PMID: 9434273Jan 22, 1998Paper

Real-time in situ monitoring of lysozyme during lyophilization using infrared spectroscopy: dehydration stress in the presence of sucrose

Pharmaceutical Research
R L RemmeleJ F Carpenter

Abstract

First, to investigate the role of sucrose in stabilizing protein structure (as measured by changes in the amide I band of lysozyme) caused by dehydration encountered during lyophilization. Second, to demonstrate the utility of internal reflection spectroscopy as a tool for conducting controlled lyophilization experiments. A custom-built internal reflection FTIR accessory was used to follow the entire freeze-drying process of solutions consisting of 49.4 mg/mL lysozyme in the presence and absence of 10% sucrose in real-time. Studies were carried out using D2O as a transparent medium in the infrared region of the protein amide bands. Potential self-association of the protein in the presence of sucrose was investigated using dynamic light scattering. Hydration levels were determined using a multiple regression equation. Differential scanning calorimetry (DSC) permitted characterization of the final lyophilized product. Moisture content was determined using Karl Fischer titration. Throughout freezing and drying, minimal changes were observed both in frequency (1647 +/- 1 cm-1) and bandwidth (46 +/- 1 cm-1) of the amide I band in the presence of sucrose. In contrast, greater changes in frequency and band width were seen in the absen...Continue Reading

Citations

Nov 22, 2005·Expert Opinion on Drug Delivery·Marco van de WeertSven Frokjaer
May 9, 2001·The Journal of Pharmacy and Pharmacology·Y H LiaoG P Martin
Mar 16, 2005·Optometry and Vision Science : Official Publication of the American Academy of Optometry·Lakshman N SubbaramanLyndon Jones
Jul 28, 2007·European Biophysics Journal : EBJ·S MagazùM T F Telling
Oct 27, 2007·Pharmaceutical Development and Technology·Bakul S BhatnagarMichael J Pikal
Oct 8, 2005·Expert Review of Proteomics·Mark Cornell Manning
Apr 9, 2020·Frontiers in Cell and Developmental Biology·Khairat Bahgat Youssef El BaradieMark W Hamrick
Jan 6, 2004·Journal of Pharmaceutical Sciences·Jeffrey D MeyerJohn F Carpenter
Dec 5, 2006·International Journal of Pharmaceutics·Fei TianJ Howard Rytting
Nov 4, 2006·Journal of Proteome Research·Peng Wu, David W Grainger
Jul 16, 2011·Advanced Drug Delivery Reviews·Satoshi OhtakeTsutomu Arakawa

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