US-12624332-B2 - Automated integrated continuous system and bioprocess for producing therapeutic protein

US12624332B2US 12624332 B2US12624332 B2US 12624332B2US-12624332-B2

Abstract

The present invention relates to an automated integrated continuous bioprocess system and bioprocess that are capable of continuously producing therapeutic protein in an uninterrupted manner and scalable from laboratory to manufacturing scale. The present invention provides an automated integrated continuous bioprocess system and bioprocess for producing therapeutic protein, in which the system and process is controlled with one or more control system selected from supervisory control and data acquisition (SCADA) control system ( 110 ), proportional integral derivative (PID), programmable logic circuit (PLC), industrial PC (IPC), distributed control system (DCS), message relaying system and automated UPLC/HPLC sampling for online testing to run the system and process and in an uninterrupted manner and continuously produce therapeutic protein.

Inventors

Hrishikesh BARTAKE
Himanshu Gadgil
Abir BANERJEE
Harshita Londhe
Abijar BHORI
Abhijit BUTTI
Samir Varma
Rohan GODSE
Veerendra REDDY

Assignees

ENZENE BIOSCIENCES LIMITED

Dates

Publication Date: 20260512
Application Date: 20190401
Priority Date: 20190125

Claims (7)

1 . An automated integrated continuous bioprocess system ( 100 ) for producing therapeutic protein comprising: a bioreactor ( 103 ) to culture mammalian cells capable of producing the therapeutic protein in culture medium, the bioreactor ( 103 ) comprises alternating tangential flow (ATF) filter ( 109 ) to collect a harvest comprising the therapeutic protein secreted into the culture medium; a first chromatography system ( 119 ) comprising one or more affinity chromatography column(s) connected to the ATF filter ( 109 ) without any intermediate hold vessel to purify the harvest comprising the therapeutic protein and provide protein A eluate; a viral inactivation system ( 126 ) including a viral inactivation vessel ( 128 ) with a pH probe connected to a transmitter for measuring the pH of the protein A elute connected to the first chromatography system ( 119 ) to collect the protein A eluate and inactivate viruses that may be present in the protein A eluate, and the viral inactivation vessel ( 128 ) configured to automatically adjust pH of the protein A eluate to obtain a virus inactivated, neutralized, and filtered protein A eluate; a collection vessel ( 136 ) connected with the viral inactivation vessel ( 128 ) through one or more filters to receive the virus inactivated, neutralized, and filtered protein A eluate, wherein said one or more filter(s) ( 224 ) and ( 226 ) are configured to remove impurities as precipitates from the virus inactivated, neutralized, and filtered protein A eluate received from the viral inactivation vessel; a second chromatography system ( 137 ) comprising one or more multimodal anion exchange column(s) and one or more cation exchange column(s) connected with the collection vessel ( 136 ) to receive the virus inactivated, neutralized, and filtered protein A eluate from the collection vessel ( 136 ) and to provide purified protein; one or more of solenoid or pneumatic pinch valves ( 114 ), sensors, air bubble sensors, pressure sensors and load cell sensors for creating a feedback loop in order to maintain fluid flow using tubes between various components of the system, avoid formation of air bubbles and regulate flow of liquid; cleaning in place (CIP) system ( 120 ) comprising a plurality of reservoirs of cleaning solutions and a plurality of valves for periodic cleaning of the first chromatography system and/or second chromatography system, viral inactivation vessel, collection vessel, and tubes; one or more control systems selected from supervisory control and data acquisition (SCADA) control system ( 110 ), proportional integral derivative (PID); programmable logic circuit (PLC), Industrial PC (IPC), distributed control system (DCS); Input-Output Modules or IO Boxes ( 130 ) and ( 132 ) operably connected with the first chromatography system ( 119 ); the viral inactivating system ( 126 ); the collection vessel ( 136 ); and the second chromatography system ( 137 ).
2 . The automated integrated continuous bioprocess system as claimed in claim 1 , further comprising a surge bag ( 116 ) connected with the bioreactor through a valve ( 114 ).
3 . The automated integrated continuous bioprocess system as claimed in claim 1 , wherein the viral inactivation system ( 126 ) comprises a viral inactivation (VI) vessel ( 128 ), a pH probe ( 220 ) connected to a transmitter for measuring the pH of the protein A eluate, autotitrator ( 122 ), load cell sensors ( 248 - 1 , 248 - 2 , 248 - 3 , 248 - 4 , 248 - 5 , and 248 - 6 ) to monitor the level of liquids in respective load cells, a level sensor to check level of fluid in the VI vessel, pressure sensors ( 256 - 1 ) and ( 256 - 2 ) in the fluid path, inline turbidity measuring sensor ( 254 ) to measure real time turbidity, and an air bubble sensor ( 252 ) to prevent air bubble entering into the first chromatography system and/or second chromatography system.
4 . The automated integrated continuous bioprocess system as claimed in claim 1 , wherein each of the filters between viral inactivation vessel and the collecting vessel is 0.2-0.45 micron filter.
5 . The automated integrated continuous bioprocess system as claimed in claim 1 , wherein the viral inactivation vessel, and the collection vessel are made of glass or stainless steel.
6 . The automated integrated continuous bioprocess system as claimed in claim 1 , wherein the cleaning in place (CIP) system ( 120 ) for periodic cleaning of the first chromatography system and/or second chromatography system includes cleaning of components thereof including chromatographic column(s), and inlet.
7 . The automated integrated continuous bioprocess system as claimed in claim 1 , further comprises an automated harvest sampling from bioreactor for cell count and nutrient analysis, and automated sampling at different locations for online chromatography analysis in the automated integrated continuous bioprocess.

Description

FIELD OF THE INVENTION The present invention generally relates to system for continuous bioprocess for producing therapeutic protein. Specifically, the present invention relates to an automated integrated continuous bioprocess system and bioprocess that are capable of continuously producing therapeutic protein and scalable from laboratory to manufacturing scale. BACKGROUND OF THE INVENTION Background description includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art. Identification of significance of therapeutic protein has led to new revolution in biopharmaceutical industry. However, production of therapeutic protein commonly observes their charged isoforms which largely interfere in the isolation and purification processes that are required to attain high yields and quality in the product. Conventionally, therapeutic proteins are manufactured by biopharmaceutical companies using batch processing, in which unit operations are performed and completed before the process stream moves to the next step. Recently, biopharmaceutical companies are following continuous bio-process for manufacturing of the therapeutic protein. In continuous bio-processing, the processed products are moved to the next step as each unit process is completed. Continuous bio-processing has gained a large amount of interest because of various advantages like steady-state operation, small equipment size, high volumetric productivity, streamlined process flow, low cycle time and reduced capital cost. Various approaches have been attempted in the existing art for manufacturing therapeutic protein using continuous bio-process methods. However, such existing continuous bio-processes are pseudo continuous process containing standalone units, each performing their function. In such systems each unit operation has limited communication between the individual system parameters and operate mostly in isolation. Further, the current continuous processes mostly involve offline chromatography analysis with limited or no feedback to the ongoing process. As biopharmaceutical companies continue to grow, there is an unmet need to provide an automated integrated bio-process and system for manufacturing therapeutic protein that can be scalable from laboratory to manufacturing scale. OBJECTS OF THE INVENTION An object of the present invention is to provide an automated integrated continuous bioprocess system for producing therapeutic protein. Another object of the present invention is to provide an automated integrated continuous bioprocess for producing therapeutic protein using a control system with communication and programmable control function to regulate the entire process parameters using a master controller. Another object of the present invention is to provide an automated integrated continuous bioprocess for producing therapeutic protein which can be carried out in continuous and uninterrupted manner. SUMMARY OF THE INVENTION The present invention in some of the aspects relate to an automated integrated continuous bioprocess system for producing therapeutic protein in an uninterrupted manner using a control system with communication and programmable control function to regulate the entire process parameters using a master controller In an aspect the present invention provides an automated integrated continuous bioprocess system for producing a therapeutic protein comprising: a bioreactor (103) to culture mammalian cells capable of producing the therapeutic protein in the culture medium, the bioreactor (103) enabled with alternating tangential flow (ATF) filter (109) to collect a harvest comprising the protein secreted into the culture medium;a first chromatography system (119) connected to the ATF filtration system (109) of the bioreactor (103) without any intermediate hold vessel to purify the harvested recombinant therapeutic protein and provide protein A eluate;a viral inactivation system (126) including a viral inactivation vessel (128) connected to the a first chromatography system (119) to collect the protein A eluate and inactivate viruses that may be present in the eluate, and the viral inactivation vessel (128) configured to automatically adjust pH of the protein A eluate;a collection vessel (136) connected with the viral inactivation vessel (128) through one or more filters to receive the virus inactivated, neutralized, and filtered protein A eluate, wherein said one or more filter(s) (224) and (226) are configured to remove impurities in the form of precipitates from the neutralized protein A eluate received from the viral inactivation vessel;a second chromatography system (137) connected with the collection vessel (136) to receive the filtered protein A eluate from the collection vessel (136) and provide further