Standard Operating Procedure for Use of Chemically Defined Media in Biosimilar Manufacturing

Department	Biosimilars
SOP No.	SOP/BS/063/2025
Supersedes	SOP/BS/063/2022
Page No.	Page 1 of 11
Issue Date	04/05/2025
Effective Date	06/05/2025
Review Date	04/05/2026

1. Purpose

To define the standardized procedure for selection, preparation, and use of chemically defined media (CDM) in upstream biosimilar manufacturing to ensure reproducibility, animal component-free operations, and alignment with regulatory expectations.

2. Scope

This SOP applies to all cell culture activities utilizing chemically defined media in biosimilar upstream processes including shake flasks, seed cultures, and bioreactors involving CHO, HEK293, and other mammalian cell lines.

3. Responsibilities

• Process Technologist: Responsible for selecting and validating CDM for specific cell lines.
• Upstream Operator: Prepares and handles CDM under aseptic conditions as per SOP.
• QA: Reviews documentation and verifies traceability and batch records.

4. Accountability

The Head of Upstream Process is accountable for the implementation of CDM and ensuring batch-wise consistency and GMP-compliance in upstream cell culture operations.

5. Procedure

5.1 Media Selection and Validation

1. Select CDM based on cell line compatibility and product titer objectives.
2. Perform bench-scale studies comparing growth and productivity metrics in at least two media brands.
3. Document comparative results in Media Evaluation Sheet (Annexure-1).

5.2 Preparation of CDM

1. Confirm lot release and CoA of selected media powder or liquid formulation.
2. Weigh and dissolve media in Water for Injection (WFI) using magnetic stirrers under cleanroom conditions.
3. Adjust pH (6.8–7.2) using sterile NaOH or HCl if required.
4. Filter through 0.22 µm sterile membrane filter into pre-labeled sterile vessels (Annexure-2).

5.3 Application in Cell Culture

1. Thaw cryovial as per SOP/BS/027/2025 and initiate culture in T-flask with CDM.
2. Monitor daily growth using VCD and viability via hemocytometer or cell counter.
3. Scale up seed cultures in shake flasks maintaining optimal aeration and temperature (37°C, 5% CO₂).

5.4 Monitoring and Hold Time

1. Check pH, osmolality, and sterility of CDM before each use.
2. Store CDM at 2–8°C and use within validated hold time as per SOP/BS/099/2025.
3. Dispose unused or expired media as per biosafety guidelines (SOP/BS/048/2025).

5.5 Documentation

1. Record all preparation and usage in Media Usage Log (Annexure-3).
2. Attach CoA, lot number, and QA release documents with batch record.

6. Abbreviations

• CDM: Chemically Defined Media
• WFI: Water for Injection
• VCD: Viable Cell Density
• GMP: Good Manufacturing Practice

7. Documents

1. Media Evaluation Sheet – Annexure-1
2. CDM Preparation and Label Record – Annexure-2
3. Media Usage Log – Annexure-3

8. References

• ICH Q5D – Derivation and Characterization of Cell Substrates
• WHO TRS 999 – Biotechnological Manufacturing Practices
• SOP/BS/099/2025 – Media Hold Time 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: Media Evaluation Sheet

Media Name	Growth Rate	Max VCD	Product Titer	Preferred?
CD CHO A	0.4/day	3.2 × 10⁶	120 mg/L	Yes
CD CHO B	0.3/day	2.8 × 10⁶	95 mg/L	No

Annexure-2: CDM Preparation and Label Record

Date	Media Type	Lot No.	pH	Filter ID	Operator
03/05/2025	CD CHO A	CC-A-0425	7.1	FT-022	Rajesh Kumar

Annexure-3: Media Usage Log

Date	Batch ID	Media Used	Volume (L)	Remarks
04/05/2025	SD-CHO-105	CD CHO A	5	Used for seed expansion

Revision History:

Revision Date	Revision No.	Revision Details	Reason for Revision	Approved By
04/05/2025	2.0	Inclusion of osmolality checks and evaluation template	Process Standardization