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Bioreactors
Industrial Lecture Series #3
What is a Bioreactor?
An apparatus for growing organisms (yeast, bacteria, or animal cells) under controlled conditions. Used in industrial processes to produce pharmaceuticals, vaccines, or antibodies Also used to convert raw materials into useful byproducts such as in the bioconversion of corn into ethanol.
What is a Bioreactor?
Bioreactors supply a homogeneous (same throughout) environment by constantly stirring the contents. Bioreactors give the cells a controlled environment by ensuring the same temperature, pH, and oxygen levels.
Types of Bioreactors
Batch: Media and cells are added to the reactor and it is run until a predetermined set point (i.e. time, concentration). The bioreactor has a constant volume (the initial volume). Fed-Batch: The bioreactor is a batch process in the beginning and after a certain point a feed input is introduced and the volume of the vessel increases. Continuous: The bioreactor starts with an initial volume and media is constantly introduced and product is constantly taken out. The inputs and outputs are at the same rate, so the volume always remains the same.
Types of Bioreactors
Bioreactors come in many different sizes and configurations.
Bioreactor Components
Bioreactors consist of:
Vessel Agitator Sparger Baffles Probes Temperature Dissolved Oxygen pH Pressure Gauge Cooling Jacket Ports for input and output of material Condenser Back Pressure Valve Inlet Filters and Exhaust Filters Valves Load Cell
Bioreactor - Vessel
The bioreactor vessel is a container which holds the media and the cells Vessels can be made of glass, stainless steel, or a durable plastic. The durable plastic vessels are disposable (single-use). All the additional parts of a bioreactor connect to the vessel. The top of the vessel is called a head plate. On glass reactors, most additional parts are located on the head plate.
Bioreactor - Vessel
Head Plate
Vessel
Bioreactor - Agitator
An agitator is required to mix the contents in the vessel to ensure a homogeneous environment. Agitators consist of a shaft and impellers. Mixing of the bioreactor is crucial in order to supply nutrients and oxygen to the culture and to maintain a constant pH and temperature.
Bioreactor Agitators
Impellers come in many different shapes and sizes depending on what type of mixing is needed.
Bioreactor Agitators
Agitator Shaft
Impellers
Bioreactor - Sparger
The sparger is an apparatus used to introduce gasses into the vessel. Spargers are located at the bottom of the vessel and consist of a tube with tiny holes for the gas to escape through into the culture. The gas coming out of the sparger helps to aerate and mix the contents in the vessel, as well as supply oxygen to the cells.
Bioreactor - Sparger
Ring Sparger
Micro Sparger
Bioreactor - Baffles
Baffles are obstructions on the side of the vessel that generate turbulence in the flow of the culture. Baffles are made out of stainless steel and are welded to the inside of the vessel. Baffles help to mix the culture by creating a more turbulent flow.
Bioreactor - Baffles
Baffles obstruct the flow in the vessel to help mix the contents.
Baffle
Bioreactor - Probes
Bioreactors require probes to monitor the culture in the vessel. The probes are found at different locations on the vessel: head plate, top probe belt, bottom probe belt. Useful probes include temperature, pH, DO (dissolved oxygen), and CO2
Bioreactor - Probes
Probe Belt
Probes on head plate
Probes
Bioreactor - Ports
Bioreactors require addition ports, where material is either introduced or removed from the vessel. Ports are needed to add the media (media port), cells (inoculation port), and nutrients (feed ports). Ports are also used for the addition of acid and base for pH control. A sample port is also located on each vessel to remove culture for off-line investigation.
Bioreactor - Ports
Ports
Bioreactor - Condenser
A condenser is an apparatus that captures saturated air leaving the vessel. The condenser is cooler than the saturated air allowing it to condense on the surface and return to the vessel as a liquid. Condensers help to minimize the loss of material out of the reactor due to evaporation.
Bioreactor - Condenser
Bioreactor - Filters
Bioreactors need inlet filters to ensure the gasses entering the vessel are sterile. They require exhaust filters to keep the reactor sterile and allow gas to escape to regulate pressure. Filters require a filter housing a stainless steel cabinet to hold and sterilize the filter.
Bioreactor - Filters
Bioreactor - Valves
Valves are used to control the flow of either gas or liquid Bioreactors use many different types of valves.
Manual Valves open and close the valves with your hand. Pneumatic Valves automatic valve which opens and closes with the use of high pressured air (instrument air). Steam Lock Valves (so called block and bleed valves) a two valve assembly so you can allow a fluid to flow through a portion of the valve and then close and use the other portion of the valve to sterilize the
Bioreactor - Valves
Valves are used at many different locations on the bioreactor
Sample valve where a sample from the bioreactor can be obtained. Harvest valve location where the tank can be drained for harvest. Ports contain an automatic steam lock valve, so liquid can be added to the vessel (the port can be sterilized after use to prevent contamination)
Bioreactor - Valves
Manual Valve
Bioreactor Utilities
Bioreactors require utilities: Gasses Air Oxygen Carbon Dioxide Nitrogen Clean Steam - for sterilization of the reactor (condensed clean steam equals WFI (water for injection) that is clean enough to be injected) Coolant (cold water or glycol) for cooling jacket