Microfluidic Modulation Spectroscopy (MMS) Fills an Analytical Gap with a Lower LOQ for Measuring Protein Misfolds and Structural Similarity

Improving spectroscopic methods for measuring low level impurities in secondary structure is necessary to maintain confidence in a protein’s integrity during all phases of drug development

Application Note Introduction

Protein misfolds in secondary structure can occur during all phases of drug development. A protein misfold represents a structural impurity and at any level can result in changed efficacy and increase the potential for immunogenicity. Improving spectroscopic methods for measuring low level impurities in secondary structure is necessary to maintain confidence in a protein’s integrity during all phases of drug development. Common structural characterization methods such as FTIR and CD have known limitations in reproducibility and sensitivity which adversely increase the lowest level of quantitation (LOQ) achievable when measuring structural impurities and similarity. The MMS system developed by RedShiftBio is a new protein characterization method which generates reproducible high resolution measurements. 

Techniques Showcased in This Application Note Include:

  • In this App, a model protein system consisting of different secondary structures in the amide I region was prepared to measure and compare structural similarity using three characterization methods:  MMS, FTIR and circular dichroism (CD)
  • Generates reproducible replicate spectra at both the raw differential spectral level and second derivative level, with and without area normalization
  • Detects and measures protein misfolds with a lower LOQ 
  • Automatically generates a stable signal and virtually drift-free subtracted spectra