Protein and solutes freeze-concentration in water/glycerol mixtures revealed by pulse EPR
DC Element | Wert | Sprache |
---|---|---|
dc.contributor.author | Isaev, Nikolay | |
dc.contributor.author | Steinhoff, Heinz-Jurgen | |
dc.date.accessioned | 2021-12-23T16:10:03Z | - |
dc.date.available | 2021-12-23T16:10:03Z | - |
dc.date.issued | 2021 | |
dc.identifier.issn | 09396411 | |
dc.identifier.uri | https://osnascholar.ub.uni-osnabrueck.de/handle/unios/9133 | - |
dc.description.abstract | Lyophilization can extend protein drugs stability and shelf life, but it also can lead to protein degradation in some cases. The development of safe freeze-drying approaches for sensitive proteins requires a better understanding of lyophilization on the molecular level. The evaluation of the freezing process and its impact on the protein environment in the nm scale is challenging because feasible experimental methods are scarce. In the present work we apply pulse EPR as a tool to study the local concentrations of the solute in the 20 nm range and of the solvent in the 1 nm range for a spin labeled 27 kDa monomeric green fluorescent protein, mEGFP, and the 172 Da TEMPOL spin probe, frozen in different water/glycerol-d5 mixtures. For average glycerol volume fractions, phi >= 0.4 we observed transparent glassy media; the local concentration and the 1 nm solvent shell of TEMPOL and the protein correspond to those of a uniform vitrified glass. At phi <= 0.3 we observed partial ice crystallization, which led to ice exclusion of glycerol and TEMPOL with freeze-concentration up to the glycerol maximal-freeze local volume fraction, phi, of 0.64. The protein concentration and its shell behavior was similar except for the lowest phi (0.1), which showed a 4.7-fold freeze-concentration factor compared to sevenfold for TEMPOL, and also a smaller phi. We explain this behavior with an increased probability for proteins to get stuck in the ice phase during fast freezing at higher freeze-concentration and the related large-scale mass transfer. | |
dc.description.sponsorship | Alexander von Humboldt FoundationAlexander von Humboldt Foundation; Russian Federal Ministry of Science and Higher Education; We thank Prof. Dr. Jacob Piehler for providing mEGFP, Drs. Svetlana Kucher and Anna Matveeva for discussions. This work was supported by Alexander von Humboldt Foundation and core funding from the Russian Federal Ministry of Science and Higher Education. | |
dc.language.iso | en | |
dc.publisher | ELSEVIER | |
dc.relation.ispartof | EUROPEAN JOURNAL OF PHARMACEUTICS AND BIOPHARMACEUTICS | |
dc.subject | AGGREGATION | |
dc.subject | DEER | |
dc.subject | DISTANCE MEASUREMENTS | |
dc.subject | ESEEM | |
dc.subject | Freeze-concentration | |
dc.subject | Freezing | |
dc.subject | Local concentration | |
dc.subject | Lyophilization | |
dc.subject | MEMBRANE | |
dc.subject | Pharmacology & Pharmacy | |
dc.subject | Protein nearest shell | |
dc.subject | SOLID-STATE | |
dc.subject | SPIN | |
dc.subject | STABILIZATION | |
dc.subject | TRICHOGIN GA-IV | |
dc.subject | WATER CONCENTRATION | |
dc.title | Protein and solutes freeze-concentration in water/glycerol mixtures revealed by pulse EPR | |
dc.type | journal article | |
dc.identifier.doi | 10.1016/j.ejpb.2021.08.017 | |
dc.identifier.isi | ISI:000702827300006 | |
dc.description.volume | 169 | |
dc.description.startpage | 44 | |
dc.description.endpage | 51 | |
dc.identifier.eissn | 18733441 | |
dc.publisher.place | RADARWEG 29, 1043 NX AMSTERDAM, NETHERLANDS | |
dcterms.isPartOf.abbreviation | Eur. J. Pharm. Biopharm. | |
crisitem.author.dept | Universität Osnabrück | - |
crisitem.author.dept | FB 04 - Physik | - |
crisitem.author.deptid | fb04 | - |
crisitem.author.parentorg | Universität Osnabrück | - |
crisitem.author.netid | IsNi001 | - |
crisitem.author.netid | StHe633 | - |
Seitenaufrufe
3
Letzte Woche
1
1
Letzter Monat
1
1
geprüft am 01.06.2024