Continuous processing methods for biological products

Abstract

The present invention is directed to the development of continuous processing technology for the purification of biopharmaceuticals and biological products, such as monoclonal antibodies, protein therapeutics, and vaccines. Methods for continuous processing of a biological product in a feed stream toward formulation of a purified bulk product are described.

What is claimed is: 
     
         1 . A process for production of a biological product comprising:
 (a) continuously transferring a biological product-containing solution to a first unit operation capable of separating the solution into two or more fractions comprising at least one fraction containing said biological product and at least one fraction containing one or more components of the biological product-containing solution desired to be separated from said biological product, wherein said first unit operation has at least one inlet and at least one outlet and is configured to permit continuous fluid flow between said at least one inlet and at least one outlet;   (b) continuously transferring the flow from at least one outlet of said first unit operation to a second unit operation having at least one inlet for receiving flow from at least one outlet of said first unit operation, said flow from said first unit operation comprising said biological product, said second unit operation capable of separating the product received through said at least one inlet into two or more fractions comprising at least one fraction containing said biological product and at least one fraction containing one or more components of the product received through said at least one inlet desired to be removed from said biological product,
 wherein said second unit operation further has at least one outlet and is configured to permit continuous flow between said at least one inlet and at least one outlet, and wherein the throughput of said second unit operation is equal to the output flow received from said first unit operation; 
   (c) recovering a solution containing said biological product.   
     
     
         2 . The process according to  claim 1 , wherein, independently for each transferring step, the transferring step comprises directing the flow from the preceding unit operation to an intermediate storage vessel, wherein said vessel has a holding capacity equal to or less than half of the volume received from the preceding unit operation during processing of one entire lot. 
     
     
         3 . A method for the manufacture of a biological product in a continuous process system comprising the steps:
 (a) producing conditioned media containing a biological product in a bioreactor;   (b) continuously removing cells and cell debris from said conditioned media to produce a clarified solution comprising said biological product;   (c) continuously transferring said clarified solution to a first unit operation comprising a capture chromatography process comprising (1) contacting said product solution with an affinity ligand capable of complexing with said biological product and (2) dissociating complexes of said affinity ligand and said biological product formed in said contacting step to produce a purified biological product solution;   (d) continuously transferring the purified biological product solution to a second unit operation comprising a low pH incubation process comprising (1) collecting at least part of said purified biological product solution in an incubation receptacle, (2) adjusting the pH of said purified biological product solution to a pH calculated to inactivate any viruses contained in said solution, and (3) readjusting the pH of said purified biological product solution to produce a virus-inactivated biological product solution;   (e) continuously transferring said virus-inactivated biological product solution to at least one addition unit operation comprising a process selected from anion exchange chromatography, cation exchange chromatography, nanofiltration, ultrafiltration/difiltration, microfiltration, or combinations thereof in any order; and   (f) recovering a solution containing said biological product.   
     
     
         4 . The method according to  claim 3 , wherein said removal of cells and cell debris in step (b) comprises centrifugation. 
     
     
         5 . The method according to  claim 3 , wherein said first unit operation is a simulated moving bed affinity chromatography process. 
     
     
         6 . A method for incubating a solution for a predetermined period of time at desired solution conditions, comprising:
 (a) continuously transferring a biological product solution into a first incubation receptacle;   (b) adjusting the solution conditions to desired incubation conditions in said first receptacle;   (c) continuously transferring the biological product solution from said first receptacle into at least one holding receptacle and holding the solution in said at least one holding receptacle;   (d) continuously transferring the biological product solution from a holding receptacle into a readjustment receptacle;   (e) adjusting the solution conditions in said readjustment receptacle to desired post-incubation conditions for the biological product solution;   wherein the residence time of the biological product solution in said at least one holding receptacle, plus the transfer time from said first receptacle to said at least one holding receptacle, plus the transfer time from said at least one holding receptacle to said readjustment receptacle is controlled to match said predetermined period of time, and wherein said incubation conditions are different from said post-incubation conditions.   
     
     
         7 . A method for incubating a biological product solution at desired solution conditions for a controlled period of time and within a desired tolerance time window, comprising:
 (a) continuously transferring a biological product solution into a series of incubation receptacles such that (the volume of each receptacle) (the flow rate)×(the desired tolerance time),   (b) after filling of each individual receptacle, discontinuing transfer to that receptacle and adjusting the solution conditions to the desired incubation condition in said receptacle,   (c) after achieving the desired incubation condition in step (b), holding that receptacle for the controlled period of incubation,   (d) after said period of incubation in step (c), readjusting the solution conditions in that receptacle to a desired post-incubation condition different from said incubation condition, and   (e) after achieving the desired post-incubation condition, continuously transferring the biological product solution out of said series of incubation receptacles.   
     
     
         8 . The method according to  claim 6  or  claim 7 , wherein one or more of said receptacles further comprises mixing means for ensuring a homogenous solution within said receptacle. 
     
     
         9 . The method according to  claim 8 , wherein said mixing means is a rocking table. 
     
     
         10 . The method of  claim 6  or  claim 7 , wherein one or more of said receptacles further comprises a vent outlet communicating the inside of said receptacle with the ambient environment. 
     
     
         11 . The method of  claim 10 , wherein a filter is interposed between said vent outlet and the ambient environment, and wherein said vent outlet further comprises a valve for opening and closing said outlet. 
     
     
         12 . An apparatus for connecting two or more unit operations of a manufacturing process for continuous processing comprising:
 (a) a valve manifold providing at least three fluid transmission channels comprising a central channel traversing said valve manifold and having an inlet end and an outlet end, a priming channel connecting a separate inlet with said central channel, and a bypass channel connecting a separate outlet with said central channel, each of said channels having at least one independently actuatable valve for independently opening or closing the channel, said central channel inlet being suitable for receiving throughput from an upstream unit operation, and said priming channel inlet being suitable for receiving fluid for filling said priming channel and said central channel,   (b) at least one surge receptacle having at least one inlet and at least one outlet, said surge receptacle inlet being connected to the central channel outlet of said valve manifold, and said at least one surge receptacle having the capacity to hold the throughput of a unit operation connected to said valve manifold central channel inlet.   
     
     
         13 . The apparatus of  claim 12 , wherein said surge receptacle further comprises a vent outlet communicating the inside of said receptacle with the ambient environment. 
     
     
         14 . The apparatus of  claim 13 , wherein a filter is interposed between said vent outlet and the ambient environment, and wherein said vent outlet further comprises a valve for opening and closing said outlet. 
     
     
         15 . The apparatus of  claim 14 , wherein the surge receptacle outlet is connected to a pump. 
     
     
         16 . The apparatus of any of  claims 12 - 15 , wherein said bypass channel outlet is connected to a bypass collection receptacle having the capacity to hold the throughput of a unit operation connected to said valve manifold central channel inlet. 
     
     
         17 . An apparatus for connecting two or more unit operations of a manufacturing process for continuous processing comprising:
 (a) a valve manifold providing at least five fluid transmission channels comprising a central channel traversing said valve manifold and having an inlet end and an outlet end, a priming channel connecting a separate inlet with said central channel, a surge receptacle inlet channel connecting a separate inlet with said central channel, a surge receptacle outlet channel connecting a separate outlet with said central channel, and a bypass channel connecting a separate outlet with said central channel, each of said five channels having at least one independently actuatable valve for independently opening or closing the channel, said central channel inlet being suitable for receiving throughput from an upstream unit operation, said priming channel inlet being suitable for receiving fluid for filling said priming channel and said central channel,   (b) at least one surge receptacle having at least one inlet and at least one outlet, wherein a surge receptacle inlet is connected to the surge receptacle outlet channel of said valve manifold, and a surge receptacle outlet is connected to the surge receptacle inlet channel of said valve manifold, and wherein said at least one surge receptacle has the capacity to hold the throughput of a unit operation connected to said valve manifold central channel inlet.   
     
     
         18 . The apparatus of  claim 17 , wherein said at least one surge receptacle further comprises a vent outlet communicating the inside of said receptacle with the ambient environment. 
     
     
         19 . The apparatus of  claim 18 , wherein a filter is interposed between said vent outlet and the ambient environment, and wherein said vent outlet further comprises a valve for opening and closing said outlet. 
     
     
         20 . The apparatus of  claim 19 , wherein the connection between said surge receptacle outlet and the surge receptacle inlet channel of said valve manifold is equipped with a pump for regulating fluid flow along said connection. 
     
     
         21 . The apparatus of any of  claims 17 - 20 , wherein said bypass channel outlet is connected to a bypass collection receptacle having the capacity to hold the throughput of a unit operation connected to said valve manifold central channel inlet. 
     
     
         22 . A continuous processing system comprising:
 (a) a first apparatus for carrying out a first unit operation comprising:
 (1) a plurality of valve modules, each module having a central channel traversing the module and a plurality of branch channels connecting the central channel with the outside of the valve module, wherein each of said channels has at least one independently actuatable valve for independently opening or closing the channel, wherein the valve of said central channel separates the inlet end of said central channel from the outlet end of said central channel, wherein at least two branch channels connect separate inlets to the valve module with the central channel on the inlet side of the central channel valve and at least two branch channels lead from the central channel on the outlet side of the central channel valve to separate outlets from the valve module; 
 (2) a plurality of solution conduits, each conduit connecting to a separate branch channel of each valve module, and 
 (3) a plurality of chromatography columns, each column having an inlet and an outlet, wherein said column inlet is connected to the inlet end of a valve module central channel and said column outlet is connected to the outlet end of the central channel of a different valve module, such that said plurality of columns is connected in series through invening valve modules and wherein the outlet of the last column in the series is connected, via an intervening valve module, to the inlet of the first column in the series; 
   (b) a second apparatus for carrying out a second unit operation comprising:
 (1) one or more incubation receptacles each having at least one inlet and at least one outlet, wherein collectively said one or more incubation receptacles has at least the capacity to hold the output of said first unit operation; 
 (2) a valve module comprising an inlet and an outlet, an inlet channel connected to said valve module inlet, and one or more inlet branch channels connecting with said inlet channel, wherein each inlet branch channel leads from said inlet channel and connects to an inlet of one of said one or more incubation receptacles, wherein each of said inlet branch channels has at least one independently actuatable valve for independently opening or closing the channel, the valve module further comprising an outlet channel leading to the valve module outlet, and one or more outlet branch channels connecting with said outlet channel, wherein each outlet branch channel leads from said outlet channel and connects to an outlet of one of said one or more incubation receptacles, wherein each of said outlet branch channels has at least one independently actuatable valve for independently opening or closing the channel; and 
   (c) a third apparatus, for connecting the output of said first apparatus to the inlet of said second apparatus, said third apparatus comprising:
 (1) a valve module providing at least five fluid transmission channels comprising a central channel traversing said valve module and having an inlet end and an outlet end, a priming channel connecting a separate inlet with said central channel, a surge receptacle inlet channel connecting a separate inlet with said central channel, a surge receptacle outlet channel connecting a separate outlet with said central channel, and a bypass channel connecting a separate outlet with said central channel, each of said five channels having at least one independently actuatable valve for independently opening or closing the channel, wherein said central channel inlet is connected to an outlet branch channel of each of said plurality of valve modules of the apparatus of said first unit operation, said priming channel inlet being suitable for receiving fluid for filling said priming channel and said central channel, 
 (2) at least one surge receptacle having at least one inlet and at least one outlet, wherein a surge receptacle inlet is connected to the surge receptacle outlet channel of said valve module, and a surge receptacle outlet is connected to the surge receptacle inlet channel of said valve module, and wherein said at least one surge receptacle has the capacity to hold the output of the apparatus of said first unit operation, and wherein said central channel outlet of said valve module is connected to the inlet of the valve module of said apparatus for carrying out said second unit operation. 
   
     
     
         23 . The system of  claim 22 , wherein said at least one surge receptacle and/or said one or more incubation receptacles further comprises a vent outlet communicating the inside of said receptacle with the ambient environment. 
     
     
         24 . The system of  claim 23 , wherein a filter is interposed between said vent outlet and the ambient environment, and wherein said vent outlet further comprises a valve for opening and closing said outlet. 
     
     
         25 . The system of  claim 24 , wherein the connection between said surge receptacle outlet and the surge receptacle inlet channel of said valve manifold is equipped with a pump for regulating fluid flow along said connection. 
     
     
         26 . The system of any of  claims 22 - 25 , wherein said bypass channel outlet is connected to a bypass collection receptacle having the capacity to hold the throughput of a unit operation connected to said valve manifold central channel inlet. 
     
     
         27 . The system of  claim 22 , wherein the plurality of valve modules of said apparatus for carrying out said first unit operation is are integrated into a master valve manifold. 
     
     
         28 . The system of  claim 27 , wherein said one or more incubation receptacles is equipped with means for mixing the contents of the receptacle. 
     
     
         29 . The system of  claim 28 , wherein said means is a rocker table. 
     
     
         30 . The system of  claim 22 , wherein any of said receptacles is a single-use receptacle.