Integrated Continuous Isolation of Fluid Streams From Sterile Process Vessels

Abstract

Provided herein are isolation processes and the associated hardware to allow fluid streams to be isolated from a sterilized system (e.g., a sterile process vessel) that contains a sterile process. The isolation processes described herein allow for continuous removal of fluid streams (e.g., waste streams, liquid containing recombinant therapeutic proteins) from a sterilized system (e.g., a biological manufacturing system), which provides for less manual manipulation of the sterilized system and a decreased risk of contaminating the sterilized system.

What is claimed is: 
     
         1 . A method of inhibiting contamination of a sterilized system, the method comprising
 providing a system comprising first vessel, wherein the first vessel comprises a liquid;   flowing a first volume of the liquid out of the first vessel and through a volume of sterilizing gas and into a second vessel.   
     
     
         2 . The method of  claim 1 , wherein the volume of sterilizing gas is disposed within a headspace of the second vessel. 
     
     
         3 . The method of  claim 2 , wherein the second vessel comprises
 (i) a fluid inlet configured such that the volume of liquid entering the second vessel passes through the head space,   (ii) a fluid outlet configured such that liquid exiting the second vessel is flowed from below the sterilizing gas-filled headspace,   (iii) at least one gas inlet; and   (iv) at least one gas outlet,   wherein the fluid inlet is in fluid communication with the first vessel.   
     
     
         4 . The method of  claim 3 , wherein the first vessel comprises
 a fluid outlet in fluid communication with a fluid inlet of the second vessel.   
     
     
         5 . The method of  claim 1 , wherein during the process the second vessel is at least partially filled with a liquid. 
     
     
         6 . The method of  claim 2 , wherein the sterilizing gas is sparged into the second vessel or introduced directly into the head space of the second vessel. 
     
     
         7 . The method of  claim 1 , wherein the first vessel is a sterilized vessel. 
     
     
         8 . The method of  claim 1 , wherein the sterilized system is a biological manufacturing system or a pharmaceutical manufacturing system. 
     
     
         9 . The method of  claim 8 , wherein the first vessel is a bioreactor, a chromatography system, a microfiltration (MF) system, or an ultrafiltration/diafiltration (UF/DF) system. 
     
     
         10 . The method of  claim 1 , wherein the liquid in the first vessel comprises a cell comprising a therapeutic protein. 
     
     
         11 . The method of  claim 1 , wherein the liquid in the first vessel comprises a recombinant therapeutic protein. 
     
     
         12 . The method of  claim 11 , wherein the recombinant protein is secreted from a cell or is not secreted from a cell. 
     
     
         13 . The method of  claim 1 , wherein the first volume of the liquid comprises a recombinant therapeutic protein. 
     
     
         14 . The method of  claim 1 , wherein the liquid in the first vessel does not comprise a recombinant therapeutic protein. 
     
     
         15 . The method of  claim 1 , wherein the first volume does not comprise a recombinant therapeutic protein. 
     
     
         16 . The method of  claim 1 , wherein the liquid in the first vessel comprises fermentation by-products. 
     
     
         17 . The method of  claim 13 , further comprising:
 (i) flowing a second volume of liquid from the second vessel into a first multi-column chromatography system (MCCS1);   (ii) capturing said recombinant therapeutic protein in the liquid culture medium using the MCCS1, wherein the eluate of the MCCS1 containing the recombinant therapeutic protein is continuously fed into a second multi-column chromatography system (MCCS2); and   (iii) purifying and polishing the recombinant therapeutic protein using the MCCS2, wherein the eluate from the MCCS2 is a recombinant therapeutic protein; and   wherein the process is integrated and runs continuously from said first vessel to the eluate from the MCCS2 that is the recombinant therapeutic protein.   
     
     
         18 . The method of  claim 13 , further comprising flowing a second volume of liquid from the second vessel into an apparatus for purifying and polishing the recombinant protein. 
     
     
         19 . The method of  claim 17 , wherein the second volume of liquid comprises a recombinant protein. 
     
     
         20 . The method of  claim 11 , wherein the recombinant therapeutic protein is an antibody or antibody fragment, an enzyme, an engineered protein, or an immunogenic protein or protein fragment. 
     
     
         21 . The method of  claim 1 , wherein the sterilizing gas is selected from the group consisting of ozone, ethylene oxide, nitrogen dioxide, or vaporized hydrogen peroxide. 
     
     
         22 . The method of  claim 1 , wherein the first volume of liquid is a waste stream. 
     
     
         23 . A system for isolating sterile process streams from non-sterile environments, comprising:
 a first vessel comprising a fluid outlet; and   at least one second vessel comprising:   (i) a fluid inlet in fluid communication with the fluid outlet of the first vessel and configured such that fluid entering the second vessel passes through a sterilizing-gas filled head space within the second vessel,   (ii) a fluid outlet configured such that fluid exiting second vessel is removed from below the sterilizing gas-filled headspace within the second vessel,   (iii) at least one gas inlet; and   (iv) at least one gas outlet.   
     
     
         24 . The system of  claim 23 , wherein the first vessel is a component of a biological manufacturing system. 
     
     
         25 . The system of  claim 24 , wherein the first vessel is a fluid conduit, a bioreactor, chromatography system, a microfiltration (MF) system, or a ultrafiltration/diafiltration (UF/DF) system. 
     
     
         26 . The system of  claim 25 , wherein the bioreactor is a production bioreactor or a seed bioreactor. 
     
     
         27 . The system of  claim 25 , wherein the bioreactor is a perfusion bioreactor or a batch-fed bioreactor. 
     
     
         28 . The system of  claim 23 , further comprising a fluid conduit disposed between the first vessel and the second vessel. 
     
     
         29 . The system of  claim 28 , further comprising a filter disposed in the fluid conduit between the first vessel and the second vessel and configured to remove particulate matter from the fluid in the fluid conduit. 
     
     
         30 . The system of  claim 28 , further comprising a pump system disposed in the fluid conduit. 
     
     
         31 . The system of  claim 30 , wherein the pump system comprises a pump configured to remove a volume of fluid from the vessel outlet and flow the volume into the fluid inlet of the second vessel. 
     
     
         32 . The system of  claim 23 , further comprising a pump system in fluid communication with the fluid outlet of the second vessel. 
     
     
         33 . The system of  claim 31 , further comprising a filter disposed in the fluid conduit between the first vessel and the pump, and configured to remove particulate matter from the fluid present in the fluid conduit. 
     
     
         34 . The system of  claim 23 , wherein the first vessel and the second vessel are disposed on a skid. 
     
     
         35 . The system of  claim 23 , wherein the sterilizing gas is selected from the group consisting of ozone, ethylene oxide, nitrogen dioxide, or vaporized hydrogen peroxide. 
     
     
         36 . The system of  claim 23 , wherein at least one gas outlet is configured to continuously or periodically vent gas from the second vessel. 
     
     
         37 . The system of  claim 23 , wherein at least one gas outlet is in gas communication with an ozone destruction unit. 
     
     
         38 . The system of  claim 23 , wherein at least one gas inlet is in gas communication with a system for generating or delivering a sterilizing gas, or for generating and delivering a sterilizing gas. 
     
     
         39 . The system of  claim 38 , wherein the system for generating or delivering a sterilizing gas is an ozone generation or delivery system, or an ozone generation and delivery system. 
     
     
         40 . The system of  claim 23 , wherein at least one gas inlet is connected to one or more gas sparging elements which permit gas to be emitted into the second vessel. 
     
     
         41 . The system of  claim 40 , wherein the gas sparging element is a filter, an open pipe or a frit. 
     
     
         42 . The system of  claim 23 , further comprising a dissolved gas probe. 
     
     
         43 . The system of  claim 23 , further comprising a sensor for monitoring the sterilizing gas concentration within the headspace of the second vessel. 
     
     
         44 . The system of  claim 23 , wherein the second vessel fluid outlet is in fluid communication with an apparatus for purifying and polishing a recombinant protein. 
     
     
         45 . The system of  claim 23 , wherein the vessel is a sterilized vessel.