Characterization of Purified Ms2 Bacteriophage by the Physical Counting Methodology Used in the Integrated Virus Detection System (Ivds)

Abstract. Characterizing supramolecular interactions offers significant challenges using NMR or crystallographic techniques either because of size limitations or the difficulty in forming suitable crystals, while mass spectrometry is largely limited to low resolution mass information. Here we report gas phase measurements of intact virus particles using electrospray ion mobility spectrometry with an accuracy in radial measurements that were sufficient to differentiate closely related species. In addition, measured diameters indicate that iscosahedral virus particles retain their structure in the gas phase as well as undergoing a slight compaction in the absence of solvent. Analysis of the human pathogen adenovirus represents the largest and most sophisticated biomolecular complex detected in the gas phase to date. These results, on a diverse set of viral systems, suggest that ion mobility spectrometry may have broad applications for the analysis of biological complexes.

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“…The samples were dialyzed against 25 mM ammonium acetate at acidic (2.8) and neutral (7.4) pH. MS2 was purified as previously described [18].…”

Section: Methodsmentioning

confidence: 99%

“…A recent study successfully compared the diameter of 32 monomeric and multimeric globular proteins based on their ion mobility [17] and demonstrated the potential of IMS for studying intact virus particles. A second IMS study reported that the modal diameter of MS2 particles could be measured using IMS [18].…”

Section: Introductionmentioning

confidence: 99%

Electrospray ion mobility spectrometry of intact viruses

Thomas

Bothner

Traina³

et al. 2004

Journal of Spectroscopy

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“…As mentioned in the previous section, the first reported application of ES-DMA to biological molecules was by Kaufman et al in 1996 for characterizing proteins. This work was followed over the next decade by several reports from Allmaier et al [11], Wick et al [12] and de la Mora et al [13] related to characterization of several other proteins, viruses and polymers. Starting in 2006 as seen in Figures 1.3A and 1.3B a dramatic increase in the number of publications and citations related to ES-DMA occurred with contributions from Biswas et al [14], Zachariah et al [15], Loo et al [16], Hogan et al [17], Pergantis et al [18], Pease et al [19] and Hackley et al [20].…”

Section: Growth Of Es-dmamentioning

confidence: 98%

Electrospray–differential mobility analysis of bionanoparticles

Guha

Tarlov

et al. 2012

Trends in Biotechnology

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The growth of the multibillion dollar bionanoparticle industry has spurred the development of new physical characterization methods. One such method, electrospraydifferential mobility analysis (ES-DMA) constitutes an electrospray for aerosolization of bionanoparticles (such as viruses, gold-nanoparticles, proteins, nanoparticle-protein complexes) and an ion mobility method that operates at atmospheric conditions, and separates bionanoparticles spatially. This dissertation identifies some relevant "problem" areas for ES-DMA by reviewing selected applications.Some such problems are: proteins while passing through ES capillaries are found to interact with it and thus produce time dependent size distributions. Further, it is thought that adsorbed proteins may subsequently desorb and influence size distributions with the ES-DMA which may concomitantly affect quantification of aggregates. These artifacts are studied systematically and it is demonstrated that ES-DMA can quantify adsorption-desorption of complex protein mixtures at high shear rates. Further, it is shown that desorbing proteins do not have a significant effect on size distributions. Another artifact of the ES takes place during the aersolization process. Two units (called monomers) of a bionanoparticle may get encapsulated in the same ES droplet and upon drying of the droplet create artificial dimers thus affecting quantification with ES-DMA. Assuming Poisson distribution, this thesis provides a systematic approach that can be undertaken to eliminate this artifact. A third artifact arises from the low sensitivity of the DMA to size increase. When a ligand (for e.g. protein) adsorbs to a bionanoparticle it creates an increase in the size of the later, which can be used to quantify the amount of ligand adsorbed per bionanoparticle. As ligands can change conformations upon adsorption, using ES-DMA for such applications may be flawed. This issue has been identified and a solution has been provided by integrating a mass analyzer after the ES-DMA.After correcting for these artifacts, this dissertation delves into characterization of different types of bionanoparticles and demonstrates that ES-DMA has several advantages over other traditional techniques such as transmission electron microscopy, size exclusion chromatography, analytical ultracentrifugation, dynamic light scattering and plaque assay and thus has immense potential to become a process analytical technique in biomanufacturing environments. ELECTROSPRAY-DIFFERENTIAL MOBILITY ANALYSIS OF BIONANOPARTICLES

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“…ET AL. particles have been studied as a means of developing a calibration curve for estimating the molecular masses of large organic complexes (Bacher et al 2001) and to develop an integrated virus detection system (IVDS) for estimating virus particle concentrations and virus particle agglomeration in liquid systems (Wick and McCubbin 1999a). While virus capsid agglomeration has been documented in several studies (Thomas et al 1998;Wick and McCubbin 1999a), previous studies of airborne virus particles have employed the use of electrospray to allow for only single-virus particle aerosolization (Bacher et al 2001;Wick and McCubbin 1999a). Because of their small size, virus particles have a high deposition efficiency within the alveoli of mammalian respiratory systems and can lead to a variety of medical illnesses.…”

Section: Introductionmentioning

confidence: 99%

Capture of Viral Particles in Soft X-Ray–Enhanced Corona Systems: Charge Distribution and Transport Characteristics

Hogan

Lee

Biswas

2004

Aerosol Science and Technology

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The charge distribution of airborne MS2 bacteriophage nanoparticles and the efficiency of electrical-mobility-based capture mechanisms with bipolar charging were studied. MS2 virions form large agglomerated particles in a suspension. The average charge on airborne MS2 virions can be as high as one unit charge (negatively charged). The application of both soft X-ray irradiation and alpha rays from a Po-210 bipolar charger was shown to not only reduce the average charge on MS2 virion particles but also partially fragment the larger MS2 virion agglomerates, thereby increasing the number of ultrafine MS2 virion particles. A cylindrical electrostatic precipitator with a mounted soft X-ray emitter was used to determine the effectiveness of electrical capture methods for virus particles. At low applied voltages, it was found that the capture efficiency of ultrafine virus particles can be increased by applying in situ soft X-ray irradiation with electrostatic precipitation. It has also been shown that in the presence of both a positive and negative corona, virus particles are readily captured with log removal values exceeding 4. The unit developed and demonstrated in this work is a compact, low-pressure drop system that can be readily mounted in ventilation ducts or air supply systems to remove ultrafine particles such as viruses.

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Characterization of Purified Ms2 Bacteriophage by the Physical Counting Methodology Used in the Integrated Virus Detection System (Ivds)

Cited by 37 publications

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Electrospray ion mobility spectrometry of intact viruses

Electrospray ion mobility spectrometry of intact viruses

Electrospray–differential mobility analysis of bionanoparticles

Capture of Viral Particles in Soft X-Ray–Enhanced Corona Systems: Charge Distribution and Transport Characteristics

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