Standard Operating Procedure for Setup and Sterilization of Bioreactor Vessels in Biosimilar Production

Department	Biosimilars
SOP No.	SOP/BS/064/2025
Supersedes	SOP/BS/064/2022
Page No.	Page 1 of 12
Issue Date	04/05/2025
Effective Date	06/05/2025
Review Date	04/05/2026

1. Purpose

To establish a standardized procedure for assembly, cleaning, and sterilization of bioreactor vessels used in the upstream processing of biosimilars, ensuring aseptic operations and regulatory compliance.

2. Scope

This SOP is applicable to stainless steel and single-use bioreactors (S.U.B.) ranging from 1L to 2000L capacity used for mammalian cell culture in biosimilar manufacturing facilities.

3. Responsibilities

• Upstream Operators: Execute vessel setup and cleaning as per protocol.
• Maintenance Team: Calibrate sensors and validate autoclave/sterilizer functions.
• QA: Review sterilization and integrity records and approve for production use.

4. Accountability

The Upstream Production Head is accountable for ensuring bioreactor vessel integrity, aseptic readiness, and traceable sterilization records.

5. Procedure

5.1 Pre-Setup Verification

1. Ensure vessel has been properly cleaned after last use and all removable components are available (gaskets, agitator shaft, sampling ports, probes).
2. Review previous batch log for any deviations, contamination alerts, or maintenance flags.

5.2 Cleaning Procedure (Stainless Steel Bioreactor)

1. Perform visual inspection for residues.
2. Rinse vessel with purified water (PW).
3. Apply validated cleaning agent (e.g., 1% NaOH) and circulate using CIP (Clean-In-Place) system for 30 minutes.
4. Perform two rinse cycles with PW followed by one rinse with Water for Injection (WFI).
5. Dry using sterile filtered air and record in Cleaning Log (Annexure-1).

5.3 Assembly of Bioreactor

1. Install pH, DO, temperature, and pressure sensors ensuring correct orientation and calibration status.
2. Fit agitator, baffles, sparger, exhaust filters, and vent filters as per P&ID.
3. Secure connections with validated clamps and sterile tubing.

5.4 Sterilization Procedure

1. For SIP (Steam-In-Place):
   • Seal all ports and initiate automated SIP cycle (e.g., 121°C for 30 minutes).
   • Verify condensate drain and temperature mapping at multiple points.
2. For Autoclave Sterilization (Small-scale vessels):
   • Wrap removable components in sterile bags and autoclave at 121°C/15 psi for 30 minutes.

5.5 Post-Sterilization Verification

1. Perform filter integrity test (Bubble Point Test) for exhaust and vent filters (Annexure-2).
2. Record SIP validation parameters in Sterilization Log (Annexure-3).
3. Apply “Sterile Ready for Use” label post-QA release.

6. Abbreviations

• SIP: Steam-In-Place
• PW: Purified Water
• WFI: Water for Injection
• CIP: Clean-In-Place

7. Documents

1. Bioreactor Cleaning Log – Annexure-1
2. Filter Integrity Test Report – Annexure-2
3. SIP Sterilization Record – Annexure-3

8. References

• EU GMP Annex 1 – Sterile Manufacturing
• ICH Q7 – GMP for Active Pharmaceutical Ingredients
• WHO TRS 1010 – Biotech Production Guidelines

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name
Designation
Department

11. Annexures

Annexure-1: Bioreactor Cleaning Log

Date	Vessel ID	Cleaning Agent	Rinse Status	Operator
03/05/2025	SSV-500	1% NaOH	3x PW, 1x WFI	Ajay Verma

Annexure-2: Filter Integrity Test Report

Filter ID	Type	Test Method	Result	Tested By
F-EXP-01	PTFE	Bubble Point	Pass	QA Officer

Annexure-3: SIP Sterilization Record

Vessel ID	Start Time	Temperature	Hold Time	Validated By
SSV-500	10:00 AM	121°C	30 min	Sunita Reddy

Revision History:

Revision Date	Revision No.	Revision Details	Reason for Revision	Approved By
04/05/2025	2.0	Updated SIP validation steps and annexure formats	Process Audit Feedback