Standard Operating Procedure for Nutrient Feed Preparation and Addition in Bioreactor Operations for Biosimilars

Department	Biosimilars
SOP No.	SOP/BS/076/2025
Supersedes	SOP/BS/076/2022
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 standardized procedure for the preparation, filtration, labeling, storage, and controlled aseptic addition of nutrient feeds into bioreactors during biosimilar upstream processing.

2. Scope

This SOP applies to all types of nutrient feeds (e.g., glucose, amino acids, lipids, trace elements) prepared for use in fed-batch and perfusion bioreactor processes in the biosimilar production facility.

3. Responsibilities

• Process Operators: Prepare, filter, and aseptically transfer nutrient feeds as per instructions.
• Formulation Chemist: Verify feed formulations and ensure raw material identity and quantity.
• QA Personnel: Approve feed preparation records and cross-check batch details.

4. Accountability

The Head of Upstream Manufacturing is accountable for ensuring nutrient feeds are prepared and added correctly, without compromising sterility or cell culture performance.

5. Procedure

5.1 Raw Material Verification

1. Verify all raw materials against the Bill of Materials (BoM) and ensure valid CoA is available.
2. Ensure containers are labeled with item code, batch number, and expiry date.

5.2 Feed Preparation

1. Transfer materials into the formulation area under laminar airflow.
2. Use calibrated balances to weigh materials with ±0.1 g accuracy (Annexure-1).
3. Dissolve powders in WFI or process-grade water with magnetic stirring or overhead mixer.
4. Adjust pH (if required) and document final pH before filtration.

5.3 Sterile Filtration

1. Filter solution through a 0.22 µm PES or PVDF sterile-grade filter into a sterile, labeled container.
2. Conduct filter integrity test pre- and post-use (Annexure-2).

5.4 Labeling and Storage

1. Label containers with Feed Type, Batch No., Preparation Date, Expiry Date, Volume, and Storage Conditions (Annexure-3).
2. Store feed solutions between 2–8°C for up to 7 days or as validated.

5.5 Feed Addition to Bioreactor

1. Bring feed solution to ambient temperature before use.
2. Disinfect container surface using 70% IPA and connect aseptically to bioreactor via sterile tubing or port.
3. Use peristaltic pump or gravity flow to add feed; avoid contamination or backflow.
4. Record feed volume, addition time, and operator initials (Annexure-4).

6. Abbreviations

• WFI: Water for Injection
• IPA: Isopropyl Alcohol
• PES: Polyethersulfone
• PVDF: Polyvinylidene Difluoride

7. Documents

1. Weighing and Formulation Sheet – Annexure-1
2. Filter Integrity Log – Annexure-2
3. Feed Label Template – Annexure-3
4. Feed Addition Record – Annexure-4

8. References

• ICH Q7 – GMP for Active Pharmaceutical Ingredients
• WHO TRS 1025 – Biomanufacturing Guidelines
• USP <797> – Sterile Preparations

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name
Designation
Department

11. Annexures

Annexure-1: Weighing and Formulation Sheet

Material	Required Qty (g)	Actual Qty (g)	Weighed By	Date
Glucose Monohydrate	100	100.01	Rajesh Kumar	04/05/2025

Annexure-2: Filter Integrity Log

Filter ID	Pre-Test Result	Post-Test Result	Passed?	Checked By
FLT-076-01	Pass	Pass	Yes	Sunita Reddy

Annexure-3: Feed Label Template

Feed Type	Batch No.	Volume	Prep Date	Expiry	Storage
Feed B	NF-BIO-076-01	3L	04/05/2025	11/05/2025	2–8°C

Annexure-4: Feed Addition Record

Date	Time	Volume Added (mL)	Operator Initials	Remarks
04/05/2025	14:00	150	RK	Initial bolus feed

Revision History:

Revision Date	Revision No.	Revision Details	Reason for Revision	Approved By
04/05/2025	2.0	Added filtration and labeling procedures	GMP alignment