Standard Operating Procedure for Cleaning Validation of Single-Use Systems in Biosimilar Manufacturing

Department	Biosimilars
SOP No.	SOP/BS/138/2025
Supersedes	SOP/BS/138/2022
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 validation strategy and procedure for ensuring cleaning effectiveness of single-use systems (SUS) in biosimilar manufacturing operations and to confirm the absence of cross-contamination risks, extractables, and leachables post-rinsing and usage.

2. Scope

This SOP applies to disposable bioreactor bags, tubing sets, mixers, filters, and bags used in upstream and downstream processing where reuse is practiced under restricted and validated conditions (e.g., tubing manifolds or connector assemblies).

3. Responsibilities

• Production: Execute cleaning or rinsing procedures on designated SUS components.
• QC (Chemical & Microbiology): Perform analytical and microbial testing for residues and sterility assurance.
• QA: Approve cleaning validation protocols and reports and verify compliance.

4. Accountability

The QA Head is accountable for ensuring validated cleaning protocols for all SUS used in GMP operations, aligned with risk-based control strategies and regulatory expectations.

5. Procedure

5.1 Identification of Reusable SUS

1. Define SUS components eligible for reuse based on:
   • Vendor recommendations
   • Process contact risk
   • Regulatory acceptance (e.g., < 3 reuse cycles)
2. Document these in Annexure-1: SUS Reuse Assessment Form.

5.2 Cleaning/Rinsing Procedure

1. Post-use, rinse the SUS component with WFI or buffer (3x volume flush) using closed-loop system.
2. Perform visual inspection for particulates, discoloration, or fouling.
3. Capture rinse samples in pre-cleaned containers for TOC, conductivity, and bioburden testing.

5.3 Validation Strategy

1. Design cleaning validation protocol considering:
   • Worst-case product load
   • Sampling technique (direct contact or rinse)
   • Analytical detection limits
2. Execute 3 consecutive successful cleanings for each SUS category.
3. Maintain data in Annexure-2: Cleaning Validation Log.

5.4 Acceptance Criteria

1. TOC: ≤ 10 ppm
2. Conductivity: Within 10% of WFI baseline
3. Bioburden: < 10 CFU/100 mL
4. No visible residue or blockage

5.5 Documentation and Approval

1. All validation activities must be documented in the Cleaning Validation Summary Report.
2. QA must sign off validation and issue a status label/tag on SUS (Valid for Reuse).

6. Abbreviations

• SUS: Single-Use System
• TOC: Total Organic Carbon
• CFU: Colony Forming Units
• QA: Quality Assurance
• QC: Quality Control

7. Documents

1. SUS Reuse Assessment Form – Annexure-1
2. Cleaning Validation Log – Annexure-2

8. References

• FDA Guidance for Industry: Single-Use Technology
• EU GMP Annex 15 – Qualification and Validation
• USP <665>, <1665> – Extractables and Leachables

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name
Designation
Department

11. Annexures

Annexure-1: SUS Reuse Assessment Form

SUS ID	Component Type	Risk Level	Reuse Limit	Approval Status
SUS-001	Transfer Manifold	Low	3	Approved

Annexure-2: Cleaning Validation Log

Date	SUS ID	TOC (ppm)	Conductivity (µS/cm)	Bioburden (CFU)	Result
03/05/2025	SUS-001	4.2	1.5	0	Pass

Revision History:

Revision Date	Revision No.	Revision Details	Reason for Revision	Approved By
04/05/2025	2.0	Included validation sampling and E&L references	Regulatory Update