Real-time detection of mAb aggregates in an integrated downstream process

Area of Science:

• Biopharmaceutical Manufacturing
• Process Analytical Technology (PAT)
• Microfluidics

Background:

• Continuous processing in biopharma requires real-time monitoring tools.
• Protein aggregation is a critical quality attribute that needs accurate measurement.
• Miniaturized sensors offer faster analysis and quicker decision-making.

Purpose of the Study:

• To integrate and assess a miniaturized fluorescent dye-based sensor in a continuous downstream bioprocess.
• To evaluate the sensor's capability for real-time detection of monoclonal antibody (mAb) aggregation.
• To demonstrate improved process understanding and control through at-line PAT implementation.

Main Methods:

• A microfluidic micromixer utilizing fluorescent dyes (Bis-ANS, CCVJ) was employed.
• The sensor was integrated into a lab-scale ÄKTA™ continuous downstream purification system.
• Viral inactivation and polishing steps were reproduced, with samples analyzed by the sensor post-processing.

Main Results:

• The sensor successfully detected protein aggregation levels as low as 2.5%.
• At-line measurements were completed in under 10 minutes, providing rapid feedback.
• Integration into the continuous process allowed for real-time monitoring of aggregation during purification.

Conclusions:

• The miniaturized PAT tool enables fast, at-line aggregation detection in continuous biopharmaceutical manufacturing.
• This technology enhances process understanding and facilitates better control over product quality.
• The developed system represents a significant step towards implementing robust PAT for continuous bioprocessing.