Amplification of virus inoculum for experimental work or protein production

Amplification of virus inoculum for experimental work or protein production

As part of the service provided to make a recombinant virus, we produce a 50 ml high titre virus stock. This is then used to produce further batches of virus inoculum for protein production.

Clients may also send us their virus that they have already made and we can amplify it to high titre.  We always test the virus titre of received viruses by plaque assay before proceeding to large-scale amplification.

We can amplify virus up to 40 L in one batch.  To give an idea of the volumes of virus required for protein production the following may be useful:

Titre of virus     MOI   Cell density   Cell volume   Volume of virus required
1 x 10pfu/ml 10 2 x 106/ml 1,000ml 200ml
2 x 108 pfu/ml 10 2 x 106/ml 1,000ml 100ml
5 x 107 pfu/ml 10 2 x 106/ml 1,000ml 400ml
1 x 108 pfu/ml 10 2 x 106/ml 10,000ml 2,000ml

 

High titre virus stocks are produced using Sf9 cells in serum-free medium.  After infection, the cells are removed by centrifugation and the virus stored at 4°C in the dark.  Aliquots of virus can be prepared for long term storage.  The titre is determined by plaque-assay or baculoQUANT.

Please contact us for further details of this service