Protecting activity of desiccated enzymes

Protein‐based biological drugs and many industrial enzymes are unstable, making them prohibitively expensive. Some can be stabilized by formulation with excipients, but most still require low temperature storage. In search of new, more robust excipients, we turned to the tardigrade, a microscopic an...

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Bibliographic Details
Published in:Protein science 2019-05, Vol.28 (5), p.941-951
Main Authors: Piszkiewicz, Samantha, Gunn, Kathryn H., Warmuth, Owen, Propst, Ashlee, Mehta, Aakash, Nguyen, Kenny H., Kuhlman, Elizabeth, Guseman, Alex J., Stadmiller, Samantha S., Boothby, Thomas C., Neher, Saskia B., Pielak, Gary J.
Format: Article
Language:English
Subjects:
Animals
biologics
Charge density
Deactivation
Dehydrogenase
Dehydrogenases
Desiccation
desiccation tolerance
enzyme protection
Enzyme Stability
Enzymes
Excipients
Formulations
Freeze drying
Freezing
Full‐Length Papers
intrinsically disordered proteins
L-Lactate dehydrogenase
L-Lactate Dehydrogenase - chemistry
L-Lactate Dehydrogenase - metabolism
Lactate dehydrogenase
Lactic acid
Lipase
Lipoprotein lipase
Lipoprotein Lipase - chemistry
Lipoprotein Lipase - metabolism
Low temperature
Models, Molecular
Protein Conformation
Proteins
Proteins - metabolism
Tardigrada - metabolism
tardigrades
Temperature requirements
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Summary:Protein‐based biological drugs and many industrial enzymes are unstable, making them prohibitively expensive. Some can be stabilized by formulation with excipients, but most still require low temperature storage. In search of new, more robust excipients, we turned to the tardigrade, a microscopic animal that synthesizes cytosolic abundant heat soluble (CAHS) proteins to protect its cellular components during desiccation. We find that CAHS proteins protect the test enzymes lactate dehydrogenase and lipoprotein lipase against desiccation‐, freezing‐, and lyophilization‐induced deactivation. Our data also show that a variety of globular and disordered protein controls, with no known link to desiccation tolerance, protect our test enzymes. Protection of lactate dehydrogenase correlates, albeit imperfectly, with the charge density of the protein additive, suggesting an approach to tune protection by modifying charge. Our results support the potential use of CAHS proteins as stabilizing excipients in formulations and suggest that other proteins may have similar potential.
ISSN:0961-8368
1469-896X
DOI:10.1002/pro.3604