Standard Operating Procedure for Scale-Up from Shake Flask to Spinner Flask in Biosimilar Production
| Department | Biosimilars |
|---|---|
| SOP No. | SOP/BS/066/2025 |
| Supersedes | SOP/BS/066/2022 |
| 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 aseptic scale-up of seed cultures from shake flasks to spinner flasks as part of the biosimilar upstream process, maintaining cell viability, sterility, and traceability in accordance with GMP.
2. Scope
This SOP applies to all scale-up operations from shake flasks (125 mL–1 L) to spinner flasks (2 L–5 L) used in biosimilar manufacturing for CHO, HEK293, and other mammalian cell lines.
3. Responsibilities
- Upstream Technician: Perform inoculation and maintain spinner flask logs.
- Process Scientist: Monitor growth parameters and determine scale-up readiness.
- QA Personnel: Review batch documentation and ensure environmental compliance.
4. Accountability
The Upstream Process Manager is accountable for seed culture integrity and ensuring that scale-up activities meet GMP and process development standards.
5. Procedure
5.1 Readiness Assessment
- Confirm shake flask culture has VCD ≥ 1.0 × 10⁶ cells/mL and viability ≥ 90%.
- Document current culture health in Annexure-1.
5.2 Spinner Flask Setup
- Retrieve pre-sterilized spinner flasks (2 L–5 L) with magnetic or paddle stirrers.
- Label flask with batch number, media type, date, and flask ID (Annexure-2).
- Transfer sterile chemically defined media (CDM) to spinner flask under LAF.
5.3 Aseptic Inoculation
- Gently mix shake flask culture and withdraw required volume using sterile pipettes.
- Inoculate spinner flask to achieve VCD of ~0.3 × 10⁶ cells/mL.
- Secure flask lid and place on magnetic or mechanical stirrer (45–60 rpm) in incubator shaker or CO₂ incubator at 37°C, 5% CO₂.
5.4 Monitoring and Control
- Record daily VCD, viability, pH, temperature, and osmolality (Annexure-3).
- Perform sterility checks on Day 0 and Day 3 per SOP/BS/043/2025.
- Inspect visually for turbidity, sediment, or abnormal morphology.
5.5 Documentation and Review
- Update seed train map and transfer batch record to process scientist for review.
- QA to verify Annexure entries and approve for bioreactor inoculation.
6. Abbreviations
- VCD: Viable Cell Density
- CDM: Chemically Defined Media
- LAF: Laminar Air Flow
- GMP: Good Manufacturing Practice
7. Documents
- Shake Flask Culture Assessment Sheet – Annexure-1
- Spinner Flask Label Log – Annexure-2
- Daily Monitoring Log – Annexure-3
8. References
- ICH Q5D – Cell Substrates for Biotechnological/Biological Products
- WHO TRS 1010 – Bioprocess Guidelines
- SOP/BS/043/2025 – Sterility Monitoring
9. SOP Version
Version: 2.0
10. Approval Section
| Prepared By | Checked By | Approved By | |
|---|---|---|---|
| Signature | |||
| Date | |||
| Name | |||
| Designation | |||
| Department |
11. Annexures
Annexure-1: Shake Flask Culture Assessment Sheet
| Date | Flask ID | VCD (×10⁶/mL) | Viability (%) | Approved for Scale-Up |
|---|---|---|---|---|
| 03/05/2025 | FLK-005 | 1.2 | 92 | Yes |
Annexure-2: Spinner Flask Label Log
| Flask ID | Volume (L) | Media Type | Date of Inoculation | Operator |
|---|---|---|---|---|
| SPF-01 | 2 | CD CHO | 04/05/2025 | Ajay Verma |
Annexure-3: Daily Monitoring Log
| Date | VCD | Viability | pH | Remarks |
|---|---|---|---|---|
| 05/05/2025 | 0.85 | 94% | 7.1 | Healthy Growth |
Revision History:
| Revision Date | Revision No. | Revision Details | Reason for Revision | Approved By |
|---|---|---|---|---|
| 04/05/2025 | 2.0 | Included osmolality check and new annexures | GMP Audit Requirement |