise) for a single measured particle. The charge detector was employed to determine the oligomer distribution of a megadalton polymer, polyethylene glycol (PEG). The PEG size distribution exhibits a maximum at ∼ m/z 5910 with the oligomeric population mass distribution peaking near 4.45 MDa. In studies of methanol droplet dynamics, "charge vs time-of-flight" plots enabled clear visualization of the zone near the Rayleigh limit to droplet charging. The highest population of methanol droplets near the Rayleigh limit carried 5000−7000 charges. This corresponds to droplet weights of 10−20 GDa, with the high-end tail extending above 70 GDa. This visualization of the most highly charged droplets (that bear numbers of charges near those defined by the Rayleigh equation) was exploited as a calibration aid for our charge detector, which lacks a means of precisely defining ion energy. A maximum m/z error of −12.3% was calculated for the method, i.e., less than the potential error in assigning the true level of charging of the most highly charged droplets relative to the Rayleigh limit. With these limitations in mind, the introduced method will provide a new means for aiding the calibration of m/z values in charge detectors.

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Charge detection mass spectrometry for the analysis of viruses and virus-like particles

Cited by 15 publications

References 64 publications

Partial genome content within rAAVs impacts performance in a cell assay-dependent manner

Partial genome content within rAAVs impacts performance in a cell assay-dependent manner

CDMS Analysis of Intact 19S, 20S, 26S, and 30S Proteasomes: Evidence for Higher-Order 20S Assemblies at a Low pH

Charge Detection Mass Spectrometry for Megadalton Polymer Characterization and Measurement of Electrospray-Generated Charged Droplet Dynamics with a New “Direct Visualization of the Rayleigh Limit” Approach to Aid in m/z Calibration

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