Standard Operating Procedure for Cleaning and Sanitization of Bioreactors in Biosimilar Manufacturing

Department	Biosimilars
SOP No.	SOP/BS/098/2025
Supersedes	SOP/BS/098/2022
Page No.	Page 1 of 12
Issue Date	04/05/2025
Effective Date	06/05/2025
Review Date	04/05/2026

1. Purpose

To define the validated cleaning and sanitization procedures for bioreactors used in biosimilar manufacturing, ensuring removal of residues and microbial control in accordance with cGMP standards.

2. Scope

This SOP applies to stainless-steel bioreactors and single-use bioreactor systems (SUBs) used in upstream processes of biosimilar production.

3. Responsibilities

• Manufacturing: Execute cleaning/sanitization as per validated protocols and document activities.
• Engineering: Maintain CIP/SIP systems, validate cleaning cycles, and troubleshoot equipment.
• QA: Approve cleaning validation reports and review cleaning logs.

4. Accountability

The Manufacturing Manager is accountable for ensuring bioreactors are cleaned and sanitized according to this SOP before and after each production batch.

5. Procedure

5.1 Cleaning of Stainless-Steel Bioreactors

1. Ensure the bioreactor is empty and all harvest lines are drained.
2. Initiate CIP cycle using the control panel:
   • Pre-rinse with WFI or RO water for 10 minutes.
   • Alkaline wash (e.g., 1% NaOH at 60°C for 20 minutes).
   • Intermediate rinse with WFI until conductivity <1 µS/cm.
   • Acid wash (e.g., 0.5% nitric acid) if required.
   • Final rinse with WFI.
3. Inspect internal surfaces using a borescope if accessible.

5.2 Sanitization (SIP) of Stainless-Steel Bioreactors

1. Perform SIP using clean steam (121°C) through all ports for minimum 30 minutes hold time.
2. Validate temperature probes reach ≥121°C during cycle.
3. Maintain log of time, temperature, and pressure (Annexure-2).

5.3 Cleaning and Sanitization of Single-Use Bioreactors

1. Use pre-sterilized bags with gamma irradiation certification.
2. If pre-use rinse required:
   • Rinse with sterile WFI for 5–10 minutes at ambient temperature.
3. Post-run, discard bag and sanitize shell with 70% IPA.

5.4 Cleaning Verification

1. Swab rinsing ports and interior for residual protein or cleaning agent.
2. Conduct TOC or conductivity test on rinse water; limits:
   • TOC < 500 ppb
   • Conductivity < 1.3 µS/cm

5.5 Documentation

1. Record all activities in Bioreactor Cleaning Log (Annexure-1).
2. Attach SIP printouts and cleaning validation results.

6. Abbreviations

• CIP: Clean-In-Place
• SIP: Sterilize-In-Place
• TOC: Total Organic Carbon
• QA: Quality Assurance

7. Documents

1. Bioreactor Cleaning Log – Annexure-1
2. SIP Cycle Monitoring Sheet – Annexure-2

8. References

• ICH Q9 – Quality Risk Management
• WHO TRS 1010 Annex 4 – GMP for Biological Products

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	Bioreactor ID	CIP Cycle Type	Operator	TOC Result	Remarks
04/05/2025	BR-SS-500L	Standard	Ajay Verma	410 ppb	Pass

Annexure-2: SIP Cycle Monitoring Sheet

Date	Cycle Start	Hold Start	Hold End	Min Temp (°C)	Operator
04/05/2025	09:00	09:10	09:40	122.3	Sunita Reddy

Revision History:

Revision Date	Revision No.	Revision Details	Reason for Revision	Approved By
04/05/2025	2.0	Included single-use bioreactor sanitization and TOC limits	Periodic GMP review