Standard Operating Procedure for Centrifugation of Harvested Broth in Bioreactor Operations for Biosimilars

Department	Biosimilars
SOP No.	SOP/BS/087/2025
Supersedes	SOP/BS/087/2022
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 procedure for centrifuging harvested culture broth from bioreactor operations during biosimilar manufacturing, with the aim of separating biomass and collecting clarified supernatant for downstream processing.

2. Scope

This SOP applies to all large-scale bioreactor harvests from mammalian or microbial cultures that require cell removal through centrifugation prior to filtration or purification.

3. Responsibilities

• Production Operator: Execute centrifugation as per batch instructions and monitor run parameters.
• Maintenance Engineer: Ensure centrifuge calibration, balancing, and operational readiness.
• QA Officer: Review centrifuge logs and ensure compliance with GMP documentation.

4. Accountability

The Production Head is accountable for verifying that centrifugation is performed as per defined parameters and ensuring safe operation of equipment and transfer of supernatant.

5. Procedure

5.1 Equipment Preparation

1. Verify cleanliness status of centrifuge and attach “Ready for Use” status tag.
2. Check rotor integrity, seals, and absence of residue or contamination.
3. Ensure that collection containers and transfer lines are sanitized and labeled with batch details.

5.2 Loading Harvested Broth

1. Connect feed line from harvest tank to centrifuge inlet under aseptic conditions.
2. Prime lines using 0.9% saline or WFI (if applicable).
3. Ensure vessel volume and loading balance as per capacity guidelines (e.g., max 400 L per cycle for swing-bucket rotor).

5.3 Centrifugation Parameters

1. Set speed: 4000–6000 rpm (depending on cell type and centrifuge model).
2. Run time: 10–20 minutes per cycle.
3. Set temperature: 4°C for mammalian cells, ambient for microbial cultures.
4. Program automatic braking and acceleration curves to minimize cell disruption.

5.4 Supernatant Collection

1. After each run, transfer clear supernatant to the clarified harvest tank using sterile connection.
2. Label collection tank with:
   • Product name
   • Batch number
   • Date and time of harvest
3. Take a 10 mL sample and send to QC for turbidity and bioburden check.

5.5 Post-Centrifugation Activities

1. Flush the centrifuge bowl and rotor with WFI and inspect for residual cells.
2. Record all operational parameters in Annexure-1: Centrifugation Log.
3. Initiate cleaning SOP post-harvest cycle or before product changeover.

6. Abbreviations

• rpm: Revolutions Per Minute
• QC: Quality Control
• WFI: Water for Injection
• GMP: Good Manufacturing Practice

7. Documents

1. Centrifugation Log – Annexure-1

8. References

• ICH Q7 – Good Manufacturing Practice Guide for Active Pharmaceutical Ingredients
• WHO TRS 1025 Annex 2 – GMP for Biotech 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: Centrifugation Log

Date	Batch No.	Cycle No.	Volume (L)	Speed (rpm)	Time (min)	Temperature (°C)	Operator
04/05/2025	BS-087-B02	1	400	5000	15	4	Sunita Reddy

Revision History:

Revision Date	Revision No.	Revision Details	Reason for Revision	Approved By
04/05/2025	2.0	Added specific centrifuge settings and bioburden sample collection	Process Clarification