Standard Operating Procedure for Creation of Master Cell Bank (MCB) in Biosimilars
| Department | Biosimilars |
|---|---|
| SOP No. | SOP/BS/024/2025 |
| Supersedes | SOP/BS/024/2022 |
| Page No. | Page 1 of 14 |
| Issue Date | 04/05/2025 |
| Effective Date | 06/05/2025 |
| Review Date | 04/05/2026 |
1. Purpose
To outline the procedure for the preparation, qualification, documentation, and storage of Master Cell Banks (MCBs) derived from biosimilar-producing cell lines intended for GMP manufacturing.
2. Scope
This SOP applies to the Cell Line Development and Quality Control departments involved in the generation, testing, and archival of MCBs under GMP conditions.
3. Responsibilities
- CLD Scientist: Prepares and freezes MCB vials under controlled aseptic conditions.
- QC Microbiologist: Conducts sterility, mycoplasma, identity, and viability testing.
- QA Officer: Ensures all MCB-related activities are documented, reviewed, and approved as per GMP standards.
4. Accountability
The Head of Biosimilars is accountable for ensuring that the MCB is prepared, characterized, and stored in compliance with regulatory expectations and internal quality systems.
5. Procedure
5.1 Pre-Freezing Preparation
- Use a genetically and phenotypically stable cell clone that has passed clone stability studies (refer SOP/BS/023/2025).
- Thaw and expand the selected clone in a progressive scale-up from T-flasks → spinner flasks → production bioreactor (if applicable).
- Monitor viable cell density (>90% viability) before harvest.
5.2 Cryopreservation Media
- Prepare freezing medium: 90% basal media + 10% DMSO under sterile conditions.
- Filter sterilize the medium using 0.22 µm filters.
5.3 MCB Aliquoting and Labeling
- Resuspend cells at 5 × 106 cells/mL in freezing media.
- Dispense 1 mL into pre-labeled sterile cryovials (minimum 100 vials recommended).
- Labels must include:
- Cell Line ID
- MCB Lot Number
- Date of Freezing
- Passage Number
- Technician Initials
5.4 Controlled Freezing
- Place vials into a programmable rate freezer: -1°C/min until -80°C.
- Transfer to liquid nitrogen (-196°C) storage tank after 12–24 hours.
5.5 Quality Control Testing
- Send representative vials for:
- Cell viability
- Sterility (as per USP <71>)
- Mycoplasma (culture and PCR-based)
- Cell line identity (STR profiling)
- Adventitious virus testing (where required)
- Record all results in MCB QC Log (Annexure-1).
5.6 Documentation and Archival
- Maintain freezing records and storage inventory in MCB Inventory Log (Annexure-2).
- Prepare MCB Certificate of Analysis upon completion of QC testing.
6. Abbreviations
- MCB: Master Cell Bank
- DMSO: Dimethyl Sulfoxide
- STR: Short Tandem Repeat
- QC: Quality Control
7. Documents
- MCB QC Log (Annexure-1)
- MCB Inventory Log (Annexure-2)
8. References
- ICH Q5D – Derivation and Characterization of Cell Substrates
- WHO Technical Report Series No. 978 – Cell Substrate Guidelines
- CDSCO Biosimilar Guidelines (India)
9. SOP Version
Version: 2.0
10. Approval Section
| Prepared By | Checked By | Approved By | |
|---|---|---|---|
| Signature | |||
| Date | |||
| Name | |||
| Designation | |||
| Department |
11. Annexures
Annexure-1: MCB QC Log
| Test | Method | Result | Acceptance Criteria | Remarks |
|---|---|---|---|---|
| Viability | Trypan Blue | 94% | ≥ 85% | Pass |
| Sterility | USP <71> | Negative | No growth | Pass |
| Mycoplasma | PCR + Culture | Negative | Absent | Pass |
| STR Profile | Capillary Electrophoresis | Matched | 100% match | Pass |
Annexure-2: MCB Inventory Log
| Vial No. | Cell Line ID | MCB Lot No. | Freezing Date | Location (Tank/Rack/Box) | Operator |
|---|---|---|---|---|---|
| MCB001 | CL-BS-024 | MCB-BS-24A | 03/05/2025 | T1/R2/B4 | Sunita Reddy |
Revision History:
| Revision Date | Revision No. | Revision Details | Reason for Revision | Approved By |
|---|---|---|---|---|
| 04/05/2025 | 2.0 | Updated storage practices and included STR and virus testing | Regulatory update and audit recommendation |