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Purification of Murine Gammaherpesvirus 68 With Use of Differential Centrifugation

R. Hodoši

R. Hodoši

Department of Microbiology and Virology, Faculty of Natural Sciences, Comenius University in BratislavaBratislava, Slovak republic
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Hodoši, R.
, 
E. Nováková

E. Nováková

  • Department of Microbiology and Virology, Faculty of Natural Sciences, Comenius University in BratislavaBratislava, Slovak republic
  • Slovak Academy of Sciences, Biomedical Research Center, Institute of VirologyBratislava, Slovak republic
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Nováková, E.
 and 
M. Šupolíková

M. Šupolíková

  • Department of Microbiology and Virology, Faculty of Natural Sciences, Comenius University in BratislavaBratislava, Slovak republic
  • Department of Galenic Pharmacy, Faculty of Pharmacy, Comenius University in BratislavaBratislava 3, Slovak Republic
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Šupolíková, M.
  
Jul 30, 2021
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Article Category: Special Issue Article
Published Online: Jul 30, 2021
Page range: 76 - 79
Received: Jun 14, 2021
Accepted: Jun 17, 2021
DOI: https://doi.org/10.2478/afpuc-2021-0009
Keywords
© 2021 R. Hodoši et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Figure 1

Diagram of particle behaviour during differential centrifugation and density gradient centrifugation. a) During the differential centrifugation of a suspension of particles in a centrifugal field, the movement of the particles depends on their density, shape and size. b) To separate the biological particles by means of a density gradient, the samples are layered on the prepared gradient before centrifugation. To achieve separation, the sample is centrifuged, until the isopycnic position for the desired particles in the gradient is achieved.
Diagram of particle behaviour during differential centrifugation and density gradient centrifugation. a) During the differential centrifugation of a suspension of particles in a centrifugal field, the movement of the particles depends on their density, shape and size. b) To separate the biological particles by means of a density gradient, the samples are layered on the prepared gradient before centrifugation. To achieve separation, the sample is centrifuged, until the isopycnic position for the desired particles in the gradient is achieved.

Figure 2

Schematic diagram of virus purification steps.
Schematic diagram of virus purification steps.

Figure 3

Polypeptide profile of purified MHV-68 in gradient SDS-PAGE (6%–14%, w/v). Lane 1: ladder (Precision Plus Protein™ Dual Xtra); lane 2: purified MHV-68 (centrifuged 6 hours) – 10 μl/lane; lane 3: purified MHV-68 (centrifuged 6 hours) – 5 μl/lane; lane 4: purified MHV-68 (centrifuged 6 hours) – 3 μl/lane.
Polypeptide profile of purified MHV-68 in gradient SDS-PAGE (6%–14%, w/v). Lane 1: ladder (Precision Plus Protein™ Dual Xtra); lane 2: purified MHV-68 (centrifuged 6 hours) – 10 μl/lane; lane 3: purified MHV-68 (centrifuged 6 hours) – 5 μl/lane; lane 4: purified MHV-68 (centrifuged 6 hours) – 3 μl/lane.

Figure 4

Molecular weights (in kDa) of viral proteins of purified MHV-68, gradient SDS-PAGE (6%–14% w/v). Lane 1: ladder (Precision Plus Protein™ Dual Xtra); lane 2: purified MHV-68 (centrifuged 6 hours) – 12 μl/lane; lane 3: purified MHV-68 (centrifuged 6 hours) – 6 μl/lane.
Molecular weights (in kDa) of viral proteins of purified MHV-68, gradient SDS-PAGE (6%–14% w/v). Lane 1: ladder (Precision Plus Protein™ Dual Xtra); lane 2: purified MHV-68 (centrifuged 6 hours) – 12 μl/lane; lane 3: purified MHV-68 (centrifuged 6 hours) – 6 μl/lane.
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English
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Pharmacy, Pharmacy, other
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