SOP Guide for Pharma

Biosimilars: SOP for Carbon Dioxide Sparging – V 2.0

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Biosimilars: SOP for Carbon Dioxide Sparging – V 2.0

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Standard Operating Procedure for Carbon Dioxide Sparging in Biosimilar Bioreactor Operations

Department Biosimilars
SOP No. SOP/BS/106/2025
Supersedes SOP/BS/106/2022
Page No. Page 1 of 11
Issue Date 04/05/2025
Effective Date 06/05/2025
Review Date 04/05/2026

1. Purpose

To define the procedure for carbon dioxide (CO₂) sparging in bioreactor systems during biosimilar upstream processes, primarily for pH control and maintaining an optimal environment for cell culture growth.

2. Scope

This SOP applies to all production-scale and pilot-scale bioreactor systems equipped with pH control modules that utilize CO₂ sparging as a corrective mechanism within the biosimilar manufacturing facility.

3. Responsibilities

  • Production Operator: Monitor and execute CO₂ sparging based on pH readings and process setpoints.
  • Supervisor: Approve manual sparging actions and review sparging logs.
  • Engineering: Maintain and calibrate flow meters and CO₂ delivery systems.
See also  Biosimilars: SOP for Hold Time Validation of Intermediate Products - V 2.0

4. Accountability

The Head of Production is accountable for ensuring the proper control and documentation of carbon dioxide sparging activities to maintain pH within acceptable process limits.

5. Procedure

5.1 System Setup and Pre-Check

  1. Verify CO₂ gas line is connected to the bioreactor gas mixing panel.
  2. Ensure CO₂ supply is medical grade or as per approved specification.
  3. Check leak-free connections and confirm functionality of backpressure regulators.

5.2 Setting pH Control Strategy

  1. Set desired pH control band (e.g., 7.0 ± 0.2) in the SCADA/control system.
  2. Define control cascade: Acid (CO₂) as primary downtrend mechanism, base (e.g., NaOH) as uptrend.

5.3 Initiating CO₂ Sparging

  1. If pH rises above upper setpoint, automated CO₂ sparging is triggered through MFC (Mass Flow Controller).
  2. For manual sparging:
    • Obtain supervisor approval.
    • Initiate flow through CO₂ inlet line at 0.01–0.05 vvm based on vessel volume.
  3. Monitor pH response continuously during enrichment.
See also  Biosimilars: SOP for Genetic Stability Testing - V 2.0

5.4 Monitoring and Documentation

  1. Log all CO₂ sparging events in Annexure-1 with time, duration, flow rate, and pH change.
  2. Ensure stable pH within control range is achieved within 15–30 minutes of sparging.

5.5 Post-Sparging Actions

  1. Close manual CO₂ flow valves and return control to automatic mode if overridden.
  2. Check for any foaming issues or DO shifts post-sparging.
  3. Notify Engineering if sparging is ineffective or erratic.

6. Abbreviations

  • CO₂: Carbon Dioxide
  • vvm: Volume of gas per volume of liquid per minute
  • MFC: Mass Flow Controller
  • SCADA: Supervisory Control and Data Acquisition

7. Documents

  1. CO₂ Sparging Log – Annexure-1
  2. CO₂ Flow Meter Calibration Record – Annexure-2
See also  Biosimilars: SOP for Training Bioreactor Operators - V 2.0

8. References

  • ICH Q10 – Pharmaceutical Quality System
  • FDA Guidance on Process Control and Monitoring (2023)

9. SOP Version

Version: 2.0

10. Approval Section

Prepared By Checked By Approved By
Signature
Date
Name
Designation
Department

11. Annexures

Annexure-1: CO₂ Sparging Log

Date Batch No. Initial pH Flow Rate (vvm) Start–End Time Final pH Performed By
04/05/2025 BS-215 7.3 0.03 10:10–10:30 7.0 Sunita Reddy

Annexure-2: CO₂ Flow Meter Calibration Record

Equipment ID Last Calibrated Due Date Status Calibrated By
MFC-CO2-204 01/04/2025 01/10/2025 OK Ajay Verma

Revision History:

Revision Date Revision No. Revision Details Reason for Revision Approved By
04/05/2025 2.0 Added flow rate limits, alarm response, and calibration annexure GMP process control update