JPWO2020168315A5

JPWO2020168315A5 -

Info

Publication number: JPWO2020168315A5
Application number: JP2021547125A
Authority: JP
Publication date: 2023-07-04

Claims

A process for producing a purified protein of interest comprising the steps of:
(a) a mammal under conditions that allow cells to secrete said protein into said liquid medium in one or more disposable perfusion bioreactors containing liquid medium over a productive culture period of at least 10 days; culturing the cells, periodically or continuously during said productive culture period, one or more fresh sterile liquid media mixed simultaneously from a plurality of different concentrated media component solutions and aqueous diluents; A constant in each of the perfusion bioreactors directly related to the volume of culture added to the disposable perfusion bioreactor and removed from each of the perfusion bioreactors continuously or periodically as the volume of permeate or cell bleed. Maintaining the culture volume, the removed permeate volume is automatically and fluidly supplied from one or more disposable perfusion bioreactors to a disposable surge vessel and then to a first chromatography system, thereby producing said protein. is collected in the protein isolate fraction.

the following steps,
(b) switching said protein isolate fraction to a low pH or detergent virus inactivation system and optionally a neutralization system to obtain a virus inactivation product pool comprising said protein; , the process of claim 1.

the following steps,
(c) introducing said virus-inactivated product pool into a second chromatography system to obtain a purified product pool comprising said protein;
(d) switching said purified product pool containing said protein to an optional third chromatography system and/or virus filtration system to obtain a virus-free filtrate containing said protein;
3. The process of claim 2, further comprising (e) switching the virus-free filtrate to an ultrafiltration/diafiltration system to obtain a composition comprising a purified protein of interest.

2. The process of claim 1, wherein said protein of interest is a recombinant protein.

2. The process of claim 1, wherein said protein of interest is a therapeutic protein.

said first chromatography system, said second chromatography system, said third chromatography system, said low pH or detergent virus inactivation system, said neutralization system, said virus filtration system, or said ultrafiltration/dialysis 4. The process of claim 3, wherein one or more of the filtration systems comprise one or more disposable components.

2. The process of claim 1, wherein said mammalian cells are cultured in 2, 3, 4, 5 or 6 disposable perfusion bioreactors.

2. The process of claim 1, wherein the one or more single-use bioreactors can contain a volume of liquid medium from about 50L to about 4000L.

The fresh sterile liquid medium is added to the one or more perfusion bioreactors by injecting the plurality of different concentrated medium component solutions directly into the perfusion bioreactor in fixed ratios relative to each other, while aqueous 2. The process of claim 1, wherein diluents are also added in varying ratios to said plurality of different concentrated component solutions to maintain a constant culture volume within each perfusion bioreactor.

The fresh sterile liquid medium is added to the one or more perfusion bioreactors by injecting the plurality of different concentrated medium component solutions and the aqueous diluent directly into the perfusion bioreactor in fixed ratios relative to each other. 2. The process of claim 1, wherein the culture volume in each perfusion bioreactor is maintained constant.

The fresh sterile liquid medium is added to the one or more perfusion bioreactors for culturing by injecting the plurality of different concentrated medium component solutions and the aqueous diluent into a mixing chamber in fixed ratios relative to each other. 2. The process of claim 1, wherein the volume is kept constant, wherein the fresh sterile liquid medium is mixed simultaneously before being added to each perfusion bioreactor.

An automated controller including a detector is used to measure the fluid volume in the disposable surge vessel, and a processor changes the pump speed of the first chromatography system to a preset value in the disposable surge vessel. 2. The process of claim 1, wherein the volume range is maintained.

one or more of steps (b), (c), (d), or (e) are automatically and fluidly performed in uninterrupted flow from the previous step, and one or more of the steps 4. The process of claim 3, wherein the surge vessel is used between .

14. The method of claim 13, wherein in-line or in-vessel adjustment of pH and/or conductivity loads is performed during one or more of steps (b), (c), (d), or (e). process.

(i) a process automation system is in electronic communication with at least said one or more disposable perfusion bioreactors, disposable surge vessels, and said first chromatography system;
(ii) the process automation system houses a first set of control modules for controlling operation of at least one disposable perfusion bioreactor of the one or more disposable perfusion bioreactors;
(iii) the process automation system houses a second set of control modules for controlling operation of the supply tank;
(iv) the process automation system houses a third set of control modules for controlling operation of the collection tank;
10. The method of claim 1, wherein (v) the at least one disposable perfusion bioreactor is logically configured to be connected to one or more feed tanks, one or more collection tanks, or filter banks. process.

11. The at least one disposable perfusion bioreactor is disposed on a skid, the skid comprising a plurality of communication interfaces for electronically coupling the at least one disposable perfusion bioreactor to a plurality of portable instruments. 15. The process described in 15.

determining, by the process automation system, that the disposable surge vessel is coupled to one of the plurality of communication interfaces based on data received via the communication interface; said data indicating an identifier of said disposable surge container and a function of said disposable surge container;
Based at least in part on the identifier and the function of the disposable surge vessel, the disposable surge vessel is a collection tank and the third set of control modules is for controlling operation of the disposable surge vessel. 17. The process of claim 16, further comprising determining that there is.

determining, by the process automation system, that a mixing vessel is coupled to an additional one of the plurality of communication interfaces based on additional data received via the additional communication interface; , wherein the additional data indicates an additional identifier of the mixing vessel and an additional function of the mixing vessel;
Based at least in part on the additional identifier and the additional functionality of the mixing vessel, the mixing vessel is a supply tank and the second set of control modules is for controlling operation of the mixing vessel. 18. The process of claim 17, further comprising determining that .

2. The process of claim 1, wherein the productive culture period is at least 20 days.

An automated facility for producing a purified protein of interest, said facility comprising:
(a) one or more disposable perfusion biomass capable of containing said liquid medium under conditions that allow cultured cells to secrete said protein into said liquid medium for a productive culture period of at least 10 days; fresh sterile liquid medium in direct relation to the volume of conditioned medium removed from each of said perfusion bioreactors continuously or periodically as a volume of permeate or cell bleed during said production culture period; a disposable perfusion bioreactor adapted to fluidly receive through an inlet to each of said perfusion bioreactors;
(b) a plurality of reservoirs, each adapted to contain a concentrated medium component solution or an aqueous diluent, directly or from said plurality of reservoirs in a predetermined ratio of concentrated medium component solution and aqueous diluent; fluidly connected to the inlet to each perfusion bioreactor indirectly via any mixing vessel adapted to receive and mix them simultaneously, and any mixing vessel to the inlet to each perfusion bioreactor a plurality of reservoirs in direct fluid connection;
(c) a first disposable surge vessel (SUSV1) into which a volume of the removed permeate is automatically fluidly supplied from the one or more disposable perfusion bioreactors;
(d) a first chromatography system adapted to automatically and fluidly receive cell-free permeate from said SUSV1, whereby said protein is captured in a protein isolate fraction;
An automated facility, wherein said automated facility is controlled by a process automation system (PAS).

(e) adapted to automatically and fluidly receive said protein isolate fraction from said first chromatography system, thereby yielding a virus-inactivated product pool comprising said protein, or a detergent virus inactivation system, and optionally a neutralization system;
21. The automated facility of claim 20, further comprising: (f) a holding vessel or second disposable surge vessel adapted to receive said virus inactivation product pool.

(g) a second, adapted to fluidly receive said virus-inactivated product pool from said holding container or said second disposable surge container, thereby yielding a purified product pool comprising said protein; a chromatography system;
(h) an optional third chromatography adapted to fluidly receive said purified product pool comprising said protein from said second chromatography system, thereby yielding a virus-free filtrate comprising said protein; a system and/or a virus filtration system;
(i) adapted to fluidly receive said virus-free filtrate from said second chromatography system or from said third chromatography system and/or said virus filtration system, whereby said purified protein of interest is 22. The automated facility of claim 21 , further comprising a resulting ultrafiltration/diafiltration system.

21. The automated facility of claim 20, wherein the one or more disposable perfusion bioreactors can contain a liquid medium volume of about 50L to about 4000L.

further comprising an automated controller comprising a detector for measuring the fluid volume of SUSV1 and a processor for varying the pump speed of said first chromatography system to maintain a preset volume range of SUSV1; 21. Automated equipment according to claim 20.

said first chromatography system, said second chromatography system, said third chromatography system, said low pH or detergent virus inactivation system, said neutralization system, said virus filtration system, or said ultrafiltration/dialysis 23. The automated facility of Claim 22 , wherein one or more of the filtration systems include disposable components.

directly fluidly connected downstream of said first chromatography system;
(i) a second disposable surge container; or (ii) at least two automatically switchable alternate disposable collection containers (SUCV1 and SUCV2) adapted to receive said protein isolate fractions. further includes
(i) and (ii) receive said protein isolate fraction from said first chromatography system and fluidly feed said protein isolate fraction to said low pH or detergent virus inactivation system; 21. Automated equipment according to claim 20, adapted for

said low pH or detergent virus inactivation system, and optionally said neutralization system comprising:
(i) a second disposable surge container adapted to receive said protein isolate fraction; or (ii) at least two automatically switching adapted to receive said protein isolate fraction. including possible alternative disposable collection containers (SUCV1 and SUCV2);
22. The automated facility of claim 21, wherein virus inactivation and, if necessary, neutralization is performed in a second disposable surge vessel or in SUCV1 and SUCV2.

21. The automated equipment of claim 20, further comprising hollow fiber membranes, a series of depth filters, or a filtration cart before the permeate is automatically and fluidly supplied to the SUSV1.

22. The automated facility of claim 21, comprising in (f) a disposable surge container adapted to receive said virus inactivation product pool.

21. The automated equipment of claim 20, further comprising a heat exchanger upstream of said SUSV1.

21. The automated equipment of claim 20, further comprising a filtration system upstream of said SUSV1.

(i) said second chromatography system;
(ii) said optional third chromatography system;
(iii) the virus filtration system; and (iv) the ultrafiltration/diafiltration system;
23. The automated equipment of claim 22, automatically and fluidly connected to a preceding system, wherein a surge vessel is optionally used to regulate uninterrupted flow of material between said connected systems.

said at least one or more disposable perfusion bioreactor, SUSV1, said first chromatography system, said low pH or detergent virus inactivation system, said holding vessel or disposable surge vessel, said second chromatography system, said optional the third chromatography system and/or virus filtration system and ultrafiltration/diafiltration system of comprises a first instrument positioned in a first configuration of said purified protein of interest production line;
23. The automated facility of claim 22 , wherein a first plurality of control modules are implemented to control operation of said first equipment.

A second instrument is positioned in a second configuration of the production line for additional purified protein of interest, the second configuration of the production line comprising at least one or more disposable perfusion bioreactors, the first a chromatography system, a low pH or detergent virus inactivation system, a second chromatography system, an ultrafiltration/diafiltration system, and a plurality of mixing vessels;
wherein the second configuration of the generation line is different than the first configuration of the generation line,
a second plurality of control modules implemented to control operation of the second device;
at least one mixing vessel of the plurality of mixing vessels is included in both the first configuration and the second configuration;
said at least one mixing vessel has a first function in said first configuration and a second function in said second configuration, said second function being different than said first function 34. Automated equipment according to claim 33.

A portable filter bank comprising a plurality of filter assemblies, wherein a first filter assembly of said plurality of filter assemblies comprises a first filter and a second filter assembly of said plurality of filter assemblies comprises a second filter a portable filter bank comprising:
A production equipment control system,
monitoring pressure within the first filter assembly as material flows through the first filter assembly;
determining that the pressure in the first filter assembly is at least a threshold;
a production facility control system for sending signals to operate diverter valves coupled to the first filter assembly and the second filter assembly to allow material to flow into the second filter assembly. 23. Automated equipment according to any of claims 20-22.

The one or more disposable perfusion bioreactors can contain a liquid medium under conditions that allow cultured cells to secrete the protein into the medium for a productive culture period of at least 20 days. 21. Automated equipment according to claim 20.

21. The automated equipment of claim 20, wherein said protein of interest is a recombinant protein.

21. The automated facility of claim 20, wherein said protein of interest is a therapeutic protein.

A process for producing a purified protein drug substance comprising a protein of interest, comprising the steps of:
(a) a mammal under conditions that allow cells to secrete said protein into said liquid medium in one or more disposable perfusion bioreactors containing liquid medium over a productive culture period of at least 10 days; Culturing the cells, periodically or continuously adding fresh sterile liquid medium to one or more disposable perfusion bioreactors during said production culture period to increase the volume of permeate or cell bleed Maintaining a constant culture volume in each of the perfusion bioreactors, the volume of permeate removed is directly related to the volume of culture removed from each of the perfusion bioreactors continuously or periodically as automatically and fluidly feeding from one or more disposable perfusion bioreactors to a disposable surge vessel and then to a first chromatography system, whereby said protein is collected in a protein isolate fraction;
(b) switching said protein isolate fraction to a low pH or detergent virus inactivation system and optionally a neutralization system to obtain a virus inactivation product pool comprising said protein;
(c) introducing said virus-inactivated product pool into a second chromatography system to obtain a purified product pool comprising said protein;
(d) switching said purified product pool containing said protein to an optional third chromatography system and/or virus filtration system to obtain a virus-free filtrate containing said protein;
(e) switching said virus-free filtrate to an ultrafiltration/diafiltration system to obtain said purified protein drug substance comprising said protein of interest.

a plurality of different concentrated medium component solutions and aqueous diluents before the fresh sterile liquid medium is added to the one or more perfusion bioreactors to maintain a constant culture volume in each of the perfusion bioreactors; 40. The process of claim 39, wherein the process is simultaneously mixed from

40. The process of claim 39, wherein said protein of interest is a recombinant protein.

40. The process of claim 39, wherein said protein of interest is a therapeutic protein.

An automated facility for manufacturing a purified protein drug substance, said facility comprising:
(a) one or more disposable perfusion bioreactors capable of containing a liquid medium under conditions that allow the cultured mammalian cells to secrete the protein of interest into the medium over a productive culture period of at least 10 days; wherein fresh sterile liquid medium is directly related to the volume of conditioned medium removed from each of said perfusion bioreactors continuously or periodically as volume of permeate or cell bleed during said production culture period said disposable perfusion bioreactor adapted to be fluidly received in each of said perfusion bioreactors;
(b) a first disposable surge vessel (SUSV1) into which a volume of the removed permeate is automatically fluidly supplied from the one or more disposable perfusion bioreactors;
(c) a first chromatography system adapted to automatically and fluidly receive permeate from said SUSV1, whereby said protein is captured in a protein isolate fraction;
(d) a low pH or low pH adapted to automatically and fluidly receive said protein isolate fraction from said first chromatography system, thereby yielding a virus-inactivated product pool comprising said protein; a detergent virus inactivation system, and optionally a neutralization system;
(e) a holding vessel or disposable surge vessel adapted to receive said virus inactivation product pool;
(f) a second chromatography system adapted to fluidly receive said virus-inactivated product pool from said holding vessel or disposable surge vessel, thereby yielding a purified product pool comprising said protein;
(g) an optional third chromatography adapted to fluidly receive said purified product pool comprising said protein from said second chromatography system, thereby yielding a virus-free filtrate comprising said protein; a system and/or a virus filtration system;
(h) adapted to fluidly receive said virus-free filtrate from said second chromatography system or from said third chromatography system and/or said virus filtration system, thereby obtaining said purified protein drug substance; an ultrafiltration/diafiltration system,
An automated facility, wherein said automated facility is controlled by a process automation system (PAS).

A plurality of reservoirs, each adapted to contain a concentrated medium component solution or an aqueous diluent, receives the concentrated medium component solution and the aqueous diluent directly or from the plurality of reservoirs in a predetermined ratio and mixes them simultaneously. 44. The automated facility of claim 43, which is indirectly fluidly connected to the perfusion bioreactor via any mixing vessel adapted to do so, and any mixing vessel is directly fluidly connected to the perfusion bioreactor.

44. The automated facility of claim 43, wherein said protein of interest is a recombinant protein.

44. The automated facility of claim 43, wherein said protein of interest is a therapeutic protein.

44. The automated facility of any of claims 20-22 or 43, wherein the facility is configured to operate in a continuous fashion.

40. The process of any of claims 1-3, claim 6, or claim 39, wherein said process is carried out in a continuous fashion.

40. The process of any of claims 1-3, 6, or 39, wherein the first chromatography system is disinfected prior to use with a chemical disinfectant solution containing peracetic acid.

Claim 3 or claim 39, wherein the ultrafiltration/diafiltration system comprises a single pass tangential flow filtration (SPTFF), the working pressure of the SPTFF being controlled in the range of about 0.25 psi to about 60 psi. Described process.

40. The ultrafiltration/diafiltration system of claim 3 or claim 39, wherein the ultrafiltration/diafiltration system comprises an in-line depth filtration (ILDF), the operating pressure of the ILDF being controlled in the range of about 0.25 psi to about 60 psi. process.