PMID: 15368989Sep 17, 2004Paper

The application and mechanisms of polyethylene glycol 8000 on stabilizing lactate dehydrogenase during lyophilization

PDA Journal of Pharmaceutical Science and Technology
Yanli Mi, George Wood

Abstract

The purpose of this paper is to explore the application and mechanisms of polyethylene glycol 8000 (PEG 8000) on stabilizing lactate dehydrogenase (LDH) during lyophilization. In earlier freeze-thawing experiments, different molecular weights and concentrations of PEGs were formulated with LDH, and ultraviolet (UV) enzymatic activity and circular dichroism (CD) wavelength scanning studies were conducted. In lyophilization studies, different molecular weights of saccharides, e.g., glucose, sucrose, dextran 37,000 (D 37K), and dextran 160,000 (D 160K), with or without PEG 8000, were formulated with LDH at various molar ratios. UV assays, size exclusion chromatography -high performance liquid chromatography (SEC-HPLC), CD, fourier transform infrared spectroscopy (FTIR) were conducted for LDH. Upon lyophilization, enzymatic activity and tetrameric structure recoveries of LDH-saccharide formulations reached over 90% with PEG 8000 vs. 60-80% without PEG 8000. LDH-PEG 8000-saccharide formulations shifted the melting temperature (Tm) to higher temperatures than did LDH-saccharide formulations. Most LDH-PEG 8000-saccharide formulations at 1:100:1000 molar ratio showed better preservation of LDH secondary structures than did LDH-sacchari...Continue Reading

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