virus purification from tissue

Yersinia yersinia at CYBERNEX.NET
Sun Nov 23 12:23:42 EST 1997


Chien-Chang Loa writes,

<I am trying to purify coronavirus from intestinal content. However, it 
is so hard to get rid of the host proteins.
Any suggestions?>

I'll admit I'm taking a big chance here (of making a complete fool of 
myself) - I'm only a virology student this semester, I'm not an actual 
virologist.  But your question sounds simple enough, so I'll give it a 
try. In our labs, we had to grow up two types of viruses and purify them 
for use in quantitative studies. Though they weren't coronaviruses, here 
is what we did:

A.  T4 Phage: We had to grow them in E. coli B, and then make a pure 
virus suspension - we centrifuged the tube of virus-infected bacteria at 
4500 rpm for 15 minutes. Then we decanted the supernatant, which was our 
suspension of pure virus, later used in a phage-titer experiment.

B. Poliovirus Type 3: We grew these viruses in HEp-2 cells in tissue 
culture, and harvested the pure virus by centrifuging the tube of 
infected cells at 2000 rpm for 10 minutes, then decanted the supernatant, 
which we successfully used for an infectivity titration.

I would imagine (and someone please correct me if I err or otherwise 
oversimplify!) that you could get a pure coronavirus suspension by 
centrifuging your intestinal cells and decanting the supernatant.

Happy Hunting!

Infectionately,
Yersinia.

"Hell no, let 'em grow!"




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