Infra-Red (IR) analysis has been long
accepted as a powerful tool in protein characterization, particularly in the
Amide I band (~1600 - 1700 cm-1), which gives detailed secondary-structural
information that can be critical in determining protein structure-activity
relationships, stability, batch-to-batch comparisons and in formulation studies
as a few examples. Technologies traditionally used for secondary structure analysis,
such as benchtop Fourier Transform IR (FTIR) or Circular Dichroism (CD), suffer
from a number of issues that have prevented their routine use in this area,
preventing this application from reaching its full potential. These include
concentration and buffer restrictions, incompatibility with a range of
excipients, a lack of automation, low spectral reproducibility and for FTIR,
water subtraction problems.
Microfluidic Modulation Spectroscopy (MMS) is a new key technology that was
brought to market in 2019 by RedShift Bioanalytics. It focuses on the IR Amide
I region to produce exceptionally high data quality and reproducibility that
aim to solve the aforementioned issues encountered with traditional
technologies. It is fully automated, running samples from 24- and 96-well
plates, compatible with a very broad concentration range (0.1 to >200
mg/ml), and is also compatible with a wide range of complex buffer systems and
excipients, including those that absorb in the amide I region, surfactants and
organic solvents. The platform includes a powerful software package that
facilitates data analysis, and can be included in the automation procedure.
This presentation highlights the technical benefits of MMS and its application
in the protein structural workflow, giving relevant application examples