Evaluation of Predictors of Protein Relative Stability Obtained by Solid-State Hydrogen/Deuterium Exchange Monitored by FTIR
Autor: | Robin H. Bogner, Michael J. Pikal, Rui Fang, Wasfy M. Obeidat |
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Rok vydání: | 2020 |
Předmět: |
Hydrogen
Size-exclusion chromatography Analytical chemistry Pharmaceutical Science chemistry.chemical_element 02 engineering and technology 030226 pharmacology & pharmacy 03 medical and health sciences chemistry.chemical_compound Freeze-drying 0302 clinical medicine medicine Pharmacology (medical) Pharmacology Chemistry Organic Chemistry 021001 nanoscience & nanotechnology Human serum albumin Trehalose Deuterium Molecular Medicine Hydrogen–deuterium exchange 0210 nano-technology Glass transition Biotechnology medicine.drug |
Zdroj: | Pharmaceutical Research. 37 |
ISSN: | 1573-904X 0724-8741 |
Popis: | Hydrogen/deuterium (H/D) exchange over a range of temperatures suggests a protein structural/mobility transition in the solid state below the system glass transition temperature (Tg). The purpose of this study was to determine whether solid-state protein stability correlates with the difference between storage temperature and apparent Td where an abrupt change in mobility occurs, or alternatively, the extent of H/D exchange at a single temperature correlates directly to protein stability in lyophilized solids. Solid-state H/D exchange was monitored by FTIR spectroscopy to study the extent of exchange and the apparent transition temperature in both pure recombinant human serum albumin (rHSA) and rHSA formulated with sucrose or trehalose. H/D exchange of freeze-dried formulations at 11% RH and temperatures from 30 to 80°C was monitored. Protein stability against aggregation at 40°C/11% RH for 6 months was assessed by size exclusion chromatography (SEC). Both sucrose and trehalose showed equivalent protection of protein secondary structure by FTIR. The rHSA:sucrose formulation showed superior long-term stability at 40°C by SEC over the trehalose formulation, but the apparent Td determined from H/D exchange was much higher in the trehalose formulation. Instead, the extent of H/D exchange (X∞) was lower in the sucrose formulation at the temperature of the stability studies (40°C) than found for the trehalose formulation, which was consistent with better stability in the sucrose formulation. While apparent Td did not correlate with protein stability for rHSA, the extent of H/D exchange, X∞, did. |
Databáze: | OpenAIRE |
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