Downstream bioprocessing device

An expanded-bed adsorption chromatography method for purifying a target protein in a protein solution comprising:

• a. providing a chromatography column comprising a cylinder capable of holding a liquid wherein the cylinder comprises:
• i. a top opening comprising a top filter covering a top opening and a removable top cap to hold the top filter in place comprising at least one top liquid port and a top flow control valve;
• ii. a top sampling port comprising a top sampling valve connected to the top liquid port;
• iii. a bottom opening comprising a bottom filter covering the bottom opening and a removable bottom cap to hold the bottom filter disk in place comprising a bottom liquid port, a bottom flow control valve;
• iv. a bottom sampling port comprising a bottom sampling valve connected to the bottom liquid port;
• v. a means of mixing the contents of the cylinder;
• and
• vi. a plurality of sensors to disposed inside the cylinder to measure turbidity of the target protein solution in the cylinder.
• b. connecting the bottom liquid to a source of a target protein solution with approximate known target protein content, in need for purification;
• c. removing the top cap and adding to the cylinder a quantity of a chromatography media sufficient to bind substantially all of the target protein content in the target protein solution;
• d. replacing the top cap and opening the top and bottom flow control valves;
• e. filling the cylinder with the target protein solution under gravity or through peristaltic pumps through the bottom liquid port;
• f. closing the bottom flow control valve;
• g. mixing the chromatography media to achieve a uniform distribution in the cylinder as indicated by the measurement of turbidity by the sensors and adjusting the speed of mixing to achieve a uniform turbidity;
• h. connecting each of the top and bottom sampling ports to a flow cell of a spectrophotometer capable of measuring the concentration of the target protein and turning on the bottom flow control valve when the ratio of the concentration in the top and bottom sampling ports reaches about 1:100 and closing the bottom flow control valve when the ratio reaches about 2:100;
• i. maintaining the flow of target protein solution into the cylinder and allowing the target protein solution to flow out of the top liquid port;
• j. closing the bottom flow control valve and the liquid ports and disconnecting the source of target protein solution;
• k. opening the bottom flow control valve to allow the target protein solution in the cylinder to drain out through bottom liquid port;
• l. connecting the bottom flow control port to a source of an elution buffer capable of breaking the binding of the target protein to the chromatography media;
• m. filling the cylinder with elution buffer;
• n. closing the bottom flow control valve and begin mixing;
• o. continue mixing the contents of the cylinder for a desired time to allow complete breaking of the binding between the target protein and the chromatography media;
• p. stopping the mixing;
• q. opening the bottom flow control valve and the liquid ports and collecting the elution buffer in a container as a purified concentrated solution of the target protein.