Standard Operating Procedure for Seed Train Validation in Biosimilar Manufacturing
| Department | Biosimilars |
|---|---|
| SOP No. | SOP/BS/107/2025 |
| Supersedes | SOP/BS/107/2022 |
| 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 procedure for validation of the seed train process used in upstream biosimilar production, ensuring consistency of cell growth and quality attributes from thaw to production-scale inoculum.
2. Scope
This SOP applies to all stages of the seed train from vial thawing, shake flask, spinner flask, and seed bioreactor expansions up to the point of inoculation into the main production bioreactor for biosimilar manufacturing.
3. Responsibilities
- Process Development: Define seed train parameters and validation protocol.
- Manufacturing: Execute seed train stages and record all critical data.
- QA: Review validation data, approve protocol and final report.
4. Accountability
The Head of Manufacturing Sciences is accountable for ensuring that seed train validation is performed, approved, and maintained in compliance with regulatory expectations.
5. Procedure
5.1 Preparation and Planning
- Prepare Seed Train Validation Protocol including:
- Acceptance criteria for growth, viability, and cell density
- Scalability criteria (VCD growth factor, doubling time)
- Sampling points and analytical methods
- Obtain QA approval before execution.
5.2 Seed Train Execution
- Start with a thawed vial from a qualified Master Cell Bank.
- Expand through sequential vessels:
- Shake flask (50–250 mL)
- Spinner flask or Wave bag (500 mL–2 L)
- Seed bioreactor (5 L–200 L)
- Monitor and record:
- Viable Cell Density (VCD)
- Cell viability
- pH, temperature, DO
- Contamination (visual and microbial)
5.3 Sampling and Testing
- Collect samples at each expansion stage:
- Start and end of each vessel
- Additional time points as required
- Analyze for:
- Growth rate and doubling time
- Glucose consumption, lactate production
- Viability ≥ 90% at end of each stage
5.4 Data Analysis and Documentation
- Document all results in Annexure-1 and Annexure-2.
- Evaluate data against pre-defined criteria.
- Summarize in Seed Train Validation Report and submit to QA.
5.5 Revalidation Criteria
- Revalidate seed train upon:
- Change in media or supplements
- Change in vessel configuration or scale
- New cell bank introduction
6. Abbreviations
- VCD: Viable Cell Density
- QA: Quality Assurance
- MCC: Master Cell Bank
- DSP: Downstream Processing
7. Documents
- Seed Train Execution Log – Annexure-1
- Seed Train Validation Checklist – Annexure-2
8. References
- ICH Q8 – Pharmaceutical Development
- 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: Seed Train Execution Log
| Date | Stage | VCD (10⁶ cells/mL) | Viability (%) | Glucose (g/L) | Remarks |
|---|---|---|---|---|---|
| 04/05/2025 | Shake Flask | 1.2 | 94 | 2.5 | Normal |
Annexure-2: Seed Train Validation Checklist
| Parameter | Target | Observed | Status | Reviewed By |
|---|---|---|---|---|
| Doubling Time | 18–24 hrs | 21 hrs | Pass | Ajay Verma |
Revision History:
| Revision Date | Revision No. | Revision Details | Reason for Revision | Approved By |
|---|---|---|---|---|
| 04/05/2025 | 2.0 | Updated validation metrics and added revalidation triggers | Process harmonization |