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A Rapid and Quantitative Fluorimetric Method for Protein-Targeting Small Molecule Drug Screening
Published on: October 16, 2015
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Determining suitable surfactant concentration ranges to avoid protein unfolding in pharmaceutical formulations using
Laura J Waters1, Joseph Whiteley1, William Small2
1School of Applied Sciences, University of Huddersfield, Queensgate, Huddersfield, HD1 3DH, UK.
Heliyon
|November 13, 2023
Summary
This study introduces a UV spectroscopic method to detect protein unfolding in biologics. It found that protein unfolding begins at surfactant concentrations significantly below the critical micellar concentration, aiding formulation development.
Area of Science:
- Biopharmaceutical formulation
- Protein chemistry
- Analytical chemistry
Background:
- Protein stability is crucial for biologic drug efficacy.
- Maintaining the native 3D conformation is essential for therapeutic activity.
- Protein unfolding can lead to loss of function.
Purpose of the Study:
- To propose a modified UV spectroscopic method for detecting protein unfolding.
- To determine concentration limits for avoiding protein unfolding with polysorbate 20 and 80.
- To establish a screening tool for optimizing surfactant concentrations in protein formulations.
Main Methods:
- Utilized a modified UV spectroscopic technique.
- Correlated surfactant concentration with the percentage of unfolded protein for model proteins and surfactants (polysorbate 20 and 80).
- Identified distinct concentration regions related to protein unfolding.
Main Results:
- Two regions were observed: one with no unfolding, and a second with increasing unfolding.
- Protein unfolding commenced at concentrations approximately ten times lower than the critical micellar concentration.
- This pattern held true regardless of the specific protein or polysorbate used.
Conclusions:
- The adapted UV spectroscopic method can serve as a simple screening tool for formulation development.
- It helps identify the 'onset of unfolding' concentration for protein-surfactant systems.
- Optimizing surfactant concentration is key to maintaining biologic benefits while preventing inadvertent unfolding.
