Question

Describe a purification scheme that starts with a mixture of cells, virions and proteins and ends with a tube containing pure virions. Or, how about obtaining a supernatant solution containing only pure virions?

Answer 1

Both enveloped and non-enveloped virions can be purified using some very useful techniques such as Centrifugation, chromatography etc.

To start with the mixed culture, virios need a host in order to survive within. Hence the virions can be subjected to the supernatant by breaking the cell membrane and releasing the virions.

1. Centrifugation of the mixture prior to addition of cell-disruptors like any enzyme(eg.lysosyme)

2. Discarding the pellet containing the disrupted cells and collecting the supernatant in a different vial

3. This supernatant contains a mixture of proteins and viruses that can be now be subjected to ultracentrifugation of chromatography

4. Ultracentrifugation at higher rpm will separate macromolecules like the proteins from the mixture

5. The proteins will be pelleted down due to it's heavy molecular weight and the smaller virion particles might remian in the supernatant

6. Collect the virion containing supernatant in a fresh vial