A Fully Integrated Online Platform For Real Time Monitoring Of Multiple Product Quality Attributes In...

Area of Science:

• Biopharmaceutical Development
• Process Analytical Technology (PAT)
• Analytical Chemistry

Background:

• The FDA mandates Quality by Design (QbD) for biopharmaceutical development, requiring robust monitoring of critical quality attributes (CQAs).
• Previous offline Multi-Attribute Methods (MAM) demonstrated high sensitivity for monitoring over 20 CQAs in upstream bioprocesses.
• Laborious manual sampling and sample preparation hinder the routine application of offline methods.

Purpose of the Study:

• To develop and validate a fully integrated, automated online platform for real-time bioprocess monitoring using MAM.
• To overcome the limitations of manual sampling and sample preparation associated with offline analytical methods.
• To enable routine application of MAM for enhanced bioprocess control and quality assurance.

Main Methods:

• Integration of Modular Automated Sampling Technology (MAST) for aseptic sampling.
• Utilization of multi-function Sequential Injection Analysis (SIA) for automated sample preparation.
• Implementation of Ultra-High-Performance Liquid Chromatography (UHPLC) coupled with High-Resolution Mass Spectrometry (HRMS) for analysis.
• Demonstration of continuous, automated daily monitoring over a 17-day cell culture process for a model monoclonal antibody (mAb).

Main Results:

• Successful demonstration of an end-to-end automated online MAM system for real-time bioprocess monitoring.
• Achieved comparable specificity and sensitivity to previously reported offline MAM analyses.
• Continuous automated monitoring of a 17-day cell culture process for a model mAb was successfully performed.
• The system offers potential to replace multiple conventional, lower-resolution offline methods.

Conclusions:

• The developed automated online MAM platform represents a significant advancement in bioprocess monitoring technology.
• This system facilitates routine application of MAM, enhancing real-time control and quality assurance in biopharmaceutical manufacturing.
• The online HPLC/HRMS platform is adaptable for downstream purification monitoring with minimal software adjustments.