caa a22 4500 477127177 CHVBK 20180405111653.0 cr unu---uuuuu 170330e19971001xx s 000 0 eng 10.1023/A:1012112503590 doi (NATIONALLICENCE)springer-10.1023/A:1012112503590 Long-Term and High-Temperature Storage of Supercritically-Processed Microparticulate Protein Powders [Elektronische Daten] [Michael Winters, Pablo Debenedetti, Jannette Carey, H. Sparks, Samir Sane, Todd Przybycien] Purpose. The long-term and high-temperature storage of dry, micron-sized particles of lysozyme, trypsin, and insulin was investigated. Subsequent to using supercritical carbon dioxide as an antisolvent to induce their precipitation from a dimethylsulfoxide solution, protein microparticles were stored in sealed containers at -25, -15, 0, 3, 20, 22, and 60°C. The purpose of this study was to investigate the suitability of supercritical antisolvent precipitation as a finishing step in protein processing. Methods. Karl Fisher titrations were used to determine the residual moisture content of commercial and supercritically-processed protein powders. The secondary structure of the dry protein particles was determined periodically during storage using Raman spectroscopy. The proteins were also redissolved periodically in aqueous buffers and assayed spectrophotometrically for biological activity and by circular dichroism for structural conformation in solution. Results. Amide I band Raman spectra indicate that the secondary structure of the protein particles, while perturbed from that of the solution state, remained constant in time, regardless of the storage temperature. The recoverable biological activity upon reconstitution for the supercritically-processed lysozyme and trypsin microparticles was also preserved and found to be independent of storage temperature. Far UV circular dichroism spectra support the bioactivity assays and further suggest that adverse structural changes, with potential to hinder renaturation upon redissolution, do not take place during storage. Conclusions. The present study suggests that protein precipitation using supercritical fluids may yield particles suitable for long-term storage at ambient conditions. Plenum Publishing Corporation, 1997 supercritical fluid antisolvent nationallicence protein precipitation nationallicence secondary structure nationallicence solid state nationallicence long-term storage nationallicence Winters Michael Department of Chemical Engineering, Princeton University, 08544-5263, Princeton, New Jersey aut Debenedetti Pablo Department of Chemical Engineering, Princeton University, 08544-5263, Princeton, New Jersey aut Carey Jannette Chemistry Department, Princeton University, 08544-5263, Princeton, New Jersey aut Sparks H. Isermann Department of Chemical Engineering, Rensselaer Polytechnic Institute, 12180-3590, Troy, New York aut Sane Samir Isermann Department of Chemical Engineering, Rensselaer Polytechnic Institute, 12180-3590, Troy, New York aut Przybycien Todd Isermann Department of Chemical Engineering, Rensselaer Polytechnic Institute, 12180-3590, Troy, New York aut Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/10(1997-10-01), 1370-1378 0724-8741 14:10<1370 1997 14 11095 https://doi.org/10.1023/A:1012112503590 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1012112503590 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Winters Michael Department of Chemical Engineering, Princeton University, 08544-5263, Princeton, New Jersey aut NATIONALLICENCE 700 1- Debenedetti Pablo Department of Chemical Engineering, Princeton University, 08544-5263, Princeton, New Jersey aut NATIONALLICENCE 700 1- Carey Jannette Chemistry Department, Princeton University, 08544-5263, Princeton, New Jersey aut NATIONALLICENCE 700 1- Sparks H. Isermann Department of Chemical Engineering, Rensselaer Polytechnic Institute, 12180-3590, Troy, New York aut NATIONALLICENCE 700 1- Sane Samir Isermann Department of Chemical Engineering, Rensselaer Polytechnic Institute, 12180-3590, Troy, New York aut NATIONALLICENCE 700 1- Przybycien Todd Isermann Department of Chemical Engineering, Rensselaer Polytechnic Institute, 12180-3590, Troy, New York aut NATIONALLICENCE 773 0- Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/10(1997-10-01), 1370-1378 0724-8741 14:10<1370 1997 14 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer