Standard Operating Procedure for Centrifuge Operation for Cell Removal in Biosimilar Manufacturing

Department	Biosimilars
SOP No.	SOP/BS/152/2025
Supersedes	SOP/BS/152/2022
Page No.	Page 1 of 10
Issue Date	04/05/2025
Effective Date	06/05/2025
Review Date	04/05/2026

1. Purpose

To provide a detailed procedure for the operation of centrifuges for removing cells and cell debris from harvested biosimilar culture fluid prior to downstream processing under Good Manufacturing Practice (GMP) guidelines.

2. Scope

This SOP is applicable to the use of continuous-flow or batch centrifuges used in biosimilar upstream processing facilities for the separation of biomass post-harvest.

3. Responsibilities

• Production: Set up, operate, and monitor centrifuge system; document operations and report deviations.
• Engineering: Perform equipment maintenance and calibration as per schedule.
• QA: Review logbooks and approve operation before and after use for GMP batches.

4. Accountability

The Upstream Manufacturing Lead is accountable for ensuring compliance with the centrifuge operation procedure during batch harvest in biosimilar production.

5. Procedure

5.1 Pre-Operational Checks

1. Verify equipment ID, status label, and cleaning status.
2. Ensure balance of rotor assembly and inspect seals and gaskets.
3. Check calibration validity of RPM, temperature, and timer controls.
4. Record pre-checks in Annexure-1: Centrifuge Pre-Use Checklist.

5.2 Loading and Setup

1. Connect the harvest vessel outlet to the centrifuge inlet line using sterile connectors.
2. Ensure the clarified supernatant outlet and sludge outlet lines are properly routed to respective collection tanks.
3. Set desired parameters on the control panel:
   • RPM: 3500–6000 (as per process)
   • Feed flow rate: 1–3 L/min
   • Discharge cycle: As defined in batch record

5.3 Operation

1. Start the centrifuge and allow it to reach set RPM.
2. Begin feeding harvest fluid at low flow and gradually increase to setpoint.
3. Monitor:
   • Motor load
   • Vibration levels
   • Feed tank level
   • Clarified output turbidity (if applicable)
4. Periodically discharge sludge manually or automatically as per cycle setting.

5.4 Completion and Cleaning

1. Once feed is complete, run rinse cycle with WFI or designated buffer for 5–10 minutes.
2. Shut down centrifuge following OEM procedure.
3. Disassemble rotors and clean surfaces if required as per Annexure-2: Cleaning Log.

5.5 Documentation

1. Enter operation data including:
   • Start/end time
   • Total volume processed
   • RPM and cycle durations
   • Observations or abnormalities
2. Attach printouts from control panel (if available) to batch documentation.

5.6 Deviation Management

1. Report any of the following as deviation:
   • RPM deviation beyond 5%
   • Leakage or foam formation
   • Unexpected shutdown or discharge failure
2. Record in deviation log and inform QA immediately.

6. Abbreviations

• RPM: Revolutions Per Minute
• WFI: Water for Injection
• OEM: Original Equipment Manufacturer
• QA: Quality Assurance

7. Documents

1. Centrifuge Pre-Use Checklist – Annexure-1
2. Centrifuge Cleaning Log – Annexure-2
3. Centrifuge Operation Summary – Annexure-3

8. References

• WHO TRS 999 – GMP for Biotherapeutic Products
• OEM Equipment Manual – Alfa Laval / GE Centritech
• EU GMP Annex 1 – Manufacture of Sterile Medicinal 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: Centrifuge Pre-Use Checklist

Date	Equipment ID	Rotor Check	Seals Check	Status	Checked By
04/05/2025	CF-102	OK	OK	Ready	Ajay Verma

Annexure-2: Centrifuge Cleaning Log

Date	Equipment ID	Cleaning Agent	Contact Time	Rinsed (Y/N)	Cleaned By
04/05/2025	CF-102	NaOH 0.5%	10 min	Y	Sunita Reddy

Annexure-3: Centrifuge Operation Summary

Batch No.	Start Time	End Time	RPM	Volume (L)	Operator
BS-DP-081	09:00	10:15	4200	85	Sunita Reddy

Revision History:

Revision Date	Revision No.	Revision Details	Reason for Revision	Approved By
04/05/2025	2.0	Included clarification on flow rate and cleaning validation	Process standardization