Buoyant protein harvesting method
A buoyant device containing chromatography media performs the function of protein harvesting replacing the steps of cell separation and volume reduction; the device can be loaded into columns for further purification.
1. A method of harvesting proteins in a bioreactor comprising:
a. Providing the protein-harvesting device comprising a porous container with an inner volume comprising a suitable amount of a chromatography media wherein said media is capable of binding a protein; and a buoyancy device attached to the porous container;
b. Disposing the protein-harvesting device in a container comprising a genetically modified organism or cells capable of producing a protein for harvesting in a nutrient media;
c. Adjusting the buoyancy of the protein-harvesting device to a pre-determined level;
d. Adjusting the pH and electrolyte concentration of the nutrient media to optimize the binding of the protein to the chromatography media;
e. Testing the nutrient media until the concentration of the protein in the nutrient media reaches a pre-determined low value;
f. Removing the protein-harvesting device from the nutrient media for further processing of the protein bound to the chromatography media.
2. The method of harvesting proteins according to claim 1 , wherein a plurality of protein-harvesting devices is used.
3. The method of harvesting proteins according to claim 1 , wherein the plurality of protein-harvesting devices is submerged to different depths in the container containing the nutrient media.
4. The method of harvesting proteins according to claim 1 , wherein the plurality of protein-harvesting devices contain different chromatography media or a combination of different types of chromatography media.
5. The method of harvesting proteins according to claim 1 , wherein the protein in the protein-harvesting device in step (f) is purified by:
a. Packing a plurality of protein-harvesting devices in a vertical column comprising an opening in the top and a drain in the bottom and an inner volume;
b. Adding a solution of such pH and electrolyte concentration as to break the binding between the protein and the chromatography media to recover a purified form of the protein.
6. The method according to claim 1 , wherein the buoyancy device is comprised of a continuous thin walled plastic body enclosing a substantially hollow interior.
7. The method according to claim 1 , wherein the buoyancy device is made of polymeric foam and encircling the container.
8. The method according to claim 1 , wherein the buoyancy device is made of cork and encircling the container.
9. The method according to claim 1 , wherein the buoyancy device is inflatable.
10. The method according to claim 9 , wherein the buoyancy device is inflated to different pressures to produce buoyancy.
11. The method according to claim 1 , wherein the buoyancy device additionally comprises weights to submerge it partially or completely.
12. The method according to claim 1 , wherein the container of step (b) is a flexible bag.
13. The method according to claim 1 , wherein the porous container of step (a) is made of nylon mesh or a flexible perforated plastic.
14. The method according to claim 1 , wherein the porous container of step (a) is made of perforated plastic, wood, or metal.
15. The method according to claim 1 , wherein the porous container comprises a plurality of pores having a size ranging from 5 microns to 50 microns.
16. The method according to claim 1 , wherein the porous container comprises a plurality of pores having a size ranging from 50 to 100 microns.
17. The method according to claim 1 , wherein the porous container comprises a plurality of pores having a size ranging from 100 to 300 microns.
18. The method according to claim 1 , wherein the protein harvesting device is circular, rectangular or cuboid.
19. A method of harvesting proteins in a bioreactor comprising:
a. Providing the protein-harvesting device comprising a porous container with an inner volume comprising a suitable amount of a chromatography media wherein said media is capable of binding a protein; and a buoyancy device attached to the porous container;
b. Disposing the protein-harvesting device in a container comprising a genetically modified organism or cells capable of producing a protein for harvesting in a nutrient media;
c. Adjusting the buoyancy of the protein-harvesting device to a pre-determined level;
d. Adjusting the pH and electrolyte concentration of the nutrient media to optimize the binding of the protein to the chromatography media;
e. Removing the protein-harvesting device from the nutrient media for further processing of the protein bound to the chromatography media.