COMPOSITIONS AND METHODS FOR REDUCING CHROMATIN CONTENT OF BIOLOGICAL PREPARATIONS
A method for removing chromatin from a cell culture harvest comprising the steps: providing a cell culture harvest containing a desired biological product selected from the group consisting of a parvovirus or an adeno-associated virus, incubating the cell culture harvest in an aqueous medium at a pH value within the range of 3.0 to 4.0 with an ionic strength corresponding to NaCl at a concentration within the range of 3.0 M to saturation and separating the desired biological product from solids produced.
1 . A method for removing chromatin from a cell culture harvest comprising the steps:
providing a cell culture harvest containing a desired biological product selected from the group consisting of a parvovirus or an adeno-associated virus,
incubating the cell culture harvest in an aqueous medium at a pH value within the range of 3.0 to 4.0 with an ionic strength corresponding to NaCl at a concentration within the range of 3.0 M to saturation and separating the desired biological product from solids produced.
2 . The method of claim 1 wherein the cell culture harvest is obtained from a culture of mammalian cells, insect cells, yeast cells, or bacterial cells.
3 . The method of claim 1 wherein the aqueous medium comprises sodium chloride at a concentration in the range of 3.0 M up to saturation.
4 . The method of claim 1 wherein the pH is 3.5±0.3.
5 . The method of claim 1 wherein incubating is performed for up to 24 hours.
6 . The method of claim 1 wherein the solids are separated from the soluble product by sedimentation or filtration.
7 . The method of claim 1 wherein incubation incubating is performed in the presence of a solid-phase material bearing primary amine groups.
8 . The method of claim 7 wherein the primary amine-bearing solid phase in the form of loose particles or in the form of a chromatography device.
9 . The method of claim 7 wherein solid phase particles are present in a volumetric proportion in the range of 1% to 10% relative to the volume of cell culture harvest in an aqueous medium.
10 . The method of claim 7 wherein the primary amine-bearing solid phase is in a form selected from the group consisting of columns packed with porous particles, columns packed with nanofibers, columns packed with monolithic materials, hydrogels, porous membrane filters, and depth filters.
11 . The method of claim 7 , wherein the primary amine surface of the solid phase is accompanied by chemical residues of alternative character, for example secondary amines, tertiary amines, quaternary amines, hydrophobic residues, hydrogen bonding residues, metal affinity residues, or combinations thereof.
12 . The method of claim 1 , wherein the method is performed in the presence of one or more salts, sugars, surfactants, divalent metal ions, allantoin, polyethylene glycols, flocculating agents, or combinations thereof.
13 . The method of claim 1 , further comprising the steps of
reducing ionic strength of a solution comprising the desired biological product
removing further contaminants by contacting the solution with a cation exchanger, wherein the cation exchanger binds the contaminants
further reducing ionic strength of the solution comprising the desired biological product
purifying the desired biological product by contacting the solution with a cation exchanger, wherein the cation exchanger binds the desired biological product.
14 . (canceled)
15 . The method of claim 1 wherein incubating is performed for 45 minutes to 90 minutes.