Standard Operating Procedure for Use of Single-Use Bioreactor Systems in Biosimilar Manufacturing

Department	Biosimilars
SOP No.	SOP/BS/100/2025
Supersedes	SOP/BS/100/2022
Page No.	Page 1 of 13
Issue Date	04/05/2025
Effective Date	06/05/2025
Review Date	04/05/2026

1. Purpose

To define the standard procedure for the preparation, operation, and disposal of single-use bioreactor (SUB) systems used in biosimilar manufacturing, ensuring aseptic processing and operational consistency.

2. Scope

This SOP applies to all single-use bioreactors ranging from 50L to 2000L capacity used in upstream processing within the biosimilars facility.

3. Responsibilities

• Production: Set up, monitor, and dispose of SUB systems per batch requirement.
• Engineering: Verify equipment integrity, calibrate load cells, and support automation interfaces.
• QA: Ensure SUB components are qualified and that batch documentation complies with GMP standards.

4. Accountability

The Manufacturing Head is accountable for ensuring proper utilization and documentation of SUB systems as per validated procedures.

5. Procedure

5.1 Receipt and Inspection of SUB Components

1. Receive pre-sterilized bioreactor bag assemblies with gamma irradiation certification (Annexure-1).
2. Verify integrity of ports, sensors, weld lines, and bag labeling.
3. Check irradiation indicator color change and record batch number.

5.2 Assembly and Installation

1. Install the bag into the SUB vessel and connect sparger, dip tube, pH/DO probes, and sampling lines.
2. Use aseptic connections (e.g., AseptiQuik) for media, feed, and vent lines.
3. Secure probes and tighten clamps to prevent leaks or disconnections.

5.3 System Integrity Testing

1. Conduct leak test by filling bag with 10–20 L sterile air or nitrogen and applying pressure of 0.2 bar.
2. Monitor for 10 minutes and confirm no pressure drop.
3. Record test in Annexure-2: SUB Integrity Log.

5.4 Media Charging and Inoculation

1. Transfer sterile media through aseptic transfer line using peristaltic pump.
2. Confirm temperature, pH, and DO sensors are calibrated and functional.
3. Inoculate culture using sterile connections and begin agitation and aeration based on protocol.

5.5 Operation Monitoring

1. Monitor bioreactor parameters via control system (e.g., DeltaV, BioPAT).
2. Record agitation speed, pH, DO, VCD, and temperature every 4 hours or as per BMR.
3. Document all interventions in Annexure-3: Bioreactor Operation Log.

5.6 Harvest and Disposal

1. Transfer culture broth to harvest tank via sterile connections.
2. Drain and seal used SUB bag in biohazard bag and place in designated disposal bin.
3. Sanitize SUB vessel with 70% IPA before storage.

6. Abbreviations

• SUB: Single-Use Bioreactor
• BMR: Batch Manufacturing Record
• IPA: Isopropyl Alcohol
• QA: Quality Assurance

7. Documents

1. Gamma Irradiation Certificate – Annexure-1
2. SUB Integrity Log – Annexure-2
3. Bioreactor Operation Log – Annexure-3

8. References

• FDA Guidance for Industry – Single-Use Technology in Biopharmaceutical Manufacturing (2021)
• 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: Gamma Irradiation Certificate

Bag ID	Lot No.	Irradiation Date	Indicator Color	Checked By
SUB-100L-21	GIR-789	01/05/2025	Orange to Red	Ajay Verma

Annexure-2: SUB Integrity Log

Date	SUB ID	Test Pressure (bar)	Hold Time	Result	Performed By
04/05/2025	SUB-200L-35	0.2	10 min	Pass	Rajesh Kumar

Annexure-3: Bioreactor Operation Log

Time	pH	DO (%)	Temperature (°C)	Agitation (rpm)	Operator
08:00	7.1	40	37.0	120	Sunita Reddy

Revision History:

Revision Date	Revision No.	Revision Details	Reason for Revision	Approved By
04/05/2025	2.0	Included leak test procedure and annexures for irradiation certificate and logs	Periodic review and alignment with FDA guidelines