Standard Operating Procedure for In-Process Sampling Procedures in Bioreactor Operations for Biosimilars

Department	Biosimilars
SOP No.	SOP/BS/078/2025
Supersedes	SOP/BS/078/2022
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 standardized method for aseptic in-process sampling from bioreactors used in biosimilar production. This SOP ensures representative, contaminant-free samples for analysis of critical process parameters.

2. Scope

This SOP applies to all stainless steel and single-use bioreactor systems used in upstream biosimilar manufacturing for sampling culture media, cells, metabolites, and gases.

3. Responsibilities

• Bioprocess Technicians: Perform in-process sampling and record data.
• QC Analysts: Analyze collected samples for cell count, viability, pH, metabolites, etc.
• QA Officer: Verify logbook entries and confirm sampling frequencies.

4. Accountability

The Upstream Production Manager is accountable for ensuring in-process sampling is performed aseptically and results are promptly reviewed.

5. Procedure

5.1 Sampling Frequency

1. Follow the sampling schedule as per the approved Batch Manufacturing Record (BMR).
2. Typical sampling intervals:
   • Every 12 hours during the batch phase
   • Every 8 hours during the feeding phase
   • Every 4 hours near harvest phase

5.2 Preparation

1. Ensure all sampling accessories are sterile (syringes, sample bags, tubes).
2. Wipe sampling ports with 70% IPA and ensure sterile gloves and gowning.
3. Label sample containers with:
   • Batch No.
   • Date and Time
   • Sample Type

5.3 Sampling Technique

1. Connect sampling line or septum port to a sterile syringe or sampling valve.
2. Discard the first 2–5 mL to remove stagnant fluid (flush step).
3. Collect the required volume (typically 10–20 mL) without introducing air bubbles.
4. Seal sample container immediately and transfer to QC or store at 2–8°C until analysis.

5.4 Post Sampling

1. Disinfect sampling port again with 70% IPA and close securely.
2. Dispose of used syringes and gloves as per biohazard SOPs.
3. Record the sampling details in the Bioreactor Sampling Log (Annexure-1).

5.5 Sample Handling and Transfer

1. Transport samples in designated clean containers to QC within 30 minutes of collection.
2. Log handover with time and recipient signature in Sample Transfer Log (Annexure-2).

6. Abbreviations

• BMR: Batch Manufacturing Record
• QC: Quality Control
• IPA: Isopropyl Alcohol

7. Documents

1. Bioreactor Sampling Log – Annexure-1
2. Sample Transfer Log – Annexure-2

8. References

• ICH Q9 – Quality Risk Management
• WHO TRS 999 – 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 Sampling Log

Date	Time	Batch No.	Sample Type	Volume (mL)	Collected By	Remarks
04/05/2025	08:00	BS-BR-078	Viability	15	Rajesh Kumar	Normal

Annexure-2: Sample Transfer Log

Date	Time	Sample ID	Transferred To	Signature
04/05/2025	08:15	BS-078-08V	Sunita Reddy	SR

Revision History:

Revision Date	Revision No.	Revision Details	Reason for Revision	Approved By
04/05/2025	2.0	Updated sample handling and transfer timelines	GMP Compliance