Matthew McGann and Alexander Lazarev
Using both pressure cycling technology and microfluidic modulation spectroscopy reveals the fate of misfolded proteins, says Pressure BioSciences and RedShiftBio
If therapeutic proteins are to be safe and efficacious, their misfolding and aggregation behaviors must be well understood. These behaviors are usually clarified through detailed structural characterization, particularly of the secondary structure of the protein, using established technologies such as conventional Fourier transform infrared (FTIR) spectroscopy and far-ultraviolet circular dichroism (far-UV CD). However, these technologies may clash with workflow requirements.
An alternative technology is microfluidic modulation spectroscopy (MMS). It is a next-generation technique that optimally shapes IR spectroscopy for biopharmaceutical development. MMS, then, can measure previously undetectable changes in protein structural attributes, changes that are critical to drug efficacy and quality.
Proteins typically misfold or aggregate in response to thermal stress or in the presence of chaotropic agents, molecules that reduce structural order by disrupting noncovalent bonds. The resulting structural change tends to be irreversible, which is suboptimal when it comes to studying the kinetics and thermodynamics of aggregation; reversible change enables more detailed investigation. Reversible misfolding and aggregate dissociation can be promoted by the application of high hydrostatic pressure, which makes pressure cycling technology (PCT) a valuable, orthogonal tool for aggregation studies.