Advancing secondary structure characterization of monoclonal antibodies using Microfluidic Modulation Spectroscopy

Area of Science:

• Biophysical characterization
• Spectroscopic analysis
• Protein therapeutics

Background:

• Infrared (IR) spectroscopy is increasingly used for monitoring biotherapeutic critical quality attributes.
• Microfluidic Modulation Spectroscopy (MMS) is an automated IR technology for protein secondary structure analysis.
• MMS has demonstrated effectiveness in generating high-quality, reproducible data.

Purpose of the Study:

• To evaluate Microfluidic Modulation Spectroscopy (MMS) for analyzing the secondary structure of monoclonal antibodies (mAbs) at various concentrations.
• To optimize critical acquisition parameters for MMS to achieve high-quality data.
• To assess the reliability of MMS for low concentration biotherapeutic formulations.

Main Methods:

• Analysis of monoclonal antibodies (mAbs) across a concentration range of 0.5 to 50 mg/mL.
• Optimization of two key acquisition parameters: sample modulation frequency and detector dwell time.
• Utilized Microfluidic Modulation Spectroscopy (MMS), a novel automated IR technology.

Main Results:

• High-quality, reproducible secondary structure data was obtained for mAbs.
• Successful optimization of modulation frequency and detector dwell time settings.
• Demonstrated high sensitivity and reproducibility at low formulation concentrations (0.5-50 mg/mL).

Conclusions:

• Microfluidic Modulation Spectroscopy (MMS) is a reliable method for evaluating the secondary structure of biotherapeutic formulations.
• MMS is particularly effective for analyzing low concentration protein therapeutics and modalities.
• The optimized parameters enable robust secondary structure monitoring of monoclonal antibodies.