Standard Operating Procedure for Inactivation of Used Culture Broth in Biosimilar Manufacturing

Department	Biosimilars
SOP No.	SOP/BS/113/2025
Supersedes	SOP/BS/113/2022
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 safe, compliant, and effective inactivation of used culture broth from biosimilar upstream processes to prevent environmental contamination and ensure biosafety before disposal.

2. Scope

This SOP applies to all spent culture broths and residual biomass generated during biosimilar cell culture operations in seed bioreactors, production bioreactors, and media holding tanks within the upstream manufacturing facility.

3. Responsibilities

• Production Operator: Execute broth inactivation using approved methods and document activity.
• Utility/Engineering Team: Ensure availability of heating or chemical dosing systems.
• QA: Review inactivation records and verify compliance.

4. Accountability

The Head of Production is accountable for ensuring all used culture broths are rendered non-infectious and disposed of as per regulatory standards.

5. Procedure

5.1 Pre-Inactivation Checks

1. Confirm culture harvest completion and authorization for inactivation.
2. Verify inactivation vessel is cleaned, sanitized, and calibrated.
3. Record batch number, volume of culture broth, and vessel ID in Annexure-1.

5.2 Inactivation Methods

5.2.1 Thermal Inactivation

1. Ensure vessel jacket or coil heating system is functional.
2. Set target temperature: 121°C for 30 minutes.
3. Start heating cycle and monitor via temperature gauge or SCADA system.
4. Record start and end time of hold period and confirm temperature profile.

5.2.2 Chemical Inactivation (Alternative)

1. Prepare 1% sodium hypochlorite or validated chemical disinfectant.
2. Add to vessel at 10% v/v under agitation.
3. Hold for minimum 1 hour ensuring contact with all surfaces and broth mass.
4. Confirm pH (10–11) and perform microbial kill test if applicable.

5.3 Post-Inactivation Confirmation

1. Record final parameters in Annexure-2.
2. Take sample (optional) and send to QC for total viable count (TVC) testing.
3. Label vessel or transfer line as “Inactivated” post-confirmation.

5.4 Disposal

1. Drain inactivated broth into Effluent Treatment Plant (ETP) or through designated deactivation pit.
2. Flush lines with WFI or potable water as per standard cleaning protocol.

6. Abbreviations

• SCADA: Supervisory Control and Data Acquisition
• ETP: Effluent Treatment Plant
• TVC: Total Viable Count
• QA: Quality Assurance

7. Documents

1. Broth Inactivation Log – Annexure-1
2. Inactivation Confirmation Sheet – Annexure-2

8. References

• WHO TRS 999 – Environmental Biosafety in Biopharmaceutical Facilities
• EPA Guidelines for Biohazardous Waste Management

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name
Designation
Department

11. Annexures

Annexure-1: Broth Inactivation Log

Date	Batch No.	Volume (L)	Method	Operator	Remarks
04/05/2025	BS-BT-301	200	Thermal	Sunita Reddy	Cycle OK

Annexure-2: Inactivation Confirmation Sheet

Parameter	Set Value	Achieved	Hold Time	Verified By
Temperature	121°C	121.4°C	30 min	Ajay Verma

Revision History:

Revision Date	Revision No.	Revision Details	Reason for Revision	Approved By
04/05/2025	2.0	Added chemical inactivation as alternate method and ETP reference	Process enhancement