Standard Operating Procedure for Microbial Contamination Monitoring in Biosimilar Laboratories

Department	Biosimilars
SOP No.	SOP/BS/043/2025
Supersedes	SOP/BS/043/2022
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 routine monitoring and control of microbial contamination in biosimilar R&D environments, including clean areas, cell culture labs, and equipment, to ensure sterility, safety, and compliance with GMP guidelines.

2. Scope

This SOP applies to all areas handling live cells, media, reagents, and materials used in biosimilar development where microbial control is critical, including incubators, biosafety cabinets, and storage areas.

3. Responsibilities

• Microbiology Personnel: Perform sampling, record results, and escalate deviations.
• QA Officer: Review logs, initiate corrective actions, and trend results monthly.
• Lab Technicians: Maintain aseptic conditions and support follow-up actions.

4. Accountability

The Microbiology Lab Head is accountable for ensuring effective contamination monitoring and immediate response to microbial deviations in all R&D and production support zones.

5. Procedure

5.1 Environmental Monitoring Plan

1. Develop a monitoring map for:
   • Air (settle plates)
   • Surface (contact plates, swabs)
   • Personnel (glove/finger dabs)
2. Define frequencies:
   • Daily for clean rooms (Class 1000)
   • Weekly for non-critical zones
   • Monthly trending of all data

5.2 Sample Collection

1. Air Sampling: Use 90 mm settle plates (SDA for fungi, TSA for bacteria), expose for 4 hours.
2. Surface Swabbing: Use sterile swabs moistened with saline, cover 25 cm² area.
3. Personnel Sampling: Use contact plates on gloved fingertips before and after work.

5.3 Incubation and Identification

1. Incubate plates at:
   • 30–35°C for 48 hrs (bacteria)
   • 20–25°C for 5 days (fungi)
2. Record colony counts in the Microbial Monitoring Log (Annexure-1).
3. Identify isolates above alert level using Gram staining and biochemical methods.

5.4 Alert and Action Limits

1. Set based on area classification:
   • Air: ≤10 CFU/plate (Class B)
   • Surface: ≤5 CFU/plate
   • Personnel: ≤2 CFU/plate
2. Exceeding limits triggers investigation (Annexure-2).

5.5 Corrective and Preventive Actions (CAPA)

1. Initiate CAPA form and conduct:
   • Root cause analysis
   • Re-sanitization
   • Re-training of personnel
2. Verify effectiveness through follow-up sampling within 24–48 hours.

5.6 Trend Analysis

1. Monthly graphical analysis of microbial data by area and sample type.
2. Identify recurring hotspots and recommend control improvements.

6. Abbreviations

• CFU: Colony Forming Units
• SDA: Sabouraud Dextrose Agar
• TSA: Tryptic Soy Agar
• CAPA: Corrective and Preventive Action

7. Documents

1. Microbial Monitoring Log (Annexure-1)
2. Microbial Deviation Report (Annexure-2)
3. CAPA Form (Annexure-3)

8. References

• WHO TRS 961 Annex 6 – GMP for Biological Products
• EU GMP Annex 1 – Manufacture of Sterile Medicinal Products
• USP <797> and <1116> – Microbial Control Guidelines

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name
Designation
Department

11. Annexures

Annexure-1: Microbial Monitoring Log

Date	Location	Sample Type	Media	CFU Count	Result	Technician
02/05/2025	BSL-2 Lab	Surface	TSA	2	Pass	Sunita Reddy

Annexure-2: Microbial Deviation Report

Date	Area	Deviation	CFU Value	Initial Action	Investigated By
01/05/2025	Incubator #2	Surface > Limit	7	Disinfected	Ajay Verma

Annexure-3: CAPA Form

CAPA ID	Root Cause	Corrective Action	Preventive Action	Status
CAPA-043-01	Glove puncture	Re-trained user	Daily glove inspection	Closed

Revision History:

Revision Date	Revision No.	Revision Details	Reason for Revision	Approved By
04/05/2025	2.0	Updated alert/action limits and included trend analysis section	GMP Update