Bioreactor for the production of biomolecules

Abstract

The invention relates to a bioreactor (1) comprising a biomolecule production device (2) including at least one reaction tank (3) designed for synthesizing the biomolecules. The reaction tank includes a conduit (4) describing a left-handed curve extending around at least one helix axis (Ah1, Ah2) to form a succession of loops, said conduit fluidly connecting a feed inlet (5) of said at least one reaction tank to a discharge outlet (6) of said at least one reaction tank. The conduit according to the invention is rotatable about an axis of rotation (Ar1) so as to allow the flow of a liquid from one loop to another in a certain fluidic flow direction (F1) from the feed inlet to the discharge outlet when the conduit rotates about said axis of rotation. Figure to be published: 2

Description

Title of the invention: Bioreactor for the production of biomolecules Technical field

[0001] The invention falls within the field of the production of biomolecules and relates more particularly to a bioreactor comprising a device for producing biomolecules comprising at least one reaction tank designed to synthesize the biomolecules.

[0002] The invention also relates to a method for producing biomolecules in a bioreactor according to the invention. Prior art

[0003] Scientific and technological progress in recent years has made it possible to expand the use of biomolecules to biological diagnostics and therapeutic treatments.

[0004] The rise of new therapies targeting genetic machinery and protein expression opens up new opportunities for the use of biomolecules, particularly in the development of innovative therapeutic treatments, such as vaccines.

[0005] In particular, there is a strong interest in the large-scale production of messenger RNA (mRNA) vaccines, despite the numerous technical difficulties associated with its production and storage.

[0006] Document WO2022 / 261050 describes in this regard a new type of bioreactor for the continuous production of mRNA in which a reaction reservoir makes it possible to synthesize a large quantity of mRNA in vitro and in which affinity chromatography is carried out on the mRNA for its purification. The continuous synthesis of mRNA is made possible here by the use of an injection device designed to continuously inject reagents on the basis of a measurement of the concentration of the reagents in the reservoir in real time.

[0007] This type of bioreactor does not, however, allow scaling up of mRNA production since the reaction tank is sized to synthesize a certain minimum quantity of mRNA.

[0008] On the other hand, the mRNA recovered at the outlet of the reaction tank is accompanied by a remainder of reagents which did not allow the production of mRNA. Thus, if the use of affinity chromatography makes it possible to efficiently separate the mRNA from these reagents, this bioreactor consumes a very large quantity of expensive reagents, such as enzymes and dNTPs, compared to the quantity of mRNA produced. Summary of the invention

[0009] The objective of the invention is therefore to solve the aforementioned problems.

[0010] For this purpose, the subject of the invention is a bioreactor comprising a device for producing biomolecules comprising at least one reaction tank designed to synthesize the biomolecules, characterized in that said at least one reaction tank comprises a conduit describing a left curve extending around at least one helix axis to form a succession of loops, said conduit fluidly connecting a feed inlet of said at least one reaction tank to a discharge outlet of said at least one reaction tank and in that the conduit is rotatable around an axis of rotation so as to allow the flow of a liquid from one loop to another in a certain direction of fluid flow from the feed inlet to the discharge outlet when the conduit rotates around said axis of rotation.

[0011] The idea behind the invention is to sequentialize the synthesis of biomolecules in each loop of the reaction tank and therefore to produce from one to a multitude of sub-batches of biomolecules. Thus, thanks to the bioreactor according to the invention, a certain number of the sub-batches produced can be mixed to form a single batch of biomolecules at the desired scale.

[0012] The bioreactor according to the invention may also have the following particularities:

[0013] - the conduit is designed to perform successive rotations of 360° in the same direction of rotation like an Archimedes screw;

[0014] - the conduit is designed to perform successive rotations of 180° in directions of opposite rotation;

[0015] - the propeller axis has an angle of inclination relative to the horizontal allowing the storage of said liquid in the loop in which it is located between two rotations of the conduit around the axis of rotation;

[0016] - the conduit is inclined so that the supply inlet is a high point said at least one reaction tank and the discharge outlet a low point of said at least one reaction tank;

[0017] - the feed inlet comprises an upstream conduit section and in that the discharge outlet comprises a downstream conduit section, said upstream conduit section and said downstream conduit section extending longitudinally in the axis of rotation of the conduit;

[0018] - the axis of rotation is offset parallel to the propeller axis;

[0019] - the axis of rotation is angularly offset relative to the propeller axis;

[0020] - the production device comprises a plurality of connected reaction tanks fluidly in series according to said certain direction of fluid flow;

[0021] - the production device comprises a double connector designed to connect fluid specifically two reaction tanks to each other, said double connector comprising two movable parts connected respectively to the discharge outlet of one of the two reaction tanks and to the supply inlet of the other reaction tanks, said double connector comprising two fixed parts arranged between the two movable parts so as to allow rotation of the two reaction tanks relative to the fixed part and a central conduit section arranged between the two fixed parts, the production device comprising an injection device fluidically connected to the central conduit section and is adapted to inject a fluid into the central section in the fluid flow direction;

[0022] - the production device comprises a storage tank fluidly connected to the discharge outlet of the reaction tank having a volume equivalent to the volume of a plurality of loops of the tank, said storage tank being adapted to store liquid from the reaction tank.

[0023] The invention also extends to a method for producing biomolecules in a bioreactor according to the invention, characterized in that the method comprises the following steps consisting of:

[0024] a) injecting through the feed inlet a certain volume of a liquid reaction mixture, corresponding to a sub-batch, into a loop of said at least one reaction tank of the upstream production device in the fluid flow direction,

[0025] b) rotating the conduit so as to move said liquid reaction mixture into the next loop in the direction of the discharge outlet,

[0026] c) repeating steps a) and b) as many times as there are sub-batches of biomolecules to be produced in the same batch of biomolecules.

[0027] The idea of ​​the invention here is to optimize the synthesis reaction by carrying out a tailor-made supply of reagents into the reaction tank. Each loop will thus include the right amount of reagents necessary for the production of a nominal quantity of biomolecules.

[0028] Thus, the invention allows scaling up of the synthesis of biomolecules by repeating the injection of the reaction mixture and the rotation of the conduit as many times as desired.

[0029] The method according to the invention may also include the following characteristics:

[0030] - a step of stirring the conduit is carried out after step a) and before step b);

[0031] - the stirring step consists of carrying out an incomplete rotation of the conduit around the axis of rotation so as to prevent the flow of liquid from one loop to another;

[0032] - the volume of liquid reaction mixture injected in step a) is less than the volume inside a loop;

[0033] - the volume of liquid reaction mixture injected in step a) allows the passage of a gaseous fluid from the feed inlet to the discharge outlet of said at least one reaction tank;

[0034] - rotation of an upstream reaction tank in the flow direction fluidic is faster than the rotation of another downstream reaction tank in the fluidic flow direction so that a plurality of sub-batches distributed in a certain first number of successive loops (ni) of the upstream reaction tank, is distributed in a certain second number of loops (n2) of the downstream reaction tank less than said certain first number of loops, so that ni>2 and n2 <nl ;

[0035] - each rotation of the conduit is 360° in the same direction of rotation;

[0036] - each rotation of the conduit is 180° alternating the direction of rotation at each rotation ;

[0037] - the biomolecules produced are chosen from RNAs, DNAs and proteins;

[0038] - the produced biomolecules are used as a therapeutic agent. Summary presentation of the drawings

[0039] The present invention will be better understood and other advantages will appear on reading the detailed description of an embodiment taken as a non-limiting example and illustrated by the appended drawings, in which:

[0040] - [Fig.l] [Fig.l] is a schematic representation of a bioreactor according to the invention;

[0041] - [Fig.2] [Fig.2] is a schematic representation of a production device of biomolecules of the bioreactor according to the invention, according to a first exemplary embodiment of the invention;

[0042] - [Fig.3] [Fig.3] is a schematic representation of a production device of biomolecules of the bioreactor according to the invention comprising two reaction tanks in series;

[0043] - [Fig.4] [Fig.4] is a schematic representation of a production device of biomolecules of the bioreactor according to the invention, according to a second exemplary embodiment of the invention;

[0044] - [Fig.5] [Fig.5] is a diagram showing an example of implementation of the steps of the method according to the invention. Description of an embodiment

[0045] The bioreactor 1 according to the invention, as shown in [Fig.l], is designed to produce on the desired scale, a certain quantity of biomolecules such as RNA (ribonucleic acid), DNA (deoxyribonucleic acid) or proteins. These biomolecules can be used as therapeutic agents and more particularly in the manufacture of vaccines.

[0046] Scale production according to the invention consists of producing biomolecules in the form of sub-batches, a certain number of which can then be mixed to form a batch of biomolecules in the desired quantity.

[0047] The bioreactor 1 according to the invention will here comprise a production device 2 for biomolecules designed to synthesize the biomolecules using a reaction tank 3, visible in Figures 2, 3 and 4.

[0048] Structurally, the reaction tank 3 comprises a conduit 4 describing a left-hand curve extending around at least one helix axis Ahl, Ah2, to form a succession of loops or turns. By left-hand curve, we mean a curve which is not defined in a plane. As shown in [Fig.2], the conduit 4 according to the invention fluidically connects a supply inlet 5 of the reaction tank 3 to a discharge outlet 6 of the reaction tank 3. The conduit 4 is here rotatable around an axis of rotation Arl so as to allow the flow of a liquid from one loop to another according to a certain fluid flow direction Fl from the supply inlet 5 to the discharge outlet 6 when the conduit 4 rotates around the axis of rotation Arl.

[0049] Without restricting the scope of the invention, the conduit 4 may be in the form, for example, of a tube with a circular or rectangular section, a gutter, or any other shape suitable for moving a liquid from the supply inlet 5 to the discharge outlet 6.

[0050] A first injection device 7 may also be provided upstream of the supply inlet 5 in the fluid flow direction FL. The first injection device 7 will here be designed to inject a certain volume of liquid and at a certain flow rate through the supply inlet 5.

[0051] A mixing tank 8, visible in [Fig.2], may also be provided between the first injection device 7 and the feed inlet 5 to allow the intermediate storage of a liquid before its injection into the reaction tank 3.

[0052] Without restricting the scope of the invention, the propeller axis Ahl, Ah2, may have an angle of inclination a relative to the horizontal H allowing the storage of said liquid in the loop in which it is located and between two rotations of the conduit 4. This inclination therefore excludes the possibility of having a vertical propeller axis Ahl, Ah2 and therefore prevents the flow of the liquid by gravity from the supply inlet 5 to the discharge outlet 6.

[0053] The angle of inclination of the propeller axis Ahl, Ah2, may preferably be between 0° and 50° relative to the horizontal, preferably between 0° and 15°. The angle of inclination of the propeller axis Ahl, Ah2, may more preferably be 10°.

[0054] The conduit 4, and more precisely the axis of the propeller Ahl, Ah2, may also be inclined relative to the horizontal H, so that the feed inlet 5 is a high point of the reaction tank 3 and the discharge outlet 6 a low point of the reaction tank 3. This arrangement of the conduit 4 makes it possible to facilitate the flow of a fluid in the first loop of the conduit 4 and to facilitate the flow of a fluid by gravity through the discharge outlet 6. It is understood that the left curve formed by the conduit 4 prevents the gravitational flow of a liquid from one loop to another when the propeller axis Ahl, Ah2, has this inclination.

[0055] Without restricting the scope of the invention, the axis of the propeller Ahl, Ah2, may also be inclined relative to the horizontal H, so that the feed inlet 5 is a low point of the reaction tank 3 and the discharge outlet 6 a high point of the reaction tank 3 so as to raise a liquid relative to the ground.

[0056] In order to facilitate the supply of liquid via the supply inlet 5 and its evacuation via the evacuation outlet 6, the supply inlet 5 and the evacuation outlet 6 may respectively comprise an upstream conduit section 9 and a downstream conduit section 10 which extend longitudinally in the axis of rotation Arl of the conduit 4. The orientation of these sections makes it possible in particular to limit turbulence thanks to a longitudinal fluid flow and to avoid the use of a pump at the evacuation outlet 6 thanks to a gravity fluid flow.

[0057] In addition, the axis of rotation Arl of the conduit 4 may extend in the extension of the downstream 10 and upstream 9 conduit sections in order to facilitate the fluid flow.

[0058] In order to improve the synthesis yield of biomolecules, the reaction tank 3 according to the invention may be designed to agitate a liquid inside the loops. For this, the axis of rotation Arl of the conduit 4 may for example be offset parallel to the helix axis Ahl, Ah2, and / or angularly offset relative to the helix axis, as shown in [Fig.4].

[0059] In an alternative shown in [Fig.2], the rotation axis Arl may merge with the propeller axis Ahl, Ah2, in order to have a very compact reaction tank 3 and in order to limit the constraints during the rotation of the conduit 4.

[0060] The production device 2 according to the invention may comprise a storage tank 11 fluidly connected to the discharge outlet 6 of the reaction tank 3, the volume of which is equivalent to the volume of a plurality of loops of the reaction tank 3. The storage tank 11 is here designed to store a liquid coming from the reaction tank 3.

[0061] The production device 2 according to the invention may also comprise a plurality of reaction tanks 3 fluidly connected in series in the fluid flow direction Fl.

[0062] Also, in order to allow different rotation speeds for each reaction tank 3, the production device 2 will for example be equipped with a double connector 12 designed to fluidly and rotationally connect two reaction tanks 3 to each other.

[0063] The double connector 12 according to the invention comprises two movable parts 13 connected respectively to the discharge outlet 6 of one of the two reaction tanks 3 and to the supply inlet 5 of the other reaction tank 3. The double connector 12 also comprises two fixed parts 14 arranged between the two movable parts 13 so as to allow rotation of the two reaction tanks 3 relative to the fixed parts 14. The double connector 12 also comprises a central conduit section 15 arranged between the two fixed parts 14.

[0064] Thanks to the double connector 12, the production device 2 can be equipped with a second injection device 16 fluidically connected to the central section 15 of conduit so as to allow the injection of a fluid into the central section 15 in the fluid flow direction FL. The injection device 16 allows for example the addition of reagents in order to optimize the synthesis of the biomolecules.

[0065] It is obvious that the bioreactor 1 according to the invention can comprise other devices arranged one after the other in the fluid flow direction F1 following the production device 2, such as for example a purification device 17 and a device 18 for encapsulating the biomolecules.

[0066] In a first exemplary embodiment according to the invention shown in [Fig.2], the conduit 4 may be designed to perform successive rotations of 360° in the same direction of rotation in the manner of an Archimedean screw. The conduit 4 will here describe a left curve around a helix axis Ahl to form loops, here by way of example in the manner of a coil. Thus, the flow of a liquid will be from one loop to another according to the fluid flow direction Fl when the conduit 4 performs a complete rotation around the rotation axis Arl. By Archimedean screw is meant any principle allowing a liquid to be moved in translation parallel to the axis of the helix Ahl towards the discharge outlet 6.

[0067] In a second embodiment according to the invention shown in [Fig. 3], the conduit 4 may be designed to perform successive rotations of 180° in opposite directions of rotation. The conduit will in this case describe a left curve around two helix axes Ahl, Ah2, distinct to form substantially an eight, each loop of the eight being crossed by one of the two helix axes Ahl, Ah2. In this embodiment, the eight-shaped conduit 4 will preferably, but not necessarily, be oriented vertically between two rotations. By vertically oriented, we mean means that the loops of the figure eight are oriented one above the other. With this orientation, a simple rotation of 180° in one direction then in the other allows the fluid to advance towards the discharge outlet 6. This embodiment thus makes it easier to add cabling or sensors inside the conduit 4, since the conduit 4 does not perform a complete rotation sequence.

[0068] The invention also relates to a method for producing biomolecules with the bioreactor 1 described above, in particular RNAs, DNAs and proteins, for example for the manufacture of vaccines.

[0069] The succession of steps described below and represented in the diagram of [Fig.5] is an example of implementation of the method according to the invention which is in no way limiting.

[0070] A first step 100 consists of injecting through the feed inlet 5 a certain volume of a liquid reaction mixture, corresponding to a sub-batch of biomolecules to be produced, into a loop of the reaction tank 3 of the production device 2 upstream in the fluid flow direction FL. This first step may be carried out by simple gravity flow or carried out using a pump.

[0071] The volume of injected liquid reaction mixture may advantageously be less than the internal volume of a loop so as to avoid mixing of liquid between adjacent loops.

[0072] More precisely, the volume of injected liquid reaction mixture may allow the passage of a gaseous fluid from the feed inlet 5 to the discharge outlet 6 of said at least one reaction tank. This involves advantageously creating a space so that the liquid can be correctly homogenized.

[0073] A second step 200 consists of rotating the conduit 4 so as to move said liquid reaction mixture into the next loop in the direction of the discharge outlet 6. In the case of the first embodiment, a rotation of 360° will be performed, whereas in the case of the second example, a rotation of 180° will be performed.

[0074] A third step 300 consists of agitating the conduit after the first step but before the fourth step. For example, this third step will consist more particularly in carrying out an incomplete rotation of the conduit 4 around the axis of rotation Arl so as to prevent the flow of a liquid from one loop to another. For example, a rotation of less than 90° relative to the horizontal. These rotations for agitation may be alternated in both directions of rotation.

[0075] A fourth step 400 consists of repeating the first, second and third steps as many times as there are sub-batches of biomolecules to be produced in the same batch of biomolecules. In the case of the first example, the rotations to pass the liquid reaction mixture from one loop to another will always be carried out in the same direction of rotation whereas in the case of the second example, the rotations will be carried out in one direction then in the other.

[0076] In the case of using a plurality of reaction tanks 3 arranged in series, a control-command unit 19 will be able to control the rotation speed of an upstream reaction tank 3 in the fluid flow direction F1 so that its rotation speed is faster than the rotation of another reaction tank 3 downstream in the fluid flow direction F1. This speed differential makes it possible to distribute a plurality of sub-batches synthesized in a first number of successive loops (ni) of the upstream reaction tank 3, in a certain second number of loops (n2) of the downstream reaction tank 3 lower than the certain first number of loops, so that ni>2 and n2

Claims

Claims

1. Bioreactor (1) comprising a production device (2) for biomolecules comprising at least one reaction tank (3) designed to synthesize the biomolecules, characterized in that said at least one reaction tank comprises a conduit (4) describing a left curve extending around at least one helix axis (Ahl, Ah2) to form a succession of loops, said conduit fluidically connecting a feed inlet (5) of said at least one reaction tank to a discharge outlet (6) of said at least one reaction tank and in that the conduit is rotatable around an axis of rotation (Arl) so as to allow the flow of a liquid from one loop to another according to a certain fluidic flow direction (Fl) from the feed inlet to the discharge outlet when the conduit rotates around said axis of rotation.

2. Bioreactor according to claim 1, characterized in that the conduit is designed to perform successive rotations of 360° in the same direction of rotation in the manner of an Archimedes screw.

3. Bioreactor according to claim 1, characterized in that the conduit is designed to perform successive rotations of 180° in opposite directions of rotation.

4. Bioreactor according to any one of the preceding claims, characterized in that the helix axis has an angle of inclination (a) relative to the horizontal allowing the storage of said liquid in the loop in which it is located between two rotations of the conduit around the axis of rotation.

5. Bioreactor according to any one of the preceding claims, characterized in that the conduit is inclined so that the feed inlet is a high point of said at least one reaction tank and the discharge outlet a low point of said at least one reaction tank.

6. Bioreactor according to any one of the preceding claims, characterized in that the feed inlet comprises an upstream conduit section (9) and in that the discharge outlet comprises a downstream conduit section (10), said upstream conduit section and said downstream conduit section extending longitudinally in the axis of rotation of the conduit.

7. Bioreactor according to any one of claims 1 to 6, ca- characterized in that the axis of rotation is offset parallel to the propeller axis.

8. Bioreactor according to any one of claims 1 to 6, characterized in that the axis of rotation is angularly offset relative to the propeller axis.

9. Bioreactor according to any one of the preceding claims, characterized in that the production device comprises a plurality of reaction tanks fluidically connected in series along said certain fluid flow direction.

10. Bioreactor according to claim 9, characterized in that the production device comprises a double connector (12) designed to fluidically connect two reaction tanks to each other, said double connector comprising two movable parts connected respectively to the discharge outlet of one of the two reaction tanks and to the supply inlet of the other reaction tanks, said double connector comprising two fixed parts arranged between the two movable parts so as to allow rotation of the two reaction tanks relative to the fixed part and a central conduit section arranged between the two fixed parts, the production device comprising an injection device fluidically connected to the central conduit section and is designed to inject a fluid into the central section in the fluid flow direction.

11. Bioreactor according to any one of the preceding claims, characterized in that the production device comprises a storage tank (11) fluidically connected to the discharge outlet of the reaction tank whose volume is equivalent to the volume of a plurality of loops of the tank, said storage tank being adapted to store liquid from the reaction tank.

12. A method for producing biomolecules in a bioreactor according to any one of the preceding claims, characterized in that the method comprises the following steps: a) injecting (100) through the feed inlet a certain volume of a liquid reaction mixture, corresponding to a sub-batch, into a loop of said at least one reaction tank of the upstream production device in the fluid flow direction, b) rotating (200) the conduit so as to move said liquid reaction mixture into the next loop in the direction of the discharge outlet, c) repeating (400) steps a) and b) as many times as there are sub-batches of biomolecules to be produced in the same batch of biomolecules.

13. Method for producing biomolecules according to claim 12, characterized in that it comprises a step (300) of stirring the conduit carried out after step a) and before step b).

14. A method of producing biomolecules according to claim 13, characterized in that the stirring step consists of performing an incomplete rotation of the conduit around the axis of rotation so as to prevent the flow of a liquid from one loop to another.

15. A method for producing biomolecules according to any one of claims 12 to 14, characterized in that the volume of liquid reaction mixture injected in step a) is less than the internal volume of a loop.

16. Process for producing biomolecules according to any one of claims 12 to 15, characterized in that the volume of liquid reaction mixture injected in step a) allows the passage of a gaseous fluid from the feed inlet to the discharge outlet of said at least one reaction tank.

17. A method for producing biomolecules according to any one of claims 12 to 16, characterized in that the rotation of an upstream reaction tank in the fluid flow direction is faster than the rotation of another downstream reaction tank in the fluid flow direction so that a plurality of sub-batches distributed in a certain first number of successive loops (ni) of the upstream reaction tank, is distributed in a certain second number of loops (n2) of the downstream reaction tank less than said certain first number of loops, so that ni>2 and n2 <nl.

18. A method for producing biomolecules according to any one of claims 12 to 17, characterized in that each rotation of the conduit is 360° in the same direction of rotation.

19. A method of producing biomolecules according to any one of claims 12 to 17, characterized in that each rotation of the conduit is 180°, alternating the direction of rotation with each rotation.

20. Method for producing biomolecules according to any one of claims 12 to 19, characterized in that the biomolecules produced are chosen from RNAs, DNAs and proteins.

21. A method of producing biomolecules according to any one of claims 12 to 20, characterized in that the biomolecules produced are used as a therapeutic agent.

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