Scientist Solutions - Virus concentration/purification

home Posted 12/6/2010 1:07:24 PM	Can somebody please explain to me the difference between concentration of viruses and purification of viruses? What different methods can be used to obtain a concentrated virus suspension and a purified virus concentration? Centrifugation, filtration, adsorption, column chromatography, affinity chromatography, dialysis? Cheers! :)
protoldo Posted 12/7/2010 1:15:27 AM	Hola, I don´t know what virus you work, but in my case of baculovirus, purification is isolate recombinant virus from a background of wild type virus harboring betagalactosidase, because for transfection you make an homologous recombination betwen the viral DNA wt digested and the plasmid harboring the gene of the protein that you want express. About concentration it could be carried when you have an small titer and your inoculum wold be a big volume. I prefer repeat the infection to get a bigger titer but you can use ultracentrifugation ( I remember in a SW28 rotor during 1hour) or there is a method by DEAE sepharose, but in centrifugation you have to resuspend very well the pellet, and in both cases you have to keep sterility because this concentrate passage will be used to infect cells at hig multiplicity. Buena suerte
tanyag Posted 12/7/2010 4:47:47 PM	Hello Purification of virus means removing all the residual junk that is in your virus preparation. eg. cell debris, protein molecules. Purifying can be done by chromatography, centrifugation and dialysis. Concentration of virus means increasing the virus titre you have in the solution. Usually this means some kind of reduction in volume. Methods I have used are ultracentrifugation in a sucrose gradient, PEG precipitation, and Vivaflow ultrafiltration. I hope that helps.