Detection of Pressure-Induced Protein Aggregation Using Microfluidic Modulation Spectroscopy (MMS)

APPLICATION NOTE

The ability to detect protein aggregation is important at all stages of drug development. Early detection of aggregation is most desirable to inform development decisions since aggregation can negatively affect the functionality of a protein.

This Application Note Includes:

• Highlights the significant changes present in the spectral regions of 1624 and 1640cm-1 in the Second Derivative plot that can be seen only subtly in the Absolute Absorbance spectral plot.
• Weighted Spectral Difference (WSD) as a metric used to monitor change in sample secondary structure for this aggregation study
• Strong linear correlation between increasing amounts of percent aggregation and increasing amounts of measured antiparallel beta-sheet content

Pressure-Induced Protein Aggregation Detection
Protein aggregation is a recognized signal of instability and can lead to the loss of protein function. It is therefore crucial to detect protein aggregation early in the drug development process to inform further drug development decisions. Pressure, a stressor used for generating aggregates by impacting noncovalent interactions without the need to change temperature or solvents1, was employed to create aggregated protein for this spiked study.

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