Standard Operating Procedure for Selecting Host Cell Lines in Biosimilar Manufacturing
| Department | Biosimilars |
|---|---|
| SOP No. | SOP/BS/001/2025 |
| Supersedes | SOP/BS/001/2022 |
| Page No. | Page 1 of 12 |
| Issue Date | 04/05/2025 |
| Effective Date | 06/05/2025 |
| Review Date | 04/05/2026 |
1. Purpose
To establish a standardized procedure for selecting host cell lines such as CHO, HEK293, and E. coli for the development of biosimilars, ensuring high productivity, regulatory compliance, and reproducibility.
2. Scope
This SOP applies to all personnel in the Biosimilars department involved in the selection, evaluation, and documentation of host cell lines during early-stage biosimilar development.
3. Responsibilities
- Research Scientists: Perform literature review and technical assessments for host cell line suitability.
- Process Development Team: Conduct experimental validation of expression and growth performance.
- Regulatory Affairs (RA): Ensure the selected cell lines meet global regulatory and biosafety standards.
- Quality Assurance (QA): Review and archive all documentation related to cell line selection.
4. Accountability
The Head of Biosimilars is accountable for ensuring compliance with this SOP and the scientific and regulatory appropriateness of the host cell lines selected.
5. Procedure
5.1 Define Product Requirements
- Identify the biosimilar product type: e.g., monoclonal antibody, cytokine, or hormone.
- Define requirements for post-translational modifications such as glycosylation.
- Establish expression yield targets based on commercial-scale requirements.
5.2 Compile List of Candidate Host Cell Lines
- Generate a list of commonly used host cell lines:
- CHO (Chinese Hamster Ovary)
- HEK293 (Human Embryonic Kidney)
- E. coli (for prokaryotic expression)
- Assess each host cell line based on:
- Scalability
- Doubling time and metabolic efficiency
- Post-translational modification capabilities
- Availability of safety and lineage documentation
5.3 Perform Feasibility Experiments
- Perform small-scale transfection experiments with model constructs.
- Measure protein expression levels using ELISA or SDS-PAGE.
- Record findings in the Host Cell Screening Log (Annexure-1).
5.4 Review Regulatory and Biosafety Compliance
- Check biosafety levels (BSL-1 or BSL-2) of selected cell lines.
- Ensure the availability of cell bank Certificate of Analysis (CoA).
- Verify compliance with ICH Q5D, WHO TRS 978, and CDSCO requirements.
5.5 Final Selection and Approval
- Consolidate all experimental, regulatory, and biosafety data.
- Complete the Host Cell Line Selection Form (Annexure-2).
- Submit the form for cross-functional review and approval.
5.6 Documentation
- Update the Host Cell Line Master List (Annexure-3).
- File all relevant data in the Cell Line Selection Master File.
6. Abbreviations
- SOP: Standard Operating Procedure
- CHO: Chinese Hamster Ovary
- HEK293: Human Embryonic Kidney 293
- CoA: Certificate of Analysis
- RA: Regulatory Affairs
- QA: Quality Assurance
7. Documents
- Host Cell Screening Log (Annexure-1)
- Host Cell Line Selection Form (Annexure-2)
- Host Cell Line Master List (Annexure-3)
8. References
- ICH Q5D: Cell Substrate Derivation and Characterization
- WHO TRS 978 Annex 3
- CDSCO Guidelines – Cell Line Selection (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: Host Cell Screening Log
| Date | Cell Line | Transfection Method | Expression Yield | Remarks |
|---|---|---|---|---|
| 22/04/2025 | CHO-K1 | Electroporation | Medium | Good viability |
| 23/04/2025 | HEK293 | PEI | High | Selected |
| 24/04/2025 | E. coli DH5α | Heat Shock | Very High | No glycosylation |
Annexure-2: Host Cell Line Selection Form
| Selected Cell Line | HEK293 |
|---|---|
| Justification | High expression yield and compatible glycosylation |
| Date of Selection | 26/04/2025 |
| Cross-Functional Review Team | QA, RA, R&D |
Annexure-3: Host Cell Line Master List
| Cell Line | Type | BSL | Source | CoA No. |
|---|---|---|---|---|
| HEK293 | Mammalian | BSL-2 | ATCC | HEK2025-01 |
| CHO-K1 | Mammalian | BSL-1 | Thermo Fisher | CHO2025-02 |
| E. coli DH5α | Prokaryotic | BSL-1 | NCIM | EC2025-05 |
Revision History:
| Revision Date | Revision No. | Revision Details | Reason for Revision | Approved By |
|---|---|---|---|---|
| 04/05/2025 | 2.0 | Revised and reissued SOP under version 2.0 | Annual review and update |