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Virus vs protein purification


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#1 saku98

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Posted 11 February 2020 - 07:21 AM

I am looking to purify virus that is incorporated with a protein (a luminescence tag).

So I collected supernatant of infected cells. The virus containing supernatant I then centrifuged at max speed for 90min and discard the supernatant. The pelleted virion I resuspend in PBS and detect the protein using luminescence substrate.

 

My question is, when I centrifuge as in above condition, can I expect the pelleted is indeed just virion, and any other protein that was expressed by the cells should be in the supernatant after centrifugation. Protein weight much lighter and should not be able to pellet without any precipitation strategy right?

 

 

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#2 mdfenko

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Posted 12 February 2020 - 01:54 PM

There may still be some protein in the pellet. The rate at which protein will pellet is determined by the sedimentation coefficient, viscosity of the medium and rcf. It shouldn’t pellet all at once, but will gradually come down into the pellet. The virion will just pellet faster due to its greater sedimentation coefficient.


Edited by mdfenko, 12 February 2020 - 02:01 PM.

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#3 jamesholder

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Posted 29 November 2020 - 11:43 PM

Due to the already reported problems of virus isolation and purification from garlic, serology is not a very good tool for virus species differentiation among allexiviruses. However, polyclonal antibodies were raised against GMbMV (Yamashita et al., 1996), which was actually proved to be GarV-C (Tsuneyoshi and Sumi, 1996; Yamashita et al., 1996), and GarV-D (E. Barg, personal communication). Polyclonal and/or monoclonal antisera have also been produced against ShVX, GarV-A, GarV-B, and GarV-C (D. E. Lesemann and E. Barg, personal communication).






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