Hy,

attached you find a paper were we used columns quite effectively.

Sincerely Yours

Stefan

Hello Stefan,

Thank you for your reply. Have you tried this method with Adeno-associated virus? or adenovirus? or both?

Thanks again,

Deemah

Hello Deemah,

there are many papers reporting purification of AAV using columns (eg HiTrap, Mustang Q) instead of ultracentrifugation, mostly to answer the need for scalability since ultracentrifugation is not ideal for large volumes. Try looking into large scale manufacturing for more details. I personally avoid affinity columns (eg heparin) since there could be a difference in affinity between the AAV serotypes. If you want a rough but quick solution you can try some AAV purification kits.

Here's a review that might help you:

Merten, O.-W., Schweizer, M., Chahal, P. & Kamen, A. Manufacturing of viral vectors: part II. Downstream processing and safety aspects. Pharmaceutical Bioprocessing 2, 237–251 (2014).

About the second question, you can achieve large production of virus without purification but it goes without saying that the product will be contaminated by cellular material. For example, if you spin the cell lysate with a 100kDa centrifugal filter to concentrate the virus you will have lot of cell debris with it, even after clarification of cell lysate with low speed centrifugation or 0.2um filtration.

Hope this helps

Davide

Thank you Davide.