Standard Operating Procedure for Aseptic Handling During Inoculation in Biosimilar Manufacturing

Department	Biosimilars
SOP No.	SOP/BS/108/2025
Supersedes	SOP/BS/108/2022
Page No.	Page 1 of 12
Issue Date	04/05/2025
Effective Date	06/05/2025
Review Date	04/05/2026

1. Purpose

To outline the aseptic techniques and procedural steps required during inoculation of seed cultures into production bioreactors in biosimilar upstream processes, thereby minimizing the risk of contamination.

2. Scope

This SOP applies to all inoculation activities performed in cleanroom Grade B and C classified areas where seed cultures are transferred to bioreactors within the biosimilars manufacturing facility.

3. Responsibilities

• Production Operator: Perform inoculation aseptically and maintain documentation.
• Supervisor: Verify readiness of systems and approve inoculation process.
• QA: Oversee compliance with aseptic practices and review batch records.

4. Accountability

The Head of Production is accountable for ensuring that inoculation procedures are carried out aseptically and in compliance with GMP standards.

5. Procedure

5.1 Preparation and Environment Setup

1. Ensure the inoculation area is cleaned and disinfected using 70% IPA or validated biocide.
2. Verify HVAC parameters (temperature, humidity, pressure differentials) meet specifications.
3. Perform area clearance and fill Annexure-1: Pre-Inoculation Checklist.

5.2 Personnel Preparation

1. Only trained personnel qualified in aseptic handling may participate.
2. Perform full Grade B gowning procedure:
   • Sterile coveralls, masks, goggles, gloves, and footwear
3. Disinfect gloves periodically during handling.

5.3 Equipment and Material Preparation

1. Sanitize all transfer tubing, connectors, and clamps using 70% IPA.
2. Ensure seed bioreactor and production bioreactor are within validated temperature and DO ranges.
3. Verify seed culture viability (≥90%) and VCD before inoculation.

5.4 Aseptic Inoculation Procedure

1. Connect sterile tubing from seed culture vessel to the inoculation port of the bioreactor using aseptic connectors (e.g., AseptiQuik).
2. Initiate transfer using peristaltic pump under laminar flow or closed transfer system.
3. Monitor for any air bubble formation or leakage.
4. Upon completion, clamp all lines and disconnect using sterile scissors or welders.

5.5 Post-Inoculation Activities

1. Close all access ports and start bioreactor agitation and sparging per process parameters.
2. Document inoculation details in Annexure-2: Inoculation Record.
3. Send representative culture sample to QC for sterility and viability testing.

6. Abbreviations

• IPA: Isopropyl Alcohol
• VCD: Viable Cell Density
• QC: Quality Control
• GMP: Good Manufacturing Practice

7. Documents

1. Pre-Inoculation Checklist – Annexure-1
2. Inoculation Record – Annexure-2

8. References

• WHO TRS 961 – Annex 6: GMP for Biological Products
• EU GMP Annex 1 – Manufacture of Sterile Medicinal Products

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name
Designation
Department

11. Annexures

Annexure-1: Pre-Inoculation Checklist

Parameter	Status	Remarks	Checked By
Area Sanitized	Yes	–	Ajay Verma
HVAC Parameters	Within range	23°C, 50% RH	Sunita Reddy

Annexure-2: Inoculation Record

Date	Seed VCD	Viability (%)	Volume Transferred (L)	Bioreactor ID	Performed By
04/05/2025	1.8 x 10⁶	94	4.0	BR-1500A	Sunita Reddy

Revision History:

Revision Date	Revision No.	Revision Details	Reason for Revision	Approved By
04/05/2025	2.0	Added aseptic connector steps and updated gowning standards	Regulatory update