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Solvent‐free liquid avidin as a step toward cold chain elimination
Author(s) -
BuiLe Liem,
Brogan Alex P. S.,
Hallett Jason P.
Publication year - 2021
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.27587
Subject(s) - circular dichroism , avidin , chemistry , dynamic light scattering , denaturation (fissile materials) , thermal stability , protein secondary structure , analytical chemistry (journal) , crystallography , biophysics , materials science , chromatography , nanotechnology , organic chemistry , nuclear chemistry , biochemistry , biotin , biology , nanoparticle
The temperature sensitivity of vaccines and therapeutic proteins forces the distribution of life‐saving treatments to rely heavily on the temperature‐controlled (usually 2–8°C) supply and distribution network known as the cold chain. Here, using avidin as a model, we demonstrate how surface engineering could significantly increase the thermal stability of therapeutic proteins. A combination of spectroscopic (Fourier transform infrared, circular dichroism, and ultraviolet‐visible) and scattering techniques (dynamic light scattering, small‐angle, and wide‐angle X‐ray scattering) were deployed to probe the activity, structure, and stability of the model protein. Temperature‐dependent synchrotron radiation circular dichroism spectroscopy was used to demonstrate a significant increase in thermal stability, with a half denaturation temperature of 139.0°C and reversible unfolding with modified avidin returning to a 90% folded state when heated to temperatures below 100°C. Accelerated aging studies revealed that modified avidin retained its secondary structure after storage at 40°C for 56 days, equivalent to 160 days at 25°C. Furthermore, binding studies with multiple ligands revealed that the binding site remained functional after modification. As a result, this approach has potential as a storage technology for therapeutic proteins and the elimination of the cold chain, enabling the dissemination of life‐saving vaccines worldwide.