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Bioprocess engineering, also called biochemical engineering, is a specialization of chemical engineering or biological engineering, dealing with the design and development of equipment and processes for the manufacturing of products such as agriculture, food, feed, pharmaceuticals, nutraceuticals, chemicals, and polymers and paper from biological materials. The basic techniques of bioprocessing data back thousands of years with manufacture of beer, yogurt and bread. In the last few decades, bioprocess engineering has expanded into manufacturing of highly active drugs, chemicals, and organs bringing high scrutiny of processes from the regulators worldwide. The inventions listed below deal with several key steps in bioprocess engineering, from upstream (growth of a living entity) to downstream (harvesting and purification), to monitoring and making bioreactors more efficient, able to produce affordable drugs and other products.

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Upstream Systems

Current bioreactors date back to thousands of years and even in their modern structure to over 180 years; the inventions described below reinvent the design of bioreactors from disposing them horizontally instead of vertically, making them disposable, providing an orbital motion, improving gasification, eliminating need to operate them in clean rooms and many more functions that make them more cost-effective, more useful in manufacturing modern biological drugs and allow development by smaller entities.

US 9,550,971 | Universal bioreactors and methods of use.

Bioreactors suitable for housing a predetermined volume of culture medium comprising a plurality of containers of approximately equal internal volume in which the culture medium resides, wherein the predetermined volume of culture medium is retained in approximately equal amounts within each of the plurality of containers during operation of the bioreactor, and wherein the internal volume of each container is selected so that the amount of culture medium in each container exceeds 50 vol. % of the container internal volume during operation of the bioreactor, and related methods of use. A method for the continuous preparation of a biological product from a culture medium comprising: (a) providing a bioreactor system comprising: (i) a plurality of vertically disposed disposable containers for retaining the culture medium, the plurality of containers comprising a first disposable container disposed at the highest level, a last disposable container disposed at the lowest level and an intermediate disposable container disposed intermediate to the first and last containers, each container comprising: at least one culture medium inlet, at least one culture medium outlet separate from the at least one culture medium inlet, wherein fluid can flow out of the at least one culture medium outlet at the same time that fluid flows into the at least one culture medium inlet, at least one gas inlet, at least one sparging filter attached to the at least one gas inlet, the sparging filter comprising a plurality of pores on at least a portion thereof, the portion of the filter comprising the pores extending into the culture medium, at least one gas exhaust, at least one foam trap attached to the at least one gas exhaust, at least one inlet in fluid communication with the foam trap, one or more sensors for sensing one or more parameters of the culture medium, (ii) a flow conduit comprising a valve which permits the flow of culture medium under gravity through the at least one culture medium outlet of a container and into a subsequent container through the at least one culture medium inlet thereof, (iii) at least one selectively movable base which supports each container, wherein movement of the base induces movement of the culture medium within each container, (b) charging the first container with culture medium comprising a component selected from the group consisting of mammalian cells, plant cells and bacteria, (c) moving the culture medium in the first container while introducing oxygen into the medium through the sparging filter until the desired density of the cells in the culture medium is obtained, (d) opening the valve in the flow conduit located between the first container and the intermediate container to permit the culture medium from the first container to enter the intermediate container continuously at a pre-determined rate, (e) charging continuously the first container with a culture medium comprising the component included in the culture medium of step b to make up the volume of the culture medium transferred to the intermediate container, and continue moving the culture medium in the first container, and adding culture medium to the cell culture medium present in the intermediate container to a pre-determined volume, (f) continuously moving the culture medium in intermediate container while introducing oxygen into the medium through the sparging filter until the desired density of the cells in the culture medium of the intermediate container is obtained, (g) opening the valve in the flow conduit located between an intermediate container and the last container to permit the culture medium from the intermediate container to enter the last container, continuously at a pre-determined rate, (h) charging continuously the intermediate container with a culture medium to replace the volume transferred to the last container, and adding culture medium to the culture medium present in the last container to a pre-determined volume, (i) moving the culture medium in the last container while introducing oxygen into each medium through the sparging filter until the desired density of the cells in the culture medium in the last container is obtained, (j) removing the culture medium continuously from the last container by opening the valve in the flow conduit in last container and collecting the culture medium in a receptacle for further processing, and continuously adding culture medium to the culture medium present in the last container to maintain the pre-determined volume of the culture medium in the last container. PDF.

US 9,500,381 | Multiuse Reactors and Related Methods.

A septum is positioned within a disposable vessel and defines a lower chamber and an upper chamber. The septum includes a plurality of apertures that provide fluid communication between the upper chamber and lower chamber. Compressed gas is introduced in the lower chamber to produce fine bubbles rising throughout the vessel to produce a mixing and gasification needed for the growth of a biological culture and manufacture of a biological product in a nutrient medium. Adding a binding resin to the upper chamber allows harvesting, separation and purification of biological products in the reactor as a single unit operation. A reactor capable of holding liquid comprising: a) A flexible disposable bag comprising a top layer, a bottom layer, and a perforated sparging middle layer held in close proximity to the bottom layer by a plurality of spot-welds between the bottom layer and the middle layer; b) At least one liquid inlet; and at least one liquid outlet; c) At least one gas outlet connected to the top layer d) At least one gas inlet connected to the bottom layer and further connected to a source of compressed gas; e) At least one sampling port; f) An outer support to hold the bag, additionally comprising a heating and/or cooling element. PDF

US 9,499,290 | Stationary Bubble Reactors.

 Reactors that allow mixing and gasification by converting the entire floor of the reactor vessel to a sparge filter is described. A flexible horizontally disposed disposable bioreactor bag capable of holding nutrient media comprising: a flexible top sheet, a porous flexible middle sheet, and a flexible bottom sheet, the porous flexible middle sheet tufted to the flexible bottom sheet to form a sparging compartment covering essentially an entire bottom surface of the bioreactor bag, wherein the flexible top sheet and the flexible bottom sheet are sealed together at their edges and the sparging compartment serves to aerate and mix nutrient media located in a nutrient media compartment formed between the flexible top sheet and the porous flexible middle sheet; a gas inlet that extends through the nutrient media compartment to the sparging compartment to an inlet point near a central location of the flexible middle sheet to provide compressed gas to the sparging compartment; at least one nutrient media inlet port in fluid communication with the nutrient media compartment; and at least one compressed gas outlet port in fluid communication with the nutrient media compartment, wherein the porous flexible middle sheet allows the compressed gas from the gas inlet to aerate the nutrient media compartment from the sparging compartment, and wherein a plurality of pores of the flexible middle sheet are arranged so that pores located proximal to the inlet point near the central location of the flexible middle sheet are less densely dispersed than pores located proximal to an outer edge of the flexible middle sheet. PDF

US 9,469,671 | Closed Bioreactor.

 Single-use closed bioreactors with recirculating exhaust gas that can be operated in an uncontrolled environment are reported for the manufacturing of biological products using genetically modified biological cultures that produces carbon dioxide or that requires carbon dioxide in their metabolic process. A closed bioreactor, suitable for use in uncontrolled environment, comprising: a) a container comprising a nutrient medium and a biological culture; b) at least one gas outlet tube comprising an in line cartridge connected to the gas outlet comprising a carbon dioxide absorbent material; c) at least one gas inlet connected to the container comprising a tube for introducing gas into the container and wherein the gas inlet is also connected to the gas outlet through a tube allowing the gas to recirculate within the container, and wherein the gas inlet is connected to a source of gas; d) at least one liquid port for adding liquid to the container, wherein the liquid port is connected to the container and comprises a tube extending below the surface of the nutrient medium; e) a sterilizing filter capable of removing contaminants installed between the source of gas and the gas inlet; f) a stopcock installed between the sterilizing filter (e) and the gas inlet; g) a sterilizing filter capable of removing contaminants installed at the liquid port; h) a stopcock installed between sterilizing filter (g) and the liquid port; i) a peristaltic pump installed in the tube connected between the gas outlet and the gas inlet; and j) at least one sparger connected to the gas inlet and located in the container. PDF

US 9,469,426 | Single-Use Stationary Bioreactors and Mixing Vessels. 

Stationary bioreactors and mixing vessels fitted with single-use flexible bags utilizing wave hydrodynamic principle are described for use in every type of biological process and products. A method of inducing a wave-type mixing motion inside a flexible container comprising: a. supplying a flat, horizontal stationary surface; b. placing a flexible container comprising a top and bottom surface on the stationary surface, wherein the flexible container is capable of containing a liquid medium; c. pivotally attaching at least one movable flap to the stationary surface, wherein the movable flap comprises a rigid surface attached to the stationary surface by a hinge that allows the rigid surface to pivot to apply sufficient force to compress the top surface of the flexible container along an edge and then reverse direction of movement of the flap; d. operating the at least one movable flap periodically such that a wave type mixing motion of the liquid medium occurs within the flexible container; and e. wherein the at least one movable flap includes a first moveable flap and a second moveable flap, the first moveable flap mechanically connected to the second moveable flap such that movement of the first moveable flap causes movement of the second moveable flap. 2. A method of inducing a reciprocal mixing motion inside a flexible container comprising: a. supplying a flat, horizontal stationary surface; b. placing a flexible container comprising a top and bottom surface on the stationary surface, wherein the flexible container is capable of containing a liquid medium; c. pivotally attaching at least two movable flaps at opposite ends of the stationary surface, wherein the movable flaps each comprise a rigid surface attached to the stationary surface by a hinge that allows the rigid surface to pivot to apply sufficient force to compress the top surface of the flexible container along an edge and then reverse direction; d. operating the at least two movable flaps periodically such that a reciprocal mixing motion of the liquid medium occurs within the flexible container; and e. wherein the at least two movable flaps are mechanically connected such that movement of one of the at least two movable flaps causes movement of the other of the at least two movable flaps. 3. A method of inducing an orbital mixing motion inside a flexible container comprising: a. supplying a flat, horizontal stationary surface; b. placing a flexible container comprising a top and bottom surface on the stationary surface, wherein the flexible container is capable of containing a liquid medium; c. pivotally attaching four movable flaps to the stationary surface, wherein the flaps are attached to the stationary surface by hinges that allow the flaps to pivot to apply sufficient force to compress the top surface of the flexible container sequentially along each edge and then reverse direction; d. operating the four movable flaps periodically such that an orbital mixing motion of the liquid medium occurs within the flexible container; and e. wherein opposite flaps of the four movable flaps are mechanically connected such that movement of one of the opposite flaps causes movement of the other of the opposite flaps. 4. A method of inducing a vertical mixing motion inside a flexible container comprising: a. supplying a flat, horizontal stationary surface; b. placing a flexible container comprising a top and bottom surface on the stationary surface, wherein the flexible container is capable of containing a liquid medium; c. pivotally attaching two movable flaps at opposite ends of the stationary surface, wherein the flaps are attached to the stationary surface by hinges that allow the flaps to pivot to apply sufficient force to compress the top surface of the flexible container simultaneously along an edge and then reverse direction d. operating the two movable flaps periodically such that a vertical mixing motion of the liquid medium occurs within the flexible container; and e. wherein the two movable flaps are mechanically connected such that movement of one of the two movable flaps causes movement of the other of the two movable flaps. PDF

US 9,339,026 | Pneumatically Agitated and Aerated Single-Use Bioreactor. 

A single-use round flexible mixing bag for use in bioprocessing in which a fluid is received and agitated using an internal fluid-agitating element comprising a radial flow impeller driven by an internal pneumatic vane motor is disclosed. The bag may include an integral sparger and sensor receiver. Related methods are also disclosed. A disposable bioprocessing apparatus intended for receiving a fluid in need of agitation and sparging using a gas, comprising: a round, flexible bag having a round upper sheet with an interior and an exterior surface and an edge and a round lower sheet with an interior and an exterior surface and an edge and wherein the edge of the upper sheet and the edge of the lower sheet are sealed together to form a cavity capable of receiving and holding the fluid; a gas-driven fluid agitating element for agitating the fluid and assisting in distributing the bubbles comprising a pneumatic motor with a gas inlet and a gas outlet, a shaft and at least one rotating blade attached to the shaft, wherein said gas-driven fluid agitating element is located inside the flexible bag; a hard support surface affixed at the center of the interior surface of the lower sheet of the bag and to which the gas-driven fluid agitating element is fixed; a source of compressed gas fluidly connected to the gas inlet of the pneumatic motor, the source of compressed gas supplying compressed gas to the pneumatic motor to drive the at least one rotating blade to agitate the fluid; a sparger fluidly connected to the gas outlet of the pneumatic motor for forming bubbles, the sparger receiving exhaust gas from the pneumatic motor and aerating the fluid in the bag with the exhaust gas; a source of compressed gas to operate the pneumatic motor; a support surface to hold the bag; at least one liquid port; and at least one gas port. PDF

US 9,284,521 | Pivoting Pressurized Single-Use Bioreactor. 

Pressurized hermetically sealed bags disposed inside a cylindrical support and containing a septum with variable density of porosity and dividing the bag into two chambers are used to provide optimal mixing and gasification of nutrient media to grow a variety of biological cultures, particularly the cell cultures to produce a multitude of pharmaceutical and biotechnology products in a disposable system. A method for producing a biological product comprising: a) providing a bioreactor including: i) a cylindrical support with an inner surface configured to immovably retain a single-use flexible container with an inner volume and capable of holding nutrient media; ii) the single-use flexible container of (a)(i) having at least one interior wall, further including: a flexible septum with a surface immovably positioned within the flexible container and defining a right chamber and a left chamber, the septum having a plurality of pores with a variable density over the surface of the septum, wherein diameter of the pores range in size from 1 .mu.m to 1000 .mu.m, and said septum provides fluid communication between the right and the left chambers; iii) disposing the flexible container immovably inside the cylinder such that the septum is vertically positioned and a surface comprising 25-30% of the total surface of septum along the middle of the horizontal axis and the middle of the vertical axis has 3-5 times higher density of pores than the rest of the septum; iv) at least one liquid inlet; v) at least one liquid outlet; vi) at least one gas inlet in fluid communication with a source of compressed gas further comprising a sterilizing filter positioned between the source of compressed gas and the container; vii) at least one gas outlet capable of controlling the rate of flow of gas; viii) at least one sensor to measure the pressure inside the flexible container; ix) at least one sensor to measure the temperature of the liquids in the flexible container; x) a heater and/or a cooling element for heating and/or cooling the flexible container; xi) a motor driving a shaft connected to a rolling mechanism for pivoting the cylindrical support along its circular axis between -110 to +110 degrees; b) disposing the flexible container in the cylindrical support and immovably fixing the flexible container to the inner surface of the cylindrical support; c) introducing the nutrient media in the flexible container through the liquid inlet; d) introducing a biological culture capable of growing in the nutrient media and producing a biological product in the flexible container through the liquid inlet; e) heating or cooling the nutrient media to a pre-determined temperature; f) connecting the gas inlet to a source of a compressed gas; g) starting the flow of the compressed gas with sufficient pressure to achieve a pre-determined pressure in the flexible container that causes the flexible container to expand and stay in continuous contact with the inner surface of the cylindrical support; h) continuing the flow of compressed gas to maintain the pre-determined pressure in the flexible container; i) moving the shaft to pivot the cylindrical support between the angles from -110 and +110 degrees at a pre-determined frequency; j) monitoring the density and the viability of the biological culture in the nutrient media at predetermined time intervals; k) monitoring the concentration of the biological product in the nutrient media at predetermined time intervals; l) stopping the pivoting of the cylindrical support when the density of the biological culture or the concentration of the biological product reaches a pre-determined level and removing the nutrient media from the flexible container for further processing of the purification of the biological product in the nutrient media. PDF

US 9,238,789 | Baffled Single Use Bioreactor.

 A flexible disposable bioreactor having three, stagger-baffled compartments wherein the middle compartment houses a sparging rod is described to provide the highest degree of sparging and mixing to produce biological products. A disposable bioprocessing apparatus intended for receiving a fluid in need of agitation and sparging using a gas, comprising: a first sheet of film material having a first peripheral region; a second sheet of film material having a second peripheral region and overlapping said first sheet of film material such that said first and second peripheral regions of said first and second sheets of film material are joined together so as to form a peripheral region of a bag and to define an interior space within said bag between said first and second sheets of film material; at least two rows of staggered seals sealing said first sheet to said second sheet to create three baffled compartments, right, left and middle, within said bag, the at least two rows of staggered seals including a first row of spaced seals and a second row of spaced seals, the seals in the first row offset from the seals in the second row in a direction substantially perpendicular to a direction of at least one of the first row and the second row; at least one sparging rod disposed in said middle compartment and connected to a source of sterile compressed gas; a hard surface to support said bag; at least one fluid port in said bag; at least one gas exhaust port in said bag; and at least one gas inlet port in said bag. PDF

US 9,068,215 | Interconnected Bioreactors.

A method of homogeneously mixing the contents of a plurality of bioreactors by providing a receiving container capable of holding an appropriate quantity of the liquid and repeatedly raising and lowering the receiving container to a position above or below the position of the bioreactors resulting in mixing the contents of the bioreactors. A method of homogeneously mixing the contents of a plurality of bioreactors comprising: (a) providing a plurality of bioreactors, 1 through nth, in a series, installed on a same plane either in a series or in a circular pattern, the bioreactors comprising a bioreactor container with at least two surfaces and two opposing sides, to hold a liquid further comprising a nutrient media and a biological culture, a means of gassing the liquid further comprising a gas inlet, a gas sterilizing filter and a gas sparging filter, a gas exhaust, at least one nutrient media inlet/outlet, at least one liquid ports installed on the bottom with means of closing or opening the liquid port, a means of agitating the nutrient media, a plurality of sensors installed in the bioreactor container with at least a sensor for measuring the weight of the bioreactor container; (b) providing a receiving container capable of holding an appropriate quantity of the liquid wherein the receiving container further comprises a support capable of being raised or lowered; (c) connecting the liquid port of each bioreactor to the receiving container; (d) operating the bioreactors; (e) opening all liquid ports; (f) lowering the receiving container support to a position below the position of bioreactors; (g) measuring the increase in the weight of the receiving bioreactor to a pre-determined level; (h) raising the receiving container support to a position higher than the bioreactors; (i) measuring the decrease in the weight of the receiving container; (j) repeating the steps (f) to (i) periodically or continuously. PDF

US 8,668,886 | Separative Bioreactor. 

A bioreactor that combines the steps of recombinant expression and separation of a biological product by binding the secreted biological product with a resin, discarding the nutrient medium and eluting the biological product as a concentrated solution, eliminating the steps of sterile filtration and volume reduction. The method also allows loading of resin for column-purification, eliminating all steps of perfusion process and maintaining a sink condition of a toxic product in nutrient medium to optimize productivity of host cells. The instant invention also allows harvesting of solubilized inclusion bodies after the cells have been lysed and refolding of proteins inside the bioreactor. A separative bioreactor comprising: (a) a container having at least one interior wall; (b) a nutrient medium comprising a biological culture in the container; (c) at least one pouch comprising a porous material with a plurality of pores and a binding resin, wherein the pouch is placed inside the container; (d) at least one resin inlet/outlet directly connected to the pouch; (e) at least one liquid drain; (f) at least one gas inlet; (g) at least one gas outlet; (h) at least one aerator; (i) at least one agitator; (j) at least one heating or cooling element; (k) at least one temperature controller; and (l) at least one sensor. PDF

US 12/618,330 | Bioreactors for Fermentation and Related Methods.

 Bioreactors suitable for housing a predetermined volume of liquid comprising nutrient medium and biological culture comprising: (a) a container having at least one interior wall; (b) at least one nutrient medium inlet; (c) at least one liquid outlet; (d) at least one gas inlet; (e) at least one gas outlet; and (f) at least one cylindrical sparging filter attached to the at least one gas inlet, wherein the sparging filter comprises a plurality of pores along its axis which permit gas to be emitted radially from the sparging filter into the liquid, wherein the diameter of the plurality of pores does not exceed about 50 mu and wherein the orientation of the at least one sparging filter within the container provides for immersion of the plurality of pores within the liquid and substantially uniform distribution of emitted gas throughout the liquid, and related methods of using said bioreactors to prepare various biological products. PDF

US 15/404,899 Multipurpose Bioreactor. 

A multiuse bioreactor that is a single-use bioreactor, a development bioreactor, a commercial manufacturing bioreactor, a batch, a fed-batch, a perfusion and continuous bioreactor, a convective heat bioreactor, a product capture bioreactor, an ISO 9 bioreactor, a eukaryotic bioreactor, a prokaryotic bioreactor, a technology transfer-free bioreactor, and an inexpensive bioreactor is disclosed. A bioreactor comprising:  at least one single-use container with an inner volume, capable of holding a cell culture and culture medium to express a biological product, a top surface, and a bottom surface; at least one liquid inlet disposed in the top surface of the container in fluid communication with a plurality of sources of liquids including a culture medium, a cell culture, a pH adjusting solution, and a nutritive solution; at least one gas inlet disposed in the top surface of the container and in fluid communication with a source of at least one nutritive gas and at least one inert gas; an inline gas heater or cooler connected to the gas inlet; an inline gas sterilizing filter connected to the gas inlet; at least one gas sparging unit connected to the gas inlet and disposed in the culture medium; at least one exhaust gas outlet disposed in the top surface of the container and in fluid communication with outside environment further comprising a one-way exhaust gas flow control valve and an inline vent fan; a pressure sensor disposed in the container and connected to an electronic controller to adjust exhaust gas flow to maintain a positive pressure inside the container continuously; at least one liquid outlet disposed in the bottom surface of the container, further comprising a liquid flow control valve; a movable raised support platform to hold the container with an opening to allow passage of the liquid outlet to pass through the support platform; a mechanical device connected to the platform for shaking, rotating, rocking or vibrating the platform; a plurality of sensors in communication with an electronic controller to allow control of the condition of a liquid in the container; a capture column disposed under the support platform and above ground, holding a binding resin and connected to the liquid outlet to receive liquid from the container further comprising a process liquid inlet, a process liquid outlet and a process liquid outlet control valve. PDF

US 15/053,804 | Pivoting Pressure Single Use Bioreactor.

Pressurized hermetically sealed bags disposed inside a cylindrical support and containing a septum with variable density of porosity and dividing the bag into two chambers are used to provide optimal mixing and gasification of nutrient media to grow a variety of biological cultures, particularly the cell cultures to produce a multitude of pharmaceutical and biotechnology products in a disposable system. A mixing device comprising: a) a cylindrical support with an inner surface configured to immovably retain a single-use flexible container; b) a single-use flexible container capable of containing a liquid comprising: i) A flexible septum with an inner surface immovably positioned within the flexible container and defining a right chamber and a left chamber, the septum comprising a plurality of pores with a variable density over the surface of the septum; ii) At least one liquid inlet; iii) At least one liquid outlet; iv) At least one gas inlet in fluid communication with a source of compressed gas; v) At least one gas outlet capable of controlling the rate of flow of gas; vi) at least one sensor to measure the temperature of a liquid in the flexible container; and vii) A heater and/or a cooling element for controlling the temperature of the flexible container; c) a shaft for pivoting the cylindrical support along its circular axis between -110 to +110 degrees; and wherein, the flexible container is immovably fixed to the inner surface of the cylindrical support. PDF

US 14/249137 | Aeration Device for Bioreactors.

An aeration and mixing device for disposable flexible bioreactors comprising a mesh of interconnected perforated disposable tubes to form a structure to cover essentially the entire bottom surface of a disposable flexible bioreactor and wherein a continuous flow of gases through the perforations in the tubes provides an aeration and a mixing function. An aeration and mixing device for bioreactors comprising a plurality of perforated tubes interconnected to form a horizontal mesh disposed in a bioreactor, wherein the mesh is connected to one or more gas inlets. A disposable bioreactor comprising an aeration and mixing device comprising a plurality of disposable perforated tubes interconnected to form a horizontal mesh, wherein the mesh is connected to one or more gas inlets. PENDING ISSUANCE PDF

 

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Downstream Processing

Methods of capturing and purification of chemicals and drugs are refined to allow low cost manufacturing of biological products. Bioreactors are used for expressing a variety of chemicals and drugs; the next phase after upstream operation is capture and purification, the two steps that have historically required centrifugation and filtration, both of which cause significant loss of product and deterioration of product quality. The inventions described below use a method of first capturing the expressed product and then discarding the contents of bioreactor, eliminating two major costly steps. It is now possible to use a bioreactor to express and purify a product within the same container.

US 9,321,805 | Downstream Bioprocessing Device.

 Large-scale downstream processing of secreted recombinant proteins is provided in a single device, wherein the contents of a plurality of bioreactors are combined simultaneous to their harvesting and purification resulting in significant savings of time and the cost of manufacturing. A method for pooling, harvesting and purifying a target protein expressed by a biological culture in a nutrient media in a plurality of bioreactors comprising: a. Providing a downstream processing device comprising a cylinder capable of holding a liquid wherein the cylinder comprises: i. a top opening comprising a filter covering the 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 port valve and a top sampling port filter capable of removing a biological culture wherein said top sampling port is connected to one of the top liquid ports of (iii); iii. a bottom opening comprising a 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, and a plenum having a plurality of liquid ports wherein said ports are capable of closing and opening; iv. a bottom sampling port comprising a bottom sampling port valve and a bottom sampling port filter capable of removing a biological culture wherein said bottom sampling port is connected to the bottom liquid port of (e); v. a means of mixing the contents of the cylinder; and vi. sensors located at one or more places on or in the cylinder capable of detecting turbidity of the contents of the cylinder; b. setting the downstream processing device at such height that the top liquid port is below the lowest level of one or more bioreactors; c. connecting the liquid ports of the plenum of (iv) above to one or more bioreactors containing a nutrient media comprising a biological culture capable of producing a target protein; d. removing the top cap and adding to the cylinder a quantity of a chromatography media sufficient to bind substantially all of the target protein present in the nutrient media; e. replacing the top cap and opening the top and bottom flow control valves; f. filling the cylinder with the nutrient media under gravity pressure from the one or more bioreactors; g. closing the bottom flow control valve; h. mixing the chromatography media to achieve a uniform distribution in the cylinder as indicated by the measurement of turbidity by the sensors of (vi) above and adjusting the speed of mixing to achieve a uniform turbidity; i. 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 between the top and bottom sampling ports reaches about 1:100 and closing the bottom flow control valve when the ratio reaches about 2:100; j. maintaining the flow of nutrient media from the one or more bioreactors into the cylinder and allowing the nutrient media to flow out of the top liquid port until the contents of the bioreactors have passed through the cylinder; k. closing the bottom flow control valve and the liquid ports of the plenum and disconnecting the bioreactors from the plenum; l. opening the bottom flow control valve to allow the nutrient media in the cylinder to drain out through one of the unoccupied ports in the plenum and then closing that port; m. connecting one or more liquid ports of the plenum to at least one source of washing buffer suitable for removing debris in the cylinder; n. starting the flow of the washing buffer into the cylinder through the plenum and allowing the washing buffer to drain out of the top liquid port until a desired level of debris is reached in the drained wash buffer; o. stopping the flow of the washing buffer; p. opening one of the unoccupied liquid ports in the plenum to allow the washing buffer to drain out of the cylinder and closing that liquid port of the plenum; q. connecting one or more liquid ports of the plenum to at least one source of elution buffer capable of breaking the binding of the target protein to the chromatography media; r. filling the cylinder with elution buffer and then stopping the flow of elution buffer into the plenum; s. closing the bottom flow control valve and begin mixing; t. 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; u. stopping the mixing; v. opening the bottom flow control valve and one of the unoccupied liquid ports in the plenum and collecting the elution buffer in a container as a purified concentrated solution of the target protein. PDF

US 9,290,732 | Buoyant Protein Harvesting Device.

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. A protein-harvesting device comprising: a bioreactor including nutrient media containing a genetically modified organism or cells capable of producing a target protein for harvesting, the bioreactor configured to contain a fixed volume of the nutrient media and adjust at least one property of the fixed volume of nutrient media; a first container disposed in the bioreactor, the first container including a porous mesh enclosing a substantially hollow interior compartment, wherein the mesh comprises a plurality of pores allowing exchange of liquid with the hollow interior compartment, and the hollow interior compartment comprises a suitable amount of a chromatography media capable of binding the target protein; and a buoyancy device attached to the first container to allow the first container to float within the nutrient media contained in the bioreactor. PDF

US 9,284,346 | Preparative Chromatography Column and Methods.

 A chromatography column that captures components in a process liquid in a free flow state and allows elution in steps is described. A disposable preparative chromatography column for effecting separation of components of a process liquid comprising: a. a flexible disposable bag with an inner volume; b. a septum disposed within the flexible disposable bag to separate the flexible disposable bag into a flexible upper chamber and a flexible lower chamber, the septum including a porous surface that allows passage of liquid but not chromatography media from the upper chamber to the lower chamber; c. a chromatography media disposed in the upper chamber; d. at least one inlet to introduce liquid into the flexible upper chamber of the bag containing the chromatography media; e. at least one outlet to remove the liquid from the flexible lower chamber of the bag after passage through the septum; and f. at least one mixing mechanism to mix the contents of the bag. PDF

US 9,200,335 | Separative Bioreactor.

 A bioreactor that combines the steps of recombinant expression and separation of a biological product by binding the secreted biological product with a resin, discarding the nutrient medium and eluting the biological product as a concentrated solution, eliminating the steps of sterile filtration and volume reduction. The method also allows loading of resin for column-purification, eliminating all steps of perfusion process and maintaining a sink condition of a toxic product in nutrient medium to optimize productivity of host cells. The instant invention also allows harvesting of solubilized inclusion bodies after the cells have been lysed and refolding of proteins inside the bioreactor. 1. A method for producing and separating a biological product from a liquid comprising: a. providing a separative bioreactor comprising: (1) a container suitable for growing a biological culture, wherein the container further comprises a liquid inlet and a drain; and (2) at least one pouch comprising a porous material with a plurality of pores and capable of housing a binding resin, wherein the pouch is placed inside the container, and wherein the pouch optionally comprises at least one resin inlet/outlet port directly connected to the pouch; b. introducing nutrient medium into the container; c. introducing biological culture into the container; d. growing of the biological culture by agitating the liquid; e. determining the concentration of biological product in the liquid at periodic intervals; f. introducing resin into the pouch, when the concentration of biological product reaches a desired amount, through the resin inlet sufficient to bind the biological product determined in step (e); g. determining the concentration of biological product in the liquid until it reaches a predetermined low value; h. removing the liquid from the container through the drain and discarding it; and (1) adding to the container a sufficient quantity of a buffer capable of removing the biological product from the resin and collecting the buffer for further purification of the biological product; or (2) removing the resin by aspirating the resin through the resin port and subjecting the resin to further steps of purification outside the container. 2. A method for producing and separating a biological product from a volume of liquid comprising: a. providing a Separative bioreactor comprising: (1) a container suitable for growing a biological culture, wherein the container further comprises a liquid inlet and a drain; and (2) at least one pouch comprising a porous material with a plurality of pores and capable of housing a binding resin, wherein the pouch is placed inside the container, and wherein the pouch optionally comprises at least one resin inlet/outlet port directly connected to the pouch; b. introducing nutrient medium into the container; c. introducing biological culture into the container; d. growing of the biological culture by agitating the liquid; e. determining the concentration of biological product in the liquid at periodic intervals; f. introducing resin into the pouch at periodic intervals during growth of the biological culture through the resin inlet sufficient to bind the biological product determined in step (e); g. continue growth of the biological culture; h. continue binding of the biological product to the resin until the concentration of biological product in the liquid reaches a desired low value; i. removing the liquid from the container through the drain and discarding it; and (1) adding to the container a sufficient quantity of a buffer capable of removing the biological product from the resin and collecting the buffer for further purification; or (2) removing the resin by aspirating the resin through the resin port and subjecting the resin to further steps of purification outside the container. 3. A method for producing and separating a biological product from a volume of liquid comprising: a. providing a Separative bioreactor comprising: (1) a container suitable for growing a biological culture, wherein the container further comprises a liquid inlet and a drain; and (2) at least one pouch comprising a porous material with a plurality of pores and capable of housing a binding resin, wherein the pouch is placed inside the container, and wherein the pouch optionally comprises at least one resin inlet/outlet port directly connected to the pouch; b. introducing nutrient medium into the container; c. introducing biological culture into the container; d. growing of the biological culture by agitating the liquid; e. determining the density of the biological culture in the liquid at periodic intervals; f. when the density of the biological culture reaches a predetermined high value, adding to the liquid a chemical solution capable of lysing the cells to release inclusion bodies into the liquid; g. adding to the liquid, a chemical solution capable of solubilizing the inclusion bodies; h. adding sufficient quantity of a resin to the pouch to bind substantially all of the solubilized inclusion bodies; i. removing the liquid from the container through the drain and discarding it; and (1) adding sufficient quantity of a chemical solution in the container to break the bond between the solubilized inclusion bodies and the resin and collecting the chemical solution containing solubilized inclusion bodies through the drain for further purification; or (2) removing the resin by aspirating the resin through the resin port. PDF

US 8,932,843 | Buoyant Protein Harvesting Device.

 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. 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. PDF.

US 8,852,435 | Purification and Separation Treatment Assembly (PASTA) For Biological Products. 

An assembly capable of capturing and purifying expressed biological products during or at the end of a bioreaction cycle is disclosed wherein a binding resin is kept separated from the contents of the bioreactor allowing capturing, harvesting and purification of biological products in a bioreactor; the invention additionally provides means of removing undesirable metabolic products as well as provides for efficient loading of chromatography columns. A method for harvesting and purifying a target biological product at the end of a production cycle comprising: a) Expressing a target biological product in a bioreactor; b) Disposing in the bioreactor at the end of the production cycle, an appropriate length of a purification and separation treatment assembly (PASTA) comprising: a flexible porous tube filled with a binding resin, wherein the porous tube comprises a plurality of pores smaller in size than the size of the binding resin and wherein the porous tube is sealed at both ends to provide a pre-determined quantity of resin; c) Capturing the target biological product by mixing the contents of the bioreactor to allow substantially complete binding of the target biological product to the resin in the PASTA; d) Removing the contents of the bioreactor except the PASTA from the bioreactor; e) Eluting the target biological product from the PASTA held in the bioreactor by adding to the bioreactor a buffer capable of removing the binding of the target biological product from the resin in the PASTA. PDF

US 8,668,886 | Separative Bioreactor.

 A bioreactor that combines the steps of recombinant expression and separation of a biological product by binding the secreted biological product with a resin, discarding the nutrient medium and eluting the biological product as a concentrated solution, eliminating the steps of sterile filtration and volume reduction. The method also allows loading of resin for column-purification, eliminating all steps of perfusion process and maintaining a sink condition of a toxic product in nutrient medium to optimize productivity of host cells. The instant invention also allows harvesting of solubilized inclusion bodies after the cells have been lysed and refolding of proteins inside the bioreactor. PDF

US 8,506,797 | Downstream Bioprocessing Device.

 Large-scale downstream processing of secreted recombinant proteins is provided in a single device, wherein the contents of a plurality of bioreactors are combined simultaneous to their harvesting and purification resulting in significant savings of time and the cost of manufacturing. A downstream processing device for pooling, harvesting and purifying a target protein comprising: a. a cylinder with inner volume and having a top opening, a bottom opening and suitable for holding a nutrient media, a biological culture and a chromatography media; b. a means for retaining the chromatography media in the cylinder comprising a top filter covering the top opening and a bottom filter covering the bottom opening of the cylinder; c. a removable top cap to hold the top filter in place comprising at least one top liquid port and a top flow control valve; d. a top sampling port comprising a top sampling port valve and a top sampling port filter capable of removing the biological culture wherein said top sampling port is connected to one of the top liquid ports of (c); e. a removable bottom cap to hold the bottom filter disk in place comprising a bottom liquid port, a bottom flow control valve, and a plenum having a plurality of liquid ports with means for closing and opening the plurality of liquid ports; f. a bottom sampling port comprising a bottom sampling port valve and a bottom sampling port filter to capable of removing the biological culture wherein said bottom sampling port is connected to the bottom liquid port of (e); g. a mixing means for keeping the chromatography media continuously and uniformly suspended throughout the cylinder; h. a means for continuously measuring the turbidity of the contents of the cylinder in a plurality of locations in the cylinder; i. a means for continuously measuring the ratio of the concentration of a target protein produced by the biological culture in the nutrient media exiting the top liquid port as compared to the concentration of the target protein entering the bottom liquid port; j. a means for supporting the cylinder vertically; k. a means for heating or cooling the contents of the cylinder; l. a temperature sensor to measure the temperature of the nutrient media in the cylinder; m. an electronic means for automatically controlling the opening and closing of the bottom flow control valve in response to the ratio of the concentration of the target protein in the top and bottom liquid ports, the means for heating and cooling the contents of the cylinder in response to the temperature of the nutrient media and controlling the intensity of the mixing means in response to the differences in the measurement of turbidity in the cylinder.  PDF

US 8,183,035 | Single Container Manufacturing of Biological Product.

 A method of manufacturing biological products in a single container from the growth of cells to purification of the product is performed in a flexible disposable bioreactor that uses only a compressed gas for mixing and gasification. A porous septum is used to create the gasification and mixing as well as to separate a chromatography media used to harvest and purify a biological product in the same container. The closed container can be used in any environment without the risk of contamination to the product or the risk of contamination of environment with the product. This allows large manufacturing of hazardous substances, drugs and vaccines anywhere at the lowest cost possible. Numerous applications can be found in counter-terrorism and biodefense operations as well in managing epidemic illnesses. A method for producing and purifying a biological product in an uncontrolled environment comprising: providing a disposable container for housing a nutrient media, the disposable container comprising an upper chamber and a lower chamber separated by a porous flexible membrane with a plurality of pores ranging from 10 microns to 1000 microns, at least one gas inlet port in the lower chamber, at least one liquid outlet port in the lower chamber, at least one gas exhaust port in the upper chamber, at least one liquid inlet port in the upper chamber, a structure for supporting the disposable container, one or more sensors for sensing one or more parameters of the nutrient media in the disposable container, and a heater for heating the contents of the disposable container, the heater having a thermostat; adding to the disposable container the nutrient media and a biological culture capable of secreting the biological product, through the liquid inlet in the upper chamber; starting and continuing supplying a gas through the gas inlet in the lower chamber of the disposable container to mix the nutrient media and the biological culture and to achieve and maintain a pre-determined level of gas concentration in the nutrient media; adding a sufficient quantity of a chromatography media having a particle size larger than the diameter of the pores in the porous flexible membrane through the liquid inlet in the upper chamber when a pre-determined level of the biological product has reached in the nutrient media to bind essentially all of the biological product in the nutrient media; removing and discarding the nutrient medium and the biological culture through the liquid outlet in the lower chamber; adding at least once a buffer suitable for disassociating the biological product from the chromatography media in the disposable container through the liquid inlet in the upper chamber; removing the buffer through the liquid outlet as a concentrated purified solution of the biological product. PDF

US 15/135,263 | Downstream Bioprocessing Device.

 Large-scale downstream processing of secreted recombinant proteins is provided in a single device, wherein the contents of a plurality of bioreactors are combined simultaneous to their harvesting and purification resulting in significant savings of time and the cost of manufacturing. 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. PDF

US 15/076,337 | Buoyant Protein Harvesting Device.

 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 for harvesting refolded protein comprising: a. providing a 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 protein in a refolding solution; c. adjusting the buoyancy of the protein-harvesting device to a pre-determined level; d. capturing a refolded protein from the refolding solution with the protein harvesting device; e. removing the refolding solution; f. eluting the refolded protein from the chromatography media of the protein harvesting device; and g. reconstituting the refolded protein eluted from the chromatography media for further purification. 2. A method for continuous harvesting proteins comprising: a. providing a 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 bioreactor comprising a genetically modified organism or cells capable of producing a protein for harvesting in a nutrient media at the beginning of protein production; c. adjusting the pH and electrolyte concentration of the nutrient media to optimize the binding of the protein to the chromatography media; d. capturing the protein from the nutrient media continuously throughout; and e. removing the protein-harvesting device from the nutrient media for further processing of the protein bound to the chromatography media. 3. A method for continuous harvesting proteins comprising: a. providing a 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 bioreactor comprising a genetically modified organism or cells capable of producing a protein for harvesting in a nutrient media at the beginning of protein production; c. adjusting the pH and electrolyte concentration of the nutrient media to optimize the binding of the protein to the chromatography media; d. capturing the protein from the nutrient media continuously throughout production of the target protein; and e. removing the protein-harvesting device from the nutrient media for further processing of the protein bound to the chromatography media. 4. A method comprising: a. continuously removing expressed protein from a nutrient media to enhance the level of expression that may be depressed because of the higher concentration of protein in the mixture; b. providing a 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; c. disposing the protein-harvesting device in a bioreactor comprising a genetically modified organism or cells capable of producing a protein for harvesting in a nutrient media at the beginning of protein production; d. adjusting the pH and electrolyte concentration of the nutrient media to optimize the binding of the protein to the chromatography media; e. capturing the protein from the nutrient media continuously throughout; and f. removing the protein-harvesting device from the nutrient media for further processing of the protein bound to the chromatography media. 5. A method comprising: a. continuously removing expressed protein from a nutrient media to reduce the toxicity of the expressed protein to host cells and thus prolonging the cycles of expression substantially increasing the yields of production; b. providing a 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; c. disposing the protein-harvesting device in a bioreactor comprising a genetically modified organism or cells capable of producing a protein for harvesting in a nutrient media at the beginning of protein production; d. adjusting the pH and electrolyte concentration of the nutrient media to optimize the binding of the protein to the chromatography media; e. capturing the protein from the nutrient media continuously throughout; and f. removing the protein-harvesting device from the nutrient media for further processing of the protein bound to the chromatography media. 6. A method comprising: a. increasing the chemical stability of expressed protein by binding it to a chromatography media as soon as it is expressed as the chemicals are always less stable in a solution form than in a solid form or in this case a complex form; b. providing a 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; c. disposing the protein-harvesting device in a bioreactor comprising a genetically modified organism or cells capable of producing a protein for harvesting in a nutrient media at the beginning of protein production; d. adjusting the pH and electrolyte concentration of the nutrient media to optimize the binding of the protein to the chromatography media; e. capturing the protein from the nutrient media continuously throughout; and f. removing the protein-harvesting device from the nutrient media for further processing of the protein bound to the chromatography media. PDF

US 15/050,982 | Preparative Chromatography Column and Methods.

 A chromatography column that captures components in a process liquid in a free flow state and allows elution in steps is described. A method of chromatographic separation of a desired component comprising: a. Providing a disposable preparative chromatography apparatus comprising: i. a flexible disposable bag with an inner volume; ii. a septum disposed within the flexible disposable bag to separate the flexible disposable bag into a flexible upper chamber and a flexible lower chamber, the septum including a porous surface that allows passage of liquid but not chromatography media from the upper chamber to the lower chamber; iii. a chromatography media disposed in the upper chamber; iv. at least one inlet to introduce liquid into the flexible upper chamber of the bag containing the chromatography media; v. at least one outlet to remove the liquid from the flexible lower chamber of the bag after passage through the septum; and vi. at least one mixing mechanism to mix the contents of the bag; b. Introducing a quantity of liquid containing a component to be separated into the upper chamber of the flexible bag through the inlet of (iv) above; c. Mixing the contents of the flexible bag to allow the component to bind to the chromatography media; d. Opening the drain in the lower chamber and draining out the liquid that has passed through the septum; e. Closing the drain; f. Introducing into the upper chamber a solution capable of disrupting the binding between the component and the chromatography media; g. Mixing the contents of the column until the component has been sufficiently released from the chromatography media; h. Opening the drain in the lower chamber; i. Collecting the contents of the flexible bag as a concentrated solution of the component. PDF

US 15/050,982 | Preparative Chromatography Column and Methods.

A chromatography column that captures components in a process liquid in a free flow state and allows elution in steps is described. A method of chromatographic separation of a desired component comprising: a. Providing a disposable preparative chromatography apparatus comprising: i. a flexible disposable bag with an inner volume; ii. a septum disposed within the flexible disposable bag to separate the flexible disposable bag into a flexible upper chamber and a flexible lower chamber, the septum including a porous surface that allows passage of liquid but not chromatography media from the upper chamber to the lower chamber; iii. a chromatography media disposed in the upper chamber; iv. at least one inlet to introduce liquid into the flexible upper chamber of the bag containing the chromatography media; v. at least one outlet to remove the liquid from the flexible lower chamber of the bag after passage through the septum; and vi. at least one mixing mechanism to mix the contents of the bag; b. Introducing a quantity of liquid containing a component to be separated into the upper chamber of the flexible bag through the inlet of (iv) above; c. Mixing the contents of the flexible bag to allow the component to bind to the chromatography media; d. Opening the drain in the lower chamber and draining out the liquid that has passed through the septum; e. Closing the drain; f. Introducing into the upper chamber a solution capable of disrupting the binding between the component and the chromatography media; g. Mixing the contents of the column until the component has been sufficiently released from the chromatography media; h. Opening the drain in the lower chamber; i. Collecting the contents of the flexible bag as a concentrated solution of the component. PDF

US 14/569, 012 | Buoyant Protein Harvesting Device.

 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. A protein-harvesting device comprising: a. A first container comprised of a porous mesh enclosing a substantially hollow interior compartment, wherein the mesh comprises a plurality of pores allowing exchange of liquid with the interior compartment; b. The hollow interior compartment comprises a suitable amount of a chromatography media capable of binding a target protein; and c. A buoyancy device attached to the porous mesh container; wherein the protein harvesting device is further disposed inside a second container comprising nutrient media containing a genetically modified organism or cells capable of producing the target protein for harvesting. PDF

US 14/491, 651 | Harvesting and Purification or Perfusion Yielder (HAPPY) Device.

 A modular device comprising one or more porous substrate subunits comprising a binding substrate that can interact with a target biological product, either during, or at the end of a manufacturing cycle; and methods of using the device to harvest or purify a biological product. A harvesting and purification or perfusion yielding (HAPPY) device comprising: a top housing subunit and a bottom housing subunit adapted to connect to each other, each comprising a port to facilitate the flow of liquid into and out of the device; at least one substrate subunit having a top and bottom surface, disposed between the top and bottom subunits, wherein the interior cavity of the substrate subunit is defined by a housing wall and wherein the top and bottom subunits comprise a porous mesh material covering; and the interior cavity of the substrate subunit comprises a volume of binding substrate capable of binding a target product. PDF

US 13/754,167 | Separative Harvesting Device.

 A harvesting device for capturing a biological product directly by binding the secreted biological product with a resin, discarding the nutrient medium and eluting the biological product as a concentrated solution, eliminating the steps of sterile filtration and volume reduction, thus allowing one to combine the steps of recombinant expression and separation of a biological product. The method allows loading of resin for column-purification, eliminating all steps of perfusion process and maintaining a sink condition of a toxic product in nutrient medium to optimize productivity of host cells. The instant invention also allows harvesting of solubilized inclusion bodies after the cells have been lysed and refolding of proteins inside the bioreactor. A harvesting device for capturing a biological product comprising at least one container with at least one external surface and an inner volume to hold at least one ligand or resin capable of binding a biological product and the surface having a plurality of pores. PDF

US 13/246830 | Preparative Chromatography Column and Methods.

A chromatography column that captures components in a process liquid in a free flow state and allows elution in steps is described. A preparative chromatography column for effecting separation of components of a liquid comprising: a. a housing with an inner volume; b. a chromatographic media disposed inside the housing; c. at least one inlet to introduce the liquid into the housing; d. an outlet to remove the liquid from the housing; e. at least one filter attached to the outlet to retain the chromatography media from the housing; f. a means of mixing the contents of the housing. PDF

US 13/092,955 | Separative Bioreactor.

 A bioreactor that combines the steps of recombinant expression and separation of a biological product by binding the secreted biological product with a resin, discarding the nutrient medium and eluting the biological product as a concentrated solution, eliminating the steps of sterile filtration and volume reduction. The method also allows loading of resin for column-purification, eliminating all steps of perfusion process and maintaining a sink condition of a toxic product in nutrient medium to optimize productivity of host cells. The instant invention also allows harvesting of solubilized inclusion bodies after the cells have been lysed and refolding of proteins inside the bioreactor. A separative bioreactor suitable for housing a predetermined volume of liquid comprising nutrient medium and biological culture comprising: (a) a container having at least one interior wall; (b) at least one pouch made of a porous material with plurality of pores and said pouch having sufficient inner volume to house a predetermined quantity of a binding resin and said pouch placed inside said container; (c) at least one resin inlet/outlet directly connected to said pouch; (d) at least one nutrient medium inlet; (e) at least one liquid drain; (f) at least one gas inlet; (h) at least one gas outlet; and (i) at least one sparging filter; (g) at least one means of agitating the liquid; (h) at least one means of heating or cooling the liquid (i) at least one means of controlling the temperature of the liquid; (j) at least one sensor. PDF

US 13/083,589 | Protein Harvesting.

Methods of harvesting proteins directly from bioreactors to avoid at several steps in the purification of recombinant drugs are disclosed. A method of harvesting a target protein from a liquid in a first container comprising: A means of contacting said target protein with a resin capable of binding substantially all of said target protein to form a protein-resin conjugate; A means of separating said protein-resin conjugate from said liquid; A means of recovering said target protein from said protein-resin conjugate. PDF

14/507, 469 | Purification and Separation Treatment Assembly (PASTA) For Biological Products.

 An assembly capable of capturing and purifying expressed biological products during or at the end of a bioreaction cycle is disclosed wherein a binding resin is kept separated from the contents of the bioreactor allowing capturing, harvesting and purification of biological products in a bioreactor; the invention additionally provides means of removing undesirable metabolic products as well as provides for efficient loading of chromatography columns. A purification and separation treatment assembly (PASTA) for biological products expressed in a bioreactor comprising a flexible porous tube filled with a binding resin and having a plurality of pores smaller in size than the size of the binding resin and sealed at both ends and at regular distances along the length of the mesh tube. A method for harvesting and purifying a target biological product at the end of a production cycle comprising: a) Expressing the target biological product in a bioreactor; b) Disposing in the bioreactor at the end of the bioreaction cycle, an appropriate length of PASTA according to claim 1 to provide a pre-determined quantity of resin; c) Capturing the target biological product by mixing the contents of the bioreactor to allow substantially complete binding of the target biological product to the resin in the PASTA; d) Removing the contents of the bioreactor except the PASTA from the bioreactor; e) Eluting the target biological product from the PASTA held in the bioreactor by adding to the bioreactor, a buffer capable of removing the binding of the target biological product from the resin in the PASTA. A method for removing undesirable metabolites in a bioreaction cycle comprising: a) Adding to a bioreactor a sufficient quantity of a PASTA according to claim 1, containing a resin or a mixture of resins capable of binding the undesirable metabolic products in the bioreactor; b) Removing the PASTA from the bioreactor upon achieving a desired low level of undesirable metabolic components in the bioreactor; c) Reacting the removed PASTA with a buffer capable of removing undesirable metabolites and re-using the PASTA in step (a). A method for loading a chromatography column with a resin comprising: a) Winding to fill a sufficient quantity of the PASTA according to claim 1 in a reel of a height and a width of side plates are such that they fit in a chromatography column; b) Inserting the reel in a chromatography column. PDF

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Monitoring

Bioreactor monitoring using direct contact sensors introduced contamination risk; monitoring through remote signals with sensors embedded in the bioreactor products highly variable signals because of the changes in the properties of the culture medium. The invention described here solves the problem by disposing a cuvette inside a bioreactor that is accessible from outside allowing use of high sophisticated sensors that are re-usable and allow close monitoring of the conditions of bioreactor.

US 9,469,834 | Non-Invasive Bioreactor Monitoring.

 A pair or receptacles capable of housing an emitter probe and a detector probe installed inside a bioreactor to monitor the properties of the nutrient media without contacting the nutrient media.  A device for non-invasive, non-contact monitoring of a nutrient media in a bioreactor comprised of a container with a top surface, a bottom surface, and an inner volume capable of holding the nutrient media, the device comprising: at least one pair of receptacles suitable for housing probes, the receptacles configured for hermetically sealing the probes from the nutrient media; and each comprising one open end in fluid communication with the outside of the bioreactor via the bottom surface and one sealed end; an emitter probe capable of producing an electromagnetic or acoustic signal disposed in a first receptacle of the at least one pair of receptacles; a detector probe capable of detecting an electromagnetic or acoustic signal disposed in a second receptacle of the at least one pair of receptacles; wherein the distance between the first receptacle and the second receptacle can be varied by adjusting the first receptacle and the second receptacle up and down, or sideways, wherein the distance between the receptacles represents the depth of the nutrient media monitored; and an electronic means for converting a signal received from the detector probe into physical and chemical properties of the nutrient media. PDF

US 8,545,759 | Noninvasive Bioreactor Monitoring.

A pair or receptacles capable of housing an emitter probe and a detector probe are installed inside a bioreactor to monitor the properties of the nutrient media without contacting the nutrient media.  method for monitoring a nutrient media in a bioreactor comprising: a. Disposing in a bioreactor at least one pair of receptacles suitable for housing an emitter probe or a detector probe wherein each receptacle comprises one open end in fluid communication with the outside of the bioreactor and one sealed end; b. Disposing an emitter probe in a one receptacle of the pair of receptacles; c. Disposing a detector probe in the other receptacle of the pair of receptacles; d. Adjusting the direction of the emitter and the detector probes to face each other to read transmitted radiation or to an angle to read diffraction of radiation; e. Activating the emitter and detector probes to record transmission or diffraction of the radiation applied to the nutrient media of the bioreactor. PDF