HARVESTING AND PERFUSION APPARATUS

1 . An apparatus to harvest expressed protein from culture media comprising:

a. A first container with inner volume, a top surface, and a bottom surface;

b. A first liquid port in the bottom surface and comprising a valve or clamp to open or close the first liquid port for inlet and outlet of liquid;

c. A second liquid port at the bottom surface with a valve or clamp to open or close the second liquid port in the first container for the outlet of liquid;

d. A third liquid port at the top surface comprising a valve or a clamp to close or open the third liquid port for inlet of liquid and an optional filter;

e. A first gas port at the top surface comprising

i. A valve or a clamp to close or open the first gas port for inlet of gas;

ii. A sparging rod disposed at the bottom of the first container; and

iii. an optional filter;

f. A second gas port at the top surface comprising a valve or a clamp to close or open the second gas port for outlet of gas and an optional filter;

g. A second container with an inner volume comprised of a porous material containing a binding resin, wherein the second container is disposed inside the first container;

h. A flexible tube connecting the first liquid port in bottom surface of the first container to a bioreactor; and

i. A movable platform to support the first container capable of moving the first container up and down.

2 . The apparatus of claim 1 , further comprising a weight sensor attached to the movable platform.

3 . The apparatus of claim 2 , wherein the weight sensor is connected to a device capable of automatically controlling the movable platform to move up or down based on a pre-determined weight limit of the first container.

4 . The apparatus of claim 1 , further comprising a filter disposed inside the first container and attached to the first liquid port and a vibrator attached to the filter attached to the first liquid port.

5 . The apparatus of claim 1 , where the first container is a flexible bag.

6 . The apparatus of claim 1 , wherein the second container is a flexible pouch.

7 . The apparatus of claim 1 , wherein the second container is buoyant.

8 . The apparatus of claim 1 , wherein the second container is comprised of nylon mesh, a flexible perforated plastic, wood or metal.

9 . The apparatus of claim 1 , wherein the second container comprises a plurality of pores having a size ranging from 5 microns to 50 microns.

10 . The apparatus of claim 1 , wherein the second container comprises a plurality of pores having a size ranging from 50 to 100 microns.

11 . The apparatus of claim 1 , wherein the second container comprises a plurality of pores having a size ranging from 100 to 300 microns.

12 . The apparatus of claim 1 , wherein the binding resin is Protein A.

13 . The apparatus of claim 1 , wherein the binding resin is a mixed bed resin.

14 . The apparatus of claim 13 , wherein the resin is selected from ion exchange resins, hydrophobic chromatography resins, and affinity resins.

15 . A method of harvesting an expressed protein from a culture media produced in a bioreactor comprising:

a. Providing the apparatus of claim 1 ;

b. Connecting the first liquid port to a bioreactor wherein the protein is produced;

c. Opening the first gas and second gas ports;

d. Starting flow of gas through first gas port to begin gas flow through the sparging tube;

e. Lowering the platform supporting the apparatus to below the level of the bioreactor;

f. Allowing gravity flow of culture media in the bioreactor into the first container;

g. Raising the platform supporting the apparatus when the weight of the apparatus reaches a predetermined level, to above the level of the bioreactor;

h. Allowing gravity flow of the culture media from the first container back into the bioreactor;

i. Repeat steps (e) to (h) for a pre-determined number of times based on binding capacity of the resin disposed of in the second porous container;

j. Raising the platform to a level above the level of the bioreactor and allowing culture medium in the first container drain into the bioreactor;

k. Closing the first liquid port between the bioreactor and the apparatus;

l. Opening the second liquid port and introducing an eluting liquid capable of eluting the protein from the resin in the second porous container;

m. Opening the third liquid port and draining the eluting liquid from the apparatus for further processing;

n. Closing the third liquid port;

o. Introducing a washing liquid through the second liquid port into the apparatus;

P. Allowing the washing liquid to sit for a period of time;

q. Opening the third liquid port and draining the washing liquid from the apparatus;

r. Repeat steps (p) to (r) for a pre-determined number of times and discarding the washing liquid;

s. Closing the third liquid port;

t. Opening the first liquid port; and

u. Repeating steps (e) to (t) for a pre-determined number of times to harvest part or all of the protein produced in the bioreactor.

16 . The method of claim 15 , wherein the method is used at the end of a protein expression cycle in the bioreactor.

17 . The method of claim 15 , wherein the method is used during the production of the protein in the bioreactor.

18 . The method of claim 15 , wherein the said method is used continuously or intermittently during the production of the protein in the bioreactor.

19 . The method of claim 15 , wherein the binding of the protein to resin stabilizes the protein, thereby improving yield.

20 . The method of claim 15 , wherein the binding of the protein to resin improves the yield of production due to removing a toxic protein from the media.

21 . A method for refolding proteins comprising:

a. Obtaining a purified protein in need of refolding;

b. Diluting the protein in a refolding buffer;

c. Introducing the protein in said buffer into the apparatus of claim 1 through the second liquid port;

d. Allowing the protein to bind to the resin in the second porous container;

e. Opening the third liquid port and draining the refolding buffer from the apparatus;

f. Closing the third liquid port;

g. Introducing a washing liquid through the second liquid port into the apparatus;

h. Allowing the washing liquid to sit for a period of time;

i. Opening the third liquid port and draining the washing liquid from the apparatus;

j. Repeat steps (g) to (i) for a pre-determined number of times and discarding the washing liquid;

k. Opening the second liquid port and introducing an eluting liquid capable of eluting the refolded protein from the resin;

l. Opening the third liquid port and draining the eluting liquid from the apparatus for further processing;

m. Closing the third liquid port.