Method for generating and purifying viral vectors
OwnerMartin Saballus, Markus Kampmanninventors
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Abstract
The invention relates to a method for generating and purifying viral vectors (V), wherein the viral vectors (V) are generated by cells in a fluid (F) and wherein the fluid (F) comprising the viral vectors (V) is guided through a tangential flow filtration arrangement (2) for purification, whereby the viral vectors (V) are purified in the fluid (F). It is proposed that the cells and cell debris from the cells, which are contained in the fluid (F) comprising the viral vectors (V) after generating the viral vectors (V), are at least partially separated from the viral vectors (V) by a fluidized bed centrifuge (5) before purification by the tangential flow filtration arrangement (2).
Claims14
Method for generating and purifying viral vectors (V), wherein the viral vectors (V) are generated by cells in a fluid (F) and wherein the fluid (F) comprising the viral vectors (V) is guided through a tangential flow filtration arrangement (2) for purification, whereby the viral vectors (V) are purified in the fluid (F),
- characterized in
- that the cells and cell debris from the cells, which are contained in the fluid (F) comprising the viral vectors (V) after generating the viral vectors (V), are at least partially separated from the viral vectors (V) by a fluidized bed centrifuge (5) before purification by the tangential flow filtration arrangement (2).
Method according to one of the previous claims, characterized in that the fluid (F) comprising the viral vectors (V) is guided from the fluidized bed centrifuge (5) into an intermediate holding vessel (13), preferably in that the fluid (F) flows into the intermediate holding vessel (13) discontinuously and flows out of the intermediate holding vessel (13) continuously.
Method according to one of the previous claims, characterized in that enzymes are added to the fluid (F) so that the cells and/or the cell debris and/or the DNA impurities are degraded, in particular thereby releasing the viral vectors (V), and/or reduced in molecular size, preferably, that the enzymes are added to the fluid after the fluidized bed centrifuge (5) and/or prior to the tangential flow filtration arrangement (2).
Method according to one of the previous claims, characterized in that the viral vectors (V) are concentrated in the fluid (F), which flows out of the tangential flow filtration arrangement (2) as retentate (R) and impurities are separated into the fluid, which flows out of the tangential flow filtration arrangement (2) as permeate (P).
Method according to one of the previous claims, characterized in that the fluid (F) comprising the viral vectors (V) is guided through an ion exchange arrangement (15), preferably after the tangential flow filtration arrangement (2), and the viral vectors (V) are purified.
Method according to one of the previous claims, characterized in that the fluid (F) in a initial holding vessel (12) and/or in the fluidized bed centrifuge (5) and/or in the intermediate holding vessel (13) and/or in the tangential flow filtration arrangement (2) and/or in the ion exchange arrangement (15) and/or in fluid lines (18) for guiding the fluid (F) is at least partially cooled and/or heated.
System for generating and purifying viral vectors (V), in particular for performing the method according to one of the previous claims, comprising a tangential flow filtration arrangement (2) for purification of the viral vectors (V) generated by cells in a fluid (F),
- characterized in
- that the system (1) comprises a fluidized bed centrifuge (2) for separating the cells and/or cell debris from the cells contained in the fluid (F) after generating the viral vectors (V) and prior to purification by the tangential flow filtration arrangement (2).
Dependencies are inferred from the claim text.