Standard Operating Procedure for Column Packing and Unpacking in Biosimilar Manufacturing

Department	Biosimilars
SOP No.	SOP/BS/165/2025
Supersedes	SOP/BS/165/2022
Page No.	Page 1 of 10
Issue Date	04/05/2025
Effective Date	06/05/2025
Review Date	04/05/2026

1. Purpose

To describe the procedure for packing and unpacking chromatography columns used in biosimilar purification under GMP conditions, ensuring column efficiency, reproducibility, and resin integrity.

2. Scope

This SOP applies to all preparative chromatography columns (glass or stainless steel) used in downstream processing at lab or manufacturing scale for biosimilar monoclonal antibody or protein purification.

3. Responsibilities

• Production: Perform packing/unpacking as per validated protocols and record all observations.
• QA: Review packing logs and approve column integrity before use.
• Engineering: Ensure equipment readiness and flow control system calibration.

4. Accountability

The Head of Downstream Manufacturing is accountable for maintaining column performance and compliance with GMP during all packing and unpacking activities.

5. Procedure

5.1 Column Packing

1. Preparation:
   • Inspect and clean column housing, frits, adaptors, and seals.
   • Ensure the packing station and packing buffer are ready.
2. Slurry Preparation:
   • Mix resin with packing buffer (1:2 to 1:3 ratio).
   • Degas and allow settling for uniform distribution.
3. Packing Process:
   • Close bottom outlet, pour slurry into the column tube.
   • Attach the top adapter gently and allow air to escape.
   • Initiate flow at recommended flow rate (e.g., 300 cm/hr) and pack under gravity or pump pressure.
4. Compression and Stabilization:
   • Move adapter slowly to the settled bed and lock in place.
   • Flow equilibration buffer at operating flow rate and monitor pressure.
   • Record asymmetry and plate height to assess packing efficiency.

5.2 Column Testing

1. Run a test with 1% acetone or NaCl solution to verify asymmetry and theoretical plates.
2. Acceptance Criteria:
   • Asymmetry factor: 0.8–1.8
   • Number of theoretical plates: >2000 per meter

5.3 Column Unpacking

1. Disconnect column from skid and drain buffer from resin bed.
2. Remove top adapter and slowly release slurry into disposal vessel or clean container for reuse.
3. Rinse column tube and all parts with WFI and store as per cleaning SOP.
4. Log unpacking date and any damage to column or frits in Annexure-2.

5.4 Post-Packing Documentation

1. Record:
   • Resin lot number
   • Column ID and dimensions
   • Packing buffer details
   • Bed height, asymmetry, theoretical plates
2. Attach chromatogram and packing test results in Annexure-3.

6. Abbreviations

• GMP: Good Manufacturing Practices
• WFI: Water for Injection
• SEC: Size Exclusion Chromatography
• CV: Column Volume

7. Documents

1. Column Packing Record – Annexure-1
2. Column Unpacking Record – Annexure-2
3. Packing Evaluation Report – Annexure-3

8. References

• ICH Q8 – Pharmaceutical Development
• WHO TRS 999 – GMP Guidelines for Biologics
• OEM Manuals – Cytiva, Pall, and Millipore Chromatography Columns

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 Packing Record

Column ID	Resin Lot No.	Bed Height	Packing Buffer	Flow Rate	Date	Operator
COL-102	SP-9802	20 cm	20 mM Phosphate	300 cm/hr	04/05/2025	Ajay Verma

Annexure-2: Column Unpacking Record

Column ID	Date	Resin Recovered?	Condition	Remarks	Operator
COL-102	04/05/2025	Yes	Good	No cracks	Sunita Reddy

Annexure-3: Packing Evaluation Report

Column ID	Asymmetry	Theoretical Plates	Pass/Fail	Analyst	Date
COL-102	1.1	2850/m	Pass	Vinay Pawar	04/05/2025

Revision History:

Revision Date	Revision No.	Details	Reason	Approved By
04/05/2025	2.0	Added detailed resin testing and asymmetry range	Process harmonization and QA audit recommendation