Standard Operating Procedure for Media Preparation and Sterilization in Biosimilar Upstream Processing

Department	Biosimilars
SOP No.	SOP/BS/062/2025
Supersedes	SOP/BS/062/2022
Page No.	Page 1 of 12
Issue Date	04/05/2025
Effective Date	06/05/2025
Review Date	04/05/2026

1. Purpose

To define the procedure for accurate preparation, sterilization, and storage of culture media used in upstream biosimilar fermentation processes to ensure product quality, batch consistency, and GMP compliance.

2. Scope

This SOP applies to the formulation and sterilization of both basal and feed media used in shake flask cultures and bioreactors during biosimilar production, including chemically defined and serum-free media.

3. Responsibilities

• Upstream Technicians: Execute media preparation and sterilization steps as per protocol.
• QA: Review preparation logs and conduct environmental and sterility checks.
• Warehouse Team: Supply media components and verify expiry and label integrity.

4. Accountability

The Head of Upstream Process is accountable for ensuring media is prepared and sterilized under controlled conditions and that all steps are documented and traceable.

5. Procedure

5.1 Raw Material Verification

1. Ensure all raw materials (powdered media, water, buffers) are available as per Material Requisition Form.
2. Verify CoA, expiry date, and storage conditions of each item before use (Annexure-1).

5.2 Media Preparation

1. Clean and sanitize media preparation area before starting.
2. Weigh media components using calibrated balance; record batch-wise details in Media Preparation Log (Annexure-2).
3. Dissolve media powder in Water for Injection (WFI) using magnetic stirrers or overhead stirrers as applicable.
4. Adjust pH using 1N NaOH or 1N HCl to required range (e.g., 6.8–7.2).
5. Top up final volume and ensure uniform mixing.

5.3 Filtration and Sterilization

1. Filter media using 0.22 µm sterilizing-grade filters into pre-sterilized containers under laminar flow hood.
2. Label media containers with batch number, name, pH, date, and storage condition (Annexure-3).
3. If heat sterilization is required, autoclave at 121°C for 30 minutes unless otherwise specified.

5.4 Storage and Usage

1. Store sterilized media at 2–8°C or room temperature as specified by the manufacturer or protocol.
2. Media must be used within validated hold time period (refer to SOP/BS/099/2025).
3. Inspect for clarity, pH drift, or contamination before use.

5.5 Cleaning and Documentation

1. Clean all equipment post-use with validated cleaning solution (e.g., 1% NaOH followed by WFI rinse).
2. Record preparation, sterilization, and cleaning activities in Media Use and Cleaning Log (Annexure-4).

6. Abbreviations

• WFI: Water for Injection
• pH: Potential of Hydrogen
• GMP: Good Manufacturing Practice
• CoA: Certificate of Analysis

7. Documents

1. Raw Material Verification Checklist – Annexure-1
2. Media Preparation Log – Annexure-2
3. Media Label Record – Annexure-3
4. Media Use and Cleaning Log – Annexure-4

8. References

• ICH Q7 – GMP for Active Pharmaceutical Ingredients
• WHO TRS 1010 – Biopharmaceutical Production
• 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: Raw Material Verification Checklist

Material	Batch No.	Expiry	Storage Verified	Checked By
CD CHO Powder	MP-CH/0425	06/2026	Yes	Sunita Reddy

Annexure-2: Media Preparation Log

Date	Media Type	Volume Prepared (L)	pH	Operator
03/05/2025	Feed A	10	7.0	Ajay Verma

Annexure-3: Media Label Record

Container ID	Media Type	Preparation Date	Expiry Date	Storage
FLK001	Basal Media	03/05/2025	10/05/2025	2–8°C

Annexure-4: Media Use and Cleaning Log

Date	Media Used	Equipment Cleaned	Cleaning Agent	Verified By
04/05/2025	Feed A	Tank-3	1% NaOH	QA Officer

Revision History:

Revision Date	Revision No.	Revision Details	Reason for Revision	Approved By
04/05/2025	2.0	Included filtration criteria and pH adjustment parameters	Periodic SOP Review