Single use bioreactor
A single-use bioreactor is provided. The single-use bioreactor may include a bioprocess container, a shell, at least one agitator, at least one sparger, at least one gas filter inlet port for the sparger(s) and headspace overlay, at least one fill port, at least one harvest port, at least one sample port, and at least one probe. In examples, at least one controller may monitor and control one or more parameters associated with the single-use bioreactor A method to cultivate and propagate mammalian cells is also provided. The method may include cultivating under suitable conditions and in a suitable culture medium in a first single-use bioreactor, transferring the medium containing the cells obtained by propagation from the at least one mammalian cell is into a second single-use bioreactor, transferring the medium containing the cells obtained by propagation from the at least one mammalian cell is into a third single-use bioreactor, and cultivating the cells in the third bioreactor.
1 . A bioreactor comprising:
a bioprocess container made from a liquid impermeable and flexible shape-conforming material, the bioprocess container having a bottom and at least one side wall, the bioprocess container defining a hollow enclosure for receiving culture media, the hollow enclosure containing a culture media, the bioprocess container including an interior surface configured to contact the culture media contained in the hollow enclosure and an opposite exterior surface, wherein the flexible shape-conforming material comprises a polymer film, the hollow enclosure having a volume of from about 2 L to about 50,000 L, the polymer film comprises a first hydrophilic surface;
a rigid shell defining an interior volume and having a shape configured to receive the bioprocess container therein, the rigid shell having a bottom portion defining an interior surface, the bioprocess container conforming to the interior surface of the bottom portion of the rigid shell;
a mixing device comprising a rotatable shaft that extends into the hollow enclosure of the bioprocess container and an impeller coupled to a bottom end of the rotatable shaft, the impeller comprises a second hydrophilic surface;
a drain line in fluid communication with the bioprocess container, the drain line being located at the bottom of the bioprocess container;
at least one baffle being configured to extend adjacent to the side wall of the bioprocess container in a longitudinal direction, the baffle having a shape that extends radially inward from the side wall an amount sufficient to affect fluid flow in the hollow enclosure during mixing of the culture media by the mixing device;
wherein the bioreactor further comprises at least two internal feed tubes, wherein a first feed tube is routed within the hollow enclosure to a zone around the impeller and includes a subsurface fluid outlet positioned adjacent to the impeller, the subsurface fluid outlet being associated with a fluid control device that only permits fluid to flow out of the subsurface fluid outlet and prevents fluid flow in an opposite direction, and wherein a second feed tube is positioned at a top of the bioprocess container, the second feed tube including a supersurface fluid discharge positioned above the culture media residing in the bioprocess container, the supersurface fluid discharge being located such that a fluid flowing through the fluid discharge makes direct contact with the culture media contained within the bioprocess container; and
further wherein the bioreactor is absent environmental micro-zones while in use.
2 . The bioreactor of claim 1 , wherein the baffle defines an inflatable fluid bladder, the baffle being capable of being inflated and deflated.
3 . The bioreactor of claim 1 , wherein the baffle is integral with the bioprocess container.
4 . The bioreactor of claim 1 , wherein the baffle is configured to be placed outside the hollow enclosure and wherein the side wall of the bioprocess container conforms around the shape of the baffle.
5 . The bioreactor of claim 1 , wherein the bioreactor includes from about two to about six baffles, the baffles being spaced around a circumference of the hollow enclosure of the bioprocess container.
6 . The bioreactor of claim 1 , wherein the bioprocess container has a diameter and wherein the baffle extends radially inward a distance of from about 3% to about 20% of the diameter of the bioprocess container.
7 . The bioreactor of claim 1 , wherein the baffle is made from a flexible polymer film.
8 . The bioreactor of claim 1 , wherein a diameter ratio of the impeller to the bioprocess container is from about 0.35 to about 0.55.
9 . The bioreactor of claim 1 , wherein the bioreactor further includes a top impeller above the impeller and wherein a flow number (N q ) of the top impeller and the impeller is from about 0.4 to about 0.9.
10 . The bioreactor of claim 1 , wherein the bioreactor further comprises at least one sparger and wherein the sparger comprises a ballast sparger, the ballast sparger comprising a gas tube that has a longitudinal portion and a lateral portion, the longitudinal portion extending vertically into the hollow enclosure of the bioprocess container, the lateral portion being located at an end of the longitudinal portion below the impeller, the lateral portion defining a plurality of holes for releasing a gas into the culture media contained within the bioprocess container.
11 . The bioreactor of claim 1 , further comprising a controller in communication with at least one sensor, the controller being configured to receive information from the at least one sensor and, based on the information, to control a fluid supply for varying a flow rate of a fluid from the fluid supply into the hollow enclosure of the bioprocess container for maintaining at least one parameter of the culture media contained within the hollow enclosure within preset limits.
12 . The bioreactor of claim 11 , wherein the fluid supply comprises a carbon dioxide gas supply in fluid communication with the bioprocess container and a liquid alkali supply also in fluid communication with the bioprocess container, the at least one sensor comprising a pH sensor and wherein the controller is configured to regulate pH levels of the culture media within preset limits by adding amounts of carbon dioxide gas from the carbon dioxide gas supply for selectively lowering the pH or by adding amounts of an alkali from the liquid alkali supply for selectively increasing the pH.
13 . The bioreactor of claim 11 , wherein the fluid supply comprises an oxygen gas supply and wherein the at least one sensor comprises a dissolved oxygen sensor and wherein the controller regulates dissolved oxygen levels within the culture media within present limits by periodically adding amounts of oxygen gas from the oxygen gas supply to the culture media within the hollow enclosure of the bioprocess container based on information received from the dissolved oxygen sensor.
14 . The bioreactor of claim 11 , wherein the fluid supply comprises a carbon dioxide gas supply and wherein the at least one sensor comprises a dissolved carbon dioxide sensor and wherein the controller regulates dissolved carbon dioxide levels within the culture media within present limits by periodically adding amounts of carbon dioxide gas from the carbon dioxide gas supply to the culture media within the hollow enclosure of the bioprocess container based on information received from the dissolved carbon dioxide sensor.
15 . The bioreactor of claim 12 , further comprising a thermal jacket surrounding the bioprocess container, the thermal jacket being in fluid communication with at least one of a heated fluid or a chilled fluid, the bioreactor further comprising a temperature sensor for sensing a temperature of the culture media contained within the bioprocess container, the temperature sensor being in communication with the controller, and wherein the controller is configured to receive information from the temperature sensor and, based on the information, control flow of a fluid into the thermal jacket for increasing or decreasing the temperature of the culture media contained in the bioprocess container for maintaining the culture media within preset temperature limits.
16 . The bioreactor of claim 11 , wherein the at least one sensor comprises a pH sensor and a dissolved oxygen sensor that are both in communication with the controller and wherein the controller receives information from the pH sensor and the dissolved oxygen sensor and controls a flow of different fluids into the bioprocess container for maintaining pH levels and dissolved oxygen levels of the culture media contained within the bioprocess container within preset limits.
17 . The bioreactor of claim 1 , wherein the bioreactor includes a top impeller above the impeller and wherein the top impeller forms a circumference when rotated and wherein the supersurface fluid discharge of the second feed tube is positioned above the circumference of the top impeller such that fluids flowing through the fluid discharge contact the culture media within the circumference.
18 . The bioreactor of claim 1 , wherein the interior surface of the polymer film comprises a modified polyolefin.