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

1. Ensure agitation system is functional and calibrated (Annexure-1).
2. Inspect sparger and verify gas filters and valves are intact.
3. Set initial agitation speed (e.g., 80–120 RPM) as per batch record.
4. Configure aeration rate (typically 0.05–0.1 vvm) and oxygen blending setup.

5.2 Real-Time Monitoring

1. Record agitation speed and gas flow rate every 2 hours using the control system interface.
2. Monitor cascade control: agitation should respond to DO drops by ramping up gradually.
3. Check for foaming or vortex formation—adjust RPM if necessary.
4. Document readings in Annexure-2.

5.3 Adjustments During Culture

1. Increase agitation speed if DO or pH drifts despite gas adjustments.
2. Check for gas leaks or blockages if aeration is ineffective.
3. In case of foaming, refer to SOP/BS/072/2025 for antifoam addition.

5.4 Post-Run Handling

1. Shut down agitator and gas supply after harvesting.
2. Document shutdown parameters in Annexure-3.
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

1. Agitation & Aeration System Checklist – Annexure-1
2. Agitation & Aeration Monitoring Log – Annexure-2
3. 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