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Concentration and lysis of adenovirus-infected cells in a single unit operation
| Details |
Inventors: Monica, Thomas J.; Whiteley, Erik M.;
Assignee: Schering Aktiengesellschaft (Berlin, DE)
Primary Examiner: Housel; James
Assistant Examiner: Chen; Stacy B
Attorney, Agent or Firm: Millen, White, Zelano & Branigan, P.C.
A method is described to prepare intracellular biological entities (e.g. organisms, such as, e.g., viruses, particularly Adenoviruses; organelles, or biological molecules), comprising subjecting cells containing the biological entities to continuous centrifugation, under conditions effective to concentrate the cells into a cell pellet; and ejecting the pelleted cells from the centrifuge into a collection receptacle, under conditions effective to lyse cells; wherein no further steps effective to achieve cell lysis are performed. |
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DETAILED DESCRIPTION What is claimed is: 1.
A method to release viruses from animal cells containing said viruses, comprising subjecting the cells to continuous centrifugation under conditions effective to concentrate the cells into a cell pellet, and ejecting the pelleted cells from the centrifuge through one or more ejection outlets into a collection receptacle, under conditions effective to lyse said cells solely by exertion of forces generated by said centrifugation and to release said viruses from said cells.
2.
The method of claim 1, wherein greater than about 85% of the cells are lysed.
3.
The method of claim 1, wherein greater than 50% of the cells are lysed.
4.
The method of claim 1, wherein the cells are lysed as they are ejected.
5.
The method of claim 1, wherein the viruses are Adenoviruses.
6.
The method of claim 5, wherein the Adenoviruses are recombinant Adenoviruses suitable for gene therapy.
7.
The method of claim 5, wherein the yield/cell of Adenovirus particles or infectious Adenovirus is greater than that obtainable when cells containing said Adenovirus are lysed by a freeze-thaw procedure.
8.
The method of claim 7, wherein the yield/cell of Adenovirus particles is about 1.
2 to about 1.
6 fold greater than that obtainable when cells containing said Adenovirus are lysed by a freeze-thaw procedure.
9.
The method of claim 7, wherein the yield/cell of infectious Adenovirus is about 1.
5 to about 1.
9 fold greater than that obtainable when cells containing said Adenovirus are lysed by a freeze-thaw procedure.
10.
The method of claim 1, wherein the cells are mammalian or insect cells.
11.
The method of claim 1, wherein the cells being ejected are under a relative centrifugal force of 6500 to 7500 g.
12.
The method of claim 11, wherein said centrifugal force is about 7000 g.
13.
The method of claim 1, wherein the pelleted cells are ejected through one or more ejection outlets having a rectangular shape and a cross-sectional area of 50 to 500 mm.
sup.
2.
14.
The method of claim 11, further wherein the pelleted cells are ejected through one or more ejection outlets having a rectangular shape and a cross-sectional area of 50 to 500 mm
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Molecular Biology 
