Enzyme dehydration using Microglassification™ preserves the protein's structure and function

[No Value] Aniket, David A Gaul, Deborah L Bitterfield, Jonathan T Su, Victoria M Li, Ishita Singh, Jackson Morton, David Needham

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Abstrakt

Controlled enzyme dehydration using a new processing technique of Microglassification™ has been investigated. Aqueous solution microdroplets of lysozyme, α-chymotrypsin, catalase, and horseradish peroxidase were dehydrated in n-pentanol, n-octanol, n-decanol, triacetin, or butyl lactate, and changes in their structure and function were analyzed upon rehydration. Water solubility and microdroplet dissolution rate in each solvent decreased in the order: butyl lactate > n-pentanol > triacetin > n-octanol > n-decanol. Enzymes Microglassified™ in n-pentanol retained higher activity (93%-98%) than n-octanol (78%-85%) or n-decanol (75%-89%), whereas those Microglassified™ in triacetin (36%-75%) and butyl lactate (48%-79%) retained markedly lower activity. FTIR spectroscopy analyses showed α-helix to β-sheet transformation for all enzymes upon Microglassification™, reflecting a loss of bound water in the dried state; however, the enzymes reverted to native-like conformation upon rehydration. Accelerated stressed-storage tests using Microglassified™ lysozyme showed a significant (p < 0.01) decrease in enzymatic activity from 46,560 ± 2736 to 31,060 ± 4327 units/mg after 3 months of incubation; however, it was comparable to the activity of the lyophilized formulation throughout the test period. These results establish Microglassification™ as a viable technique for enzyme preservation without affecting its structure or function.

OriginalsprogEngelsk
TidsskriftJournal of Pharmaceutical Sciences
Vol/bind104
Udgave nummer2
Sider (fra-til)640-51
ISSN0022-3549
DOI
StatusUdgivet - feb. 2015
Udgivet eksterntJa

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Citationsformater

Aniket, N. V., Gaul, D. A., Bitterfield, D. L., Su, J. T., Li, V. M., Singh, I., Morton, J., & Needham, D. (2015). Enzyme dehydration using Microglassification™ preserves the protein's structure and function. Journal of Pharmaceutical Sciences, 104(2), 640-51. https://doi.org/10.1002/jps.24279