Standard Operating Procedure for Microbial Contamination Check During Culture in Bioreactor Operations for Biosimilars

Department	Biosimilars
SOP No.	SOP/BS/079/2025
Supersedes	SOP/BS/079/2022
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 standard method for detecting microbial contamination during bioreactor operations in the biosimilar manufacturing process, ensuring timely identification and mitigation in compliance with GMP guidelines.

2. Scope

This SOP applies to all bioreactors (stainless steel and single-use) used for upstream biosimilar production that require routine microbial contamination checks during culture.

3. Responsibilities

• Process Operators: Collect in-process samples aseptically for microbial testing.
• Microbiology Analysts: Perform microbial testing, incubation, and reporting.
• QA Officer: Review reports and initiate CAPA in case of positive results.

4. Accountability

The Microbiology Manager is accountable for ensuring microbial contamination checks are conducted as per this SOP and any deviations are properly documented.

5. Procedure

5.1 Sampling Frequency and Materials

1. Collect samples once every 24 hours during culture, or as defined in the Batch Manufacturing Record (BMR).
2. Use sterile syringes or septum ports and sterile sample containers labeled with:
   • Batch No.
   • Date & Time
   • Bioreactor ID

5.2 Sample Collection Procedure

1. Wipe sampling port with 70% IPA and wear sterile gloves.
2. Withdraw 5–10 mL of culture fluid using sterile technique.
3. Transfer immediately to labeled container and send to Microbiology Lab within 30 minutes.
4. Log collection details in Annexure-1.

5.3 Microbiological Testing

1. Perform the following tests:
   • Total Aerobic Count (TAMC)
   • Total Yeast and Mold Count (TYMC)
   • Gram Staining
   • Rapid Mycoplasma Detection (if needed)
2. Inoculate 1 mL sample into TSB and FTM media and incubate at 20–25°C and 30–35°C respectively for 7 days.
3. Record test results daily for contamination signs (Annexure-2).

5.4 Result Interpretation and Actions

1. If growth is observed, immediately:
   • Quarantine the bioreactor
   • Collect second confirmatory sample
   • Inform QA and initiate deviation report
2. If confirmed, terminate batch and initiate CAPA and cleaning validation.
3. Document all results and corrective measures in Annexure-3.

6. Abbreviations

• TAMC: Total Aerobic Microbial Count
• TYMC: Total Yeast and Mold Count
• CAPA: Corrective and Preventive Action
• IPA: Isopropyl Alcohol

7. Documents

1. Sampling Log – Annexure-1
2. Microbial Test Observation Log – Annexure-2
3. Deviation and CAPA Record – Annexure-3

8. References

• USP <61> and <62> – Microbiological Examination
• WHO TRS 1004 – GMP for Biotech Products
• ICH Q9 – Quality Risk 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: Sampling Log

Date	Time	Batch No.	Reactor ID	Collected By	Remarks
04/05/2025	09:00	BS-079-B01	BR-03	Rajesh Kumar	Normal

Annexure-2: Microbial Test Observation Log

Date	Media Type	Incubation Temp	Observation	Analyst
04/05/2025	TSB	30–35°C	No Growth	Sunita Reddy

Annexure-3: Deviation and CAPA Record

Date	Deviation Description	Immediate Action	CAPA	QA Review
04/05/2025	Positive Bacterial Growth	Batch Hold	Re-training & Sanitation	Pending

Revision History:

Revision Date	Revision No.	Revision Details	Reason for Revision	Approved By
04/05/2025	2.0	Added rapid detection method and Annexure-3 CAPA record	Compliance Upgrade