IN THIS WEBINAR YOU WILL LEARN:
- How Microfluidic Modulation Spectroscopy (MMS) with pressure-assisted protein refolding and pressure perturbation can be used to study protein stability in biopharmaceutical formulation development.
- The benefits and sensitivity of MMS for observing secondary structure in complex formulations
- How high hydrostatic pressure positively affects protein conformation
- How the combination of Pressure assisted refolding, and MMS can be applied to different aspects of biotherapeutic characterization
Some of the questions you'll get the answer to in this webinar:
- Can you comment on why you chose human IgG as a model for testing the synergy between pressure perturbation and MMS analysis?
- How much optimization of parameters is needed before measuring each sample or spectrum and for each sample set?
High hydrostatic pressure in protein secondary structure studies
High hydrostatic pressure acts as a thermodynamic parameter causing perturbation of protein structure. This can lead to the unfolding of proteins orthogonal to the action of temperature or various chaotropic reagents. Pressure effects on protein structure are caused by hydration of solvent-excluded cavities that are eliminated during protein unfolding. Proteins overexpressed in bacterial systems tend to be misfolded and aggregated in a way that forms larger cavities, compared to native protein structures. Therefore, pressurization leads to preferential solubilization of protein from inclusion bodies, aiding in purification. Pressure unfolding can also preserve some stable secondary structure features (such as alpha-helical motifs) that may already exist in recombinant protein trapped in inclusion bodies, potentially simplifying the subsequent refolding steps.
Infrared spectroscopy, circular dichroism spectrophotometry and fluorescence spectroscopy are popular methods for monitoring changes in protein secondary structure. However, each technique has limitations in measurable sample composition. Microfluidic Modulation Spectroscopy (MMS) provides high-resolution structural information, critical to understanding the effects of protein misfolding and aggregation on biotherapeutic drug products.
Analyzing Protein Stability in Biopharmaceutical Formulation
Scientists today are challenged in gathering a complete understanding of protein structure using available techniques. They want to monitor structural changes that can alter the drug during development. These changes have been historically difficult to detect, as traditional analytical techniques are poor at detecting small differences in protein structure. MMS, however, can detect these changes with improved sensitivity and accuracy without the need for dilution or buffer exchange. In this GEN webinar, we will review pressure-assisted protein refolding and pressure perturbation coupled with MMS to study protein stability in biopharmaceutical formulations.