Mid-infrared spectroscopy-based antibody aggregate quantification in cell culture fluids.

Area of Science:

• Biotechnology
• Analytical Chemistry
• Process Analytical Technology (PAT)

Background:

• Therapeutic antibody purification is prone to antibody aggregation.
• Current aggregate monitoring methods (chromatography, SDS-PAGE, light scattering) can be time-consuming or complex.
• Accurate aggregate quantification is crucial for ensuring therapeutic efficacy and safety.

Purpose of the Study:

• To investigate the feasibility of mid-infrared spectroscopy (MIR) for quantifying soluble antibody aggregates.
• To develop and validate multivariate models for aggregate quantification in purification streams and cell culture supernatants.
• To establish a generalizable MIR model for real-time aggregate monitoring.

Main Methods:

• Application of multivariate modeling techniques to mid-infrared spectral data.
• Analysis of antibody aggregation in chromatography elution streams and clarified CHO cell culture supernatants.
• Validation of the developed MIR model using real-process samples and process mimics with two different antibodies.

Main Results:

• A generalizable multivariate model was established for MIR-based aggregate quantification.
• The model demonstrated good prediction ability for aggregate content down to 1%.
• MIR provides a fast, cost-effective, and straightforward method for aggregate monitoring.

Conclusions:

• Mid-infrared spectroscopy is a suitable technique for quantifying soluble antibody aggregates in therapeutic antibody purification.
• MIR offers potential for multi-parameter process monitoring, including host cell proteins and target proteins, from a single measurement.
• This technology can enhance process understanding and control in biopharmaceutical manufacturing.