Standard Operating Procedure for Chromatography Column Cleaning Before Reuse in Biosimilar Manufacturing
| Department | Biosimilars |
|---|---|
| SOP No. | SOP/BS/135/2025 |
| Supersedes | SOP/BS/135/2022 |
| Page No. | Page 1 of 11 |
| Issue Date | 04/05/2025 |
| Effective Date | 06/05/2025 |
| Review Date | 04/05/2026 |
1. Purpose
To establish a validated procedure for cleaning chromatography columns used in biosimilar purification prior to reuse, in order to remove residual product, prevent cross-contamination, and maintain resin performance as per GMP and manufacturer’s guidelines.
2. Scope
This SOP applies to Protein A, ion exchange, hydrophobic interaction, and size exclusion chromatography columns used in the downstream processing (DSP) of biosimilar products in GMP manufacturing areas.
3. Responsibilities
- DSP Operators: Execute column cleaning procedures and log cleaning steps.
- Engineering: Maintain cleaning skid/systems and verify flowrates, temperature, and pressure parameters.
- QA: Review cleaning logs and verify cleaning validation or routine status before next column use.
4. Accountability
The Head of Manufacturing is accountable for ensuring chromatography column cleaning procedures are performed as per validated protocols and that logs are completed and approved before reuse.
5. Procedure
5.1 Pre-Cleaning Inspection
- Confirm cleaning is required post purification run.
- Verify the cleaning method (CIP/manual) as per the column type and resin specification.
- Check column ID, resin type, batch number, and previous use log (Annexure-1).
5.2 Column Cleaning Steps
- Attach column to cleaning skid or validated cleaning system.
- Flush with WFI or buffer until UV baseline stabilizes (typically 5–10 CV).
- Use appropriate cleaning agents depending on resin type:
- Protein A: 0.5 M NaOH, hold for 30 min contact time
- IEX: 1 M NaCl followed by 0.5 M NaOH
- SEC: 0.1 M NaOH or 20% ethanol (for microbial control)
- Flush column with WFI until pH and conductivity reach neutral range (pH 6.0–8.0).
- Record volumes, contact times, and final UV/conductivity readings in Annexure-2.
5.3 Sanitization and Storage
- For long-term storage, sanitize with 20% ethanol or 0.1 M NaOH as per resin compatibility.
- Seal column ends and affix storage label with:
- Last cleaning date
- Next due date
- Resin lifetime usage (number of cycles)
5.4 Cleaning Validation/Verification
- Initial 3 cleaning cycles post-purchase shall be validated using:
- TOC (Total Organic Carbon)
- Protein assay (BCA or Lowry)
- Microbial swabs
- Routine verification can be performed every 5 cycles and documented in Annexure-3.
5.5 Documentation
- Ensure all cleaning records are updated in the Chromatography Column Cleaning Log (Annexure-4).
- QA review and approval must be completed before next batch allocation.
6. Abbreviations
- CIP: Clean-In-Place
- CV: Column Volume
- DSP: Downstream Processing
- QA: Quality Assurance
7. Documents
- Column Usage Log – Annexure-1
- Column Cleaning Record – Annexure-2
- Cleaning Validation Report – Annexure-3
- Column Cleaning Master Log – Annexure-4
8. References
- Resin Manufacturer Cleaning Instructions
- ICH Q7 – GMP for Active Pharmaceutical Ingredients
- EU GMP Annex 15 – Qualification and Validation
9. SOP Version
Version: 2.0
10. Approval Section
| Prepared By | Checked By | Approved By | |
|---|---|---|---|
| Signature | |||
| Date | |||
| Name | |||
| Designation | |||
| Department |
11. Annexures
Annexure-1: Column Usage Log
| Column ID | Resin | Batch Used | Cycle No. | Last Cleaning Date |
|---|---|---|---|---|
| COL-001 | Protein A | BS-DSP-042 | 18 | 01/05/2025 |
Annexure-2: Column Cleaning Record
| Date | Cleaning Agent | Volume (L) | Contact Time (min) | Rinse pH | Conductivity |
|---|---|---|---|---|---|
| 04/05/2025 | 0.5 M NaOH | 10 | 30 | 7.2 | 5.1 µS/cm |
Annexure-3: Cleaning Validation Report
| Date | TOC | Protein | Microbial | Status |
|---|---|---|---|---|
| 04/05/2025 | Pass | Pass | Pass | Validated |
Annexure-4: Column Cleaning Master Log
| Column ID | Date | Cycle No. | Performed By | QA Reviewed |
|---|---|---|---|---|
| COL-001 | 04/05/2025 | 19 | Ajay Verma | Sunita Reddy |
Revision History:
| Revision Date | Revision No. | Revision Details | Reason for Revision | Approved By |
|---|---|---|---|---|
| 04/05/2025 | 2.0 | Added sanitization step and validation frequency | Process Enhancement |