Standard Operating Procedure of Applikon Bioreactor: From Assembly to Sterilization

Standard Operating Procedure : Applikon Bioreactor SOP

Target Audience: Researchers and technicians new to bioreactor systems
Purpose: To provide a detailed operational guide for setup, cleaning, sterilization, and sensor calibration of a typical Applikon bioreactor.

 

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1. Overview and Equipment Configuration

This SOP outlines the complete handling of the Applikon bioreactor system for general laboratory use. The setup includes:

• Impeller types: Rush and Marine
• Ports: DO and pH probes, sparger, overlay line, feeding/sampling ports, harvest line, condensing line
• Gas regulators: Air, CO₂ (note: pH controlled via Air only in some cases)

 

2. Cleaning Protocol

Required materials: sponge (non-detergent), hot water, distilled water (DW)

1. Disassemble impeller and sparger carefully. Do not remove bolts entirely.
2. Remove debris from sparger (notably prone to cell adhesion).
3. Clean vessel and lid thoroughly. Inspect O-rings for damage.
4. Rinse all parts 3–4 times with DW. Avoid detergent due to endotoxin risk.
5. Air-dry on clean tissue paper before reassembly.

 

3. Autoclave Guidelines

Assemble the reactor loosely to avoid pressure damage. Use 125°C for 25 minutes. Perform double-autoclaving:

• First autoclave: partial assembly (no media)
• Second autoclave: after full line assembly before culture start

 

4. Bioreactor Assembly

Use Masterflex silicon tubing (size 16/14) for all lines:

• Sampling line: split into two, connected inside the vessel
• Feeding line: two lines joined at same port as sampling
• Harvest line: securely clamped
• Overlay and condensing lines installed with air filters

Ensure couplers and cable ties are autoclaved if preconnected.

 

5. Sensor Calibration

pH Sensor:

• 25°C input required
• Calibrate with pH 4.01 and 7.00 buffers

DO Sensor:

• Handle gently with hot water rinse
• Store with sealed cap until calibration

Both probes should be autoclaved with the assembled lid (2nd autoclave only).

 

6. Final Checks and Notes

• Connect venting bottles to regulate vessel pressure
• Use autoclaved filters on all exterior connections
• Clamp all exposed tubing ends; use >2 clamps when necessary
• Optionally implement YSI Analyzer for media nutrient analysis (e.g., Glc, Lac, Gln, Glu)

 

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This SOP is written for general educational purposes and provides practical reference for labs using bioreactors for the first time.

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