Use of centrifugation-aided infection to increase virus titer
View Patent ↗The invention relates to a process for increasing the observed titer of a virus stock for the purpose of increasing the calculated log reduction (LRV) in virus clearance studies. A tissue culture or assay plate is seeded with an indicator cell line and titrated with a virus stock followed, by a centrifugation step for about 5 minutes to about 24 hours at a g-force ranging from about 50×g to about 2400×g, and at a temperature from about 4° C. to about 39° C. The resulting calculated virus titer after undergoing the centrifugation step is 10-fold higher than the virus titer would be if determined in the absence of the centrifuging step.
1. A process for increasing the observed titer of a Xenotropic murine leukemia virus (X-MuLV) stock for the purpose of increasing the calculated log reduction (LRV) in a virus clearance study comprising the steps of:
a) providing a X-MuLV stock, cell culture media, indicator cell line, and an assay plate;
b) adding the indicator cell line and media to the assay plate;
c) adding the X-MuLV stock to the cell line and media on the assay plate;
d) centrifuging the assay plate containing the cell line, media, and X-MuLV stock;
e) stopping the centrifuging step;
f) incubating the assay plate centrifuged in step d); and
g) measuring the observed titer of the X-MuLV stock, wherein the resulting titer of the X-MuLV stock is 10-fold higher than the X-MuLV titer would be if measured in the absence of the centrifuging step (d).
2. The process of claim 1 , wherein the assay plate is selected from the group consisting of a tissue culture plate, a well plate, a flask or a test tube.
3. The process of claim 1 , wherein centrifuging step (d) occurs from about 5 minutes to 24 hours at g-force range from about 50×g to 2400×g.
4. The process of claim 1 , wherein centrifuging step (d) occurs from about 30 minutes to 120 minutes at g-force range from about 500×g to 1000×g.
5. The process of claim 1 , wherein the temperature during centrifuging step (d) is about 4° C. to 39° C.
6. The process of claim 1 , wherein the temperature during centrifuging step (d) is about 21° C. to 39° C.
7. The process of claim 1 , further comprising an additional incubating step, wherein the assay plate containing X-MuLV stock, media and cell line is additionally incubated prior to the centrifuging step (d).
8. The process of claim 1 , wherein the X-MuLV stock is prediluted.
9. The process of claim 1 , wherein the incubation temperature is about 37° C.
10. The process of claim 7 , wherein the incubation temperature is about 37° C.
11. The process of claim 1 , wherein the temperature during centrifuging step (d) is about 25° C.