Standard Operating Procedure for Inline Analytics (Raman, NIR) in Bioreactor Operations for Biosimilars

Department	Biosimilars
SOP No.	SOP/BS/096/2025
Supersedes	SOP/BS/096/2022
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 procedure for using inline Raman and Near Infrared (NIR) spectroscopy systems in bioreactor operations for real-time monitoring of critical process parameters (CPPs) in biosimilar manufacturing.

2. Scope

This SOP applies to inline Raman and NIR analyzers used in pilot-scale and production-scale bioreactors for monitoring glucose, lactate, amino acids, and other metabolic markers.

3. Responsibilities

• Process Analytical Technology (PAT) Team: Configure instruments, develop models, and validate performance.
• Manufacturing Operator: Ensure probe integrity and monitor real-time analytics during batch processing.
• QA Department: Review calibration and performance qualification (PQ) reports.

4. Accountability

The Head of Manufacturing is accountable for the proper implementation and maintenance of inline Raman and NIR systems in compliance with GMP and QbD principles.

5. Procedure

5.1 Installation and Pre-Use Checks

1. Verify inline probe installation in designated ports on bioreactor lid or sampling loops.
2. Ensure cleaning and sterilization of probe are completed prior to use (refer CIP/SIP procedures).
3. Confirm firmware, spectral libraries, and calibration models are current and approved.

5.2 Instrument Startup

1. Power on Raman/NIR analyzer using dedicated PAT console.
2. Initialize spectral acquisition software (e.g., SynTQ, Empower PAT, etc.).
3. Verify baseline spectra with sterile WFI or media blank.

5.3 Model Calibration and Verification

1. Use standard solutions or reference materials to verify predictive models (e.g., glucose 5 g/L).
2. Acceptable prediction error (RMSEP) must be ≤10% of target concentration.
3. Document spectral match and model performance in Annexure-1.

5.4 Real-Time Monitoring During Batch

1. Acquire spectra at 10-minute intervals or as defined in process protocol.
2. Monitor CPPs such as:
   • Glucose, Lactate
   • pH, Ammonia
   • Viable cell density (if enabled)
3. Set alarms in software for critical threshold deviations.

5.5 Post-Use Data Handling

1. Export spectral data and process trends to secure server.
2. Generate report including:
   • Parameter trends
   • Sensor stability
   • Deviation records

5.6 Maintenance and Requalification

1. Clean probes post-use with validated cleaning solution (e.g., 1M NaOH followed by WFI rinse).
2. Perform annual calibration and spectral model revalidation.
3. Maintain all logs and attach PQ summary to Annexure-2.

6. Abbreviations

• PAT: Process Analytical Technology
• CPP: Critical Process Parameter
• RMSEP: Root Mean Square Error of Prediction
• WFI: Water for Injection
• QbD: Quality by Design

7. Documents

1. Inline Analytics Model Verification Log – Annexure-1
2. PAT Instrument Qualification Summary – Annexure-2

8. References

• ICH Q8(R2) – Pharmaceutical Development
• FDA Guidance on Process Analytical Technology (PAT) 2004

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name
Designation
Department

11. Annexures

Annexure-1: Inline Analytics Model Verification Log

Date	Probe ID	Analyte	Expected Value	Predicted Value	Deviation (%)	Tested By
04/05/2025	Raman-01	Glucose	5.0 g/L	5.2 g/L	+4%	Ajay Verma

Annexure-2: PAT Instrument Qualification Summary

Instrument	Serial No.	PQ Date	Next Due	Status	Verified By
NIR Analyzer	NIR-BIO-07	01/04/2025	01/04/2026	Qualified	Rajesh Kumar

Revision History:

Revision Date	Revision No.	Revision Details	Reason for Revision	Approved By
04/05/2025	2.0	Included RMSEP validation and alarm configuration step	Enhanced PAT compliance