Standard Operating Procedure for Agitation and Aeration Monitoring in Bioreactor Operations for Biosimilars
| Department | Biosimilars |
|---|---|
| SOP No. | SOP/BS/071/2025 |
| Supersedes | SOP/BS/071/2022 |
| Page No. | Page 1 of 11 |
| Issue Date | 04/05/2025 |
| Effective Date | 06/05/2025 |
| Review Date | 04/05/2026 |
1. Purpose
To describe the procedure for monitoring and adjusting agitation and aeration parameters in bioreactor systems to ensure proper mixing, oxygen transfer, and cell viability during biosimilar production.
2. Scope
This SOP applies to all stirred-tank bioreactors (10L–2000L) used in upstream biosimilar processes involving mammalian cell culture systems such as CHO and HEK293 lines.
3. Responsibilities
- Process Operators: Monitor agitation RPM and gas flow rates, respond to alarms, and record data.
- Engineering: Maintain sparger, gas lines, and agitation motors.
- QA: Review logs and confirm adherence to specified process parameters.
4. Accountability
The Upstream Process Head is accountable for ensuring bioreactor agitation and aeration systems operate within validated parameters and deviations are appropriately handled.
5. Procedure
5.1 Pre-Run Checks
- Ensure agitation system is functional and calibrated (Annexure-1).
- Inspect sparger and verify gas filters and valves are intact.
- Set initial agitation speed (e.g., 80–120 RPM) as per batch record.
- Configure aeration rate (typically 0.05–0.1 vvm) and oxygen blending setup.
5.2 Real-Time Monitoring
- Record agitation speed and gas flow rate every 2 hours using the control system interface.
- Monitor cascade control: agitation should respond to DO drops by ramping up gradually.
- Check for foaming or vortex formation—adjust RPM if necessary.
- Document readings in Annexure-2.
5.3 Adjustments During Culture
- Increase agitation speed if DO or pH drifts despite gas adjustments.
- Check for gas leaks or blockages if aeration is ineffective.
- In case of foaming, refer to SOP/BS/072/2025 for antifoam addition.
5.4 Post-Run Handling
- Shut down agitator and gas supply after harvesting.
- Document shutdown parameters in Annexure-3.
- Schedule cleaning and maintenance of sparger and impeller.
6. Abbreviations
- RPM: Revolutions Per Minute
- vvm: Volume of gas per volume of liquid per minute
- DO: Dissolved Oxygen
7. Documents
- Agitation & Aeration System Checklist – Annexure-1
- Agitation & Aeration Monitoring Log – Annexure-2
- Shutdown Summary Report – Annexure-3
8. References
- WHO TRS 1010 – GMP for Biological Products
- ICH Q8 – Pharmaceutical Development
- SOP/BS/072/2025 – Antifoam Handling
9. SOP Version
Version: 2.0
10. Approval Section
| Prepared By | Checked By | Approved By | |
|---|---|---|---|
| Signature | |||
| Date | |||
| Name | |||
| Designation | |||
| Department |
11. Annexures
Annexure-1: Agitation & Aeration System Checklist
| Parameter | Status | Checked By | Date |
|---|---|---|---|
| Motor Functionality | OK | Rajesh Kumar | 04/05/2025 |
| Sparger Flow | OK | Rajesh Kumar | 04/05/2025 |
Annexure-2: Agitation & Aeration Monitoring Log
| Date | Time | RPM | Gas Flow (vvm) | Remarks |
|---|---|---|---|---|
| 04/05/2025 | 10:00 | 100 | 0.08 | Normal |
Annexure-3: Shutdown Summary Report
| Batch ID | Final RPM | Final Gas Flow | Shutdown Time | Operator |
|---|---|---|---|---|
| BR-113 | 80 | 0.00 | 19:15 | Ajay Verma |
Revision History:
| Revision Date | Revision No. | Revision Details | Reason for Revision | Approved By |
|---|---|---|---|---|
| 04/05/2025 | 2.0 | Added sparger inspection and Annexure-3 | Process Optimization |