Enzyme Dehydration Using Microglassification™ Preserves the Protein's Structure and Function. Issue 2 (29th December 2014)

• Record Type: Journal Article
• Authors: Aniket; Gaul, David A.; Bitterfield, Deborah L.; Su, Jonathan T.; Li, Victoria M.; Singh, Ishita; Morton, Jackson; Needham, David
• Is Part Of: Journal of pharmaceutical sciences. Volume 104:Issue 2(2015:Feb.)
• Page Start: 640
• Page End: 651
• Publication Date: 2014-12-29
• Subjects: Pharmacy -- Periodicals; 615.1
• DOI: 10.1002/jps.24279
• Languages: English
• ISSNs: 0022-3549

Abstract

<abstract abstract-type="main" xml:lang="en"> <title> <x xml:space="preserve">Abstract</x> </title> <p>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 <italic>n</italic>‐pentanol, <italic>n</italic>‐octanol, <italic>n</italic>‐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 > <italic>n</italic>‐pentanol > triacetin > <italic>n</italic>‐octanol > <italic>n</italic>‐decanol. Enzymes Microglassified™ in <italic>n</italic>‐pentanol retained higher activity (93%–98%) than <italic>n</italic>‐octanol (78%–85%) or <italic>n</italic>‐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 (<italic>p</italic> < 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. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 104:640–651, 2015</p> </abstract>