Standard Operating Procedure of Applikon Bioreactor: From Assembly to Sterilization

🧪 Introduction: Why a Detailed SOP for Applikon Bioreactor Matters in 2025

In modern biotech labs, where reproducibility and precision are non-negotiable, the Applikon Bioreactor stands out as a gold-standard system for small- to mid-scale bioprocessing. From microbial fermentation to mammalian cell culture, its modular design and flexible control interfaces have made it a staple in both academic and industrial settings.

As of 2025, the Applikon Bioreactor continues to evolve with advanced automation features, seamless data integration, and compatibility with GMP-compliant workflows. Its wide adoption across pharmaceutical R&D, synthetic biology, and metabolic engineering makes it essential for technicians and researchers to understand not just how to run it, and more importantly, how to do it right from day one.

This document provides a comprehensive Standard Operating Procedure (SOP) for the Applikon Bioreactor, covering everything from initial assembly and cleaning to sterilization protocols and sensor calibration. Whether you are scaling up a production strain or just starting your first fermentation run, this SOP ensures consistency, sterility, and operational safety.

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)

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

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