Bioreactor for rna in vitro transcription

Bioreactor (1) according to one of the preceding claims, wherein the movement of the magnetic particles and/or the DNA magnetic particles is configured to avoid sedimentation of the particles and/or to keep the particles free-floating.

Bioreactor (1) according to one of the preceding claims, wherein the magnet unit (3) is configured to rotate around the longitudinal axis of the reaction vessel (2), and wherein a rotation direction of the magnet unit (3) is switchable during mixing.

Bioreactor (1) according to one of the preceding claims, wherein the magnet unit (3) comprises a magnetic ring (31), and wherein the magnetic ring (31) is designed to surround the reaction vessel (2).

Bioreactor (1) according to the preceding claim, wherein the magnetic ring (31) comprises at least a first rod (320) and a second rod (322) extending from an inner circumference (34) of the magnetic ring (31) to a centre (33) of the magnetic ring (31), so that free ends (321, 323) of the first and second rod (320, 322) face each other.

Bioreactor (1) according to the preceding claim, wherein the free end (321) of first rod (320) comprises a magnet with an N pole and the free end (323) of the second rod (322) comprises a magnet with an S pole.

Bioreactor (1) according to the preceding claim, wherein the magnetic ring (31) is arranged in a housing (352) having cooling means.

Bioreactor (1) according to one of the preceding claims, wherein the magnet unit (3) further comprises a first driving means (36) configured to rotate the magnetic ring (31) and a second driving means (37) configured to move the magnetic ring (31) in the vertical direction.

Bioreactor (1) according to one of the preceding claims, wherein the reaction vessel (2) is paramagnetic or is configured to allow penetration of a magnetic field for withholding magnetic particles and DNA magnetic particles on the reaction vessel wall.

Bioreactor (1) according to one of the preceding claims, wherein the magnet unit (3) is configured to be periodically activated to mix the magnetic particles or the DNA magnetic particles.

Bioreactor (1) according to one of the preceding claims, wherein the magnet unit (3) is configured to be activated to capture the DNA magnetic particles between two subsequent RNA in vitro transcriptions on the same DNA templates.

Bioreactor (1) according to one of the preceding claims, wherein the magnet unit (3) is configured to be activated to remove the DNA magnetic particles to clean the reaction vessel.

Bioreactor (1) according to one of the preceding claims, wherein there are no mechanical motion introducing means for the DNA magnetic particles and/or the reaction vessel (2).

Bioreactor (1) according to one of the preceding claims, wherein the reaction vessel (2) comprises at least one flow breaker (4) arranged at least partially along an inner surface (21) of the reaction vessel (2) in a longitudinal direction of the reaction vessel (2).

Bioreactor (1) according to the preceding claim, wherein the reaction vessel (2) comprises two flow breakers (4) spaced apart from each other in a radial direction of the reaction vessel (2).

Bioreactor (1) according to the preceding claim, wherein the rib-shaped flow breaker (4) comprises a T- or L shaped cross section.

Bioreactor (1) according to one of the preceding claims, wherein a temperature element (5) is positioned between the inner surface (21) and the outer surface (23) of the reaction vessel (2) for adjusting a temperature of the reaction vessel (2).

Bioreactor (1) according to the preceding claim, wherein the temperature element (5) comprises a heat exchange channel (51) at least partially helically surrounding the reaction vessel (2) in a radial direction of the reaction vessel (2).

Bioreactor (1) according to the preceding claim, wherein the heat exchange channel (51) comprises a first end (52) and a second end (53), wherein the first end (52) is arranged at a top portion of the reaction vessel (2) and the second end (53) is positioned at a bottom portion of the reaction vessel (2).

Bioreactor (1) according to the preceding claim, wherein the heating wire (54) is at least partially integrated in an outer surface of the reaction vessel (2) or at least partially coated on the outer surface of the reaction vessel (2).

Bioreactor (1) according to one of the preceding claims, wherein the reaction vessel (2) is configured for an uptake of at least 20 ml of fluid, preferably 20 ml to 100 ml or 20 ml to 50 ml of fluid.

Bioreactor (1) according to one of the preceding claims, wherein the IVT master mix comprises ribonucleoside triphosphates and DNA dependent RNA polymerase.

Bioreactor (1) according to one of the preceding claims, wherein the DNA immobilization buffer comprises DNA templates and salt containing buffers.

Bioreactor (1) according to one of the preceding claims, wherein the DNA templates are linear double stranded DNA templates and preferably PCR amplified DNA templates.

Bioreactor (1) according to one of the preceding claims, wherein the magnetic particles are magnetic beads and preferably streptavidin magnetic beads or chemically functionalized magnetic beads.

Bioreactor (1) according to one of the preceding claims, wherein an inner surface of the reaction vessel (2) has a Ra value of Ra<=0.8 and preferably Ra<=0.6.

Bioreactor (1) according to the preceding claim, wherein the reaction vessel (2) comprises a port (24) at a bottom of the reaction vessel (2) for supplying and/or removing medium into/out of the reaction vessel (2), and wherein the port (24) is connectable to a valve means (60).

Bioreactor (1) according to the preceding claim, wherein the valve means (60) comprises a magnetic trap (61), and wherein the magnetic trap (61) is configured to catch magnetic particles and DNA magnetic particles.

Bioreactor (1) according to the preceding claim, wherein the magnetic trap (61) comprises an electromagnet or magnetisable spheres or a magnetisable ring and/or semi-permeable filters.

Bioreactor (1) according to the preceding claim, wherein the port (24) is positioned at the lowermost point of the reaction vessel (2).

Bioreactor (1) according to one of the preceding claims, further comprising a multi position valve (62) positioned downstream the magnetic trap and configured to direct a cleaning gas or cleaning fluid through the port (24) to remove magnetic particles and DNA magnetic particles from the port (24).

Bioreactor (1) according to the preceding claim, wherein the multi position valve (62) is configured to direct a process gas or process fluid into the reaction vessel (2) to mix the DNA magnetic particles.

Bioreactor (1) according to one of the preceding claims, wherein the bioreactor comprises at least a first leg (25) and a second leg (26) vertically supporting the bioreactor, wherein the first leg (25) comprises a first conduit (251) and the second leg (26) comprises a second conduit (261), wherein the first conduit (251) is configured to be in fluid communication with the valve means (60) and the second conduit (261) is configured to be in fluid communication with the second end (53) of the heat exchange channel (51) of the temperature element (5).

Bioreactor (1) according to one of the preceding claims, further comprising an exit port (7) connected to at least one of an exhaust duct (73) and a waste channel (74), and, optionally, an exit flow cell (72) arranged downstream the exit port (7).

Bioreactor (1) according to one of the preceding claims, further comprising a Hall sensor (63) positioned downstream the magnetic trap (61) and configured to detect magnetic fields emerging from magnetic particles or DNA magnetic particles.

Bioreactor (1) according to one of the preceding claims, wherein the reaction vessel (2) comprises Titan.

Bioreactor (1) according to one of the preceding claims, further comprising a filter element, preferably a single use filter, at the port (24) for withholding the magnetic particles in the reaction vessel (2), wherein the pores of the filter element are, preferably, of the order of 1 μm, or more preferably, have sub-micron size between 0.1 μm and 0.9 μm.

Bioreactor (1) according to one of the preceding claims, wherein the temperature element (5) is configured to adjust the reaction vessel temperature to a transcription temperature of 20 to 37° C. and preferably also to a cleaning temperature of 75 to 85° C.

Bioreactor (1) according to one of the preceding claims, wherein the valve means (60) further comprises a flow cell (64) arranged downstream the port (24).

Bioreactor (1) according to one of the preceding claims, wherein the reaction vessel (2) is further configured to hold at least one of the following elements:

• a buffer suitable for RNA in vitro transcription, a cap analogue, modified ribonucleoside triphosphates, a ribonuclease inhibitor, a pyrophosphatase, MgCl2, an antioxidant, a polyamine and a solution for cleaning and/or sanitizing.

Bioreactor (1) according to one of the preceding claims, wherein the reaction vessel (2) is further configured to hold at least one means for measuring and/or adjusting pH, salt concentration, magnesium concentration, phosphate concentration, temperature, pressure, flow velocity, RNA concentration and/or ribonucleotide triphosphate concentration.

Bioreactor (1) according to one of the preceding claims, wherein the bioreactor operates in batch, semi batch or in a repeated batch mode or in a semi-continuous or continuous mode.