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dc.contributor.authorQuaas, Bastian
dc.contributor.authorBurmeister, Laura
dc.contributor.authorLi, Zhaopeng
dc.contributor.authorSatalov, Alexandra
dc.contributor.authorBehrens, Peter
dc.contributor.authorHoffmann, Andrea
dc.contributor.authorRinas, Ursula
dc.date.accessioned2020-01-29T10:03:44Z
dc.date.available2020-01-29T10:03:44Z
dc.date.issued2019-11-20
dc.identifier.citationPharm Res. 2019 Nov 20;36(12):184. doi: 10.1007/s11095-019-2705-5.en_US
dc.identifier.issn1573-904X
dc.identifier.pmid31748894
dc.identifier.doi10.1007/s11095-019-2705-5
dc.identifier.urihttp://hdl.handle.net/10033/622106
dc.description.abstractPURPOSE: There is a plethora of studies on recombinant human bone morphogenetic protein-2 (rhBMP-2) application and delivery systems, but surprisingly few reports address the biophysical properties of the protein which are of crucial importance to develop effective delivery systems or to solve general problems related to rhBMP-2 production, purification, analysis and application. METHODS:The solubility, stability and bioactivity of rhBMP-2 obtained by renaturation of E. coli derived inclusion bodies was assessed at different pH and in different buffer systems using (dynamic) light scattering and thermal shift assays as well as intrinsic fluorescence measurements and luciferase based bioassays. RESULTS: rhBMP-2 is poorly soluble at physiological pH and higher. The presence of divalent anions further decreases the solubility even under acidic conditions. Thermal stability analyses revealed that rhBMP-2 precipitates are more stable compared to the soluble protein. Moreover, correctly folded rhBMP-2 is also bioactive as precipitated protein and precipitates readily dissolve under appropriate buffer conditions. Once properly formed rhBMP-2 also retains biological activity after temporary exposure to high concentrations of chaotropic denaturants. However, care should be taken to discriminate bioactive rhBMP-2 precipitates from misfolded rhBMP-2 aggregates, e.g. resolvability in MES buffer (pH 5) and a discrete peak in thermoshift experiments are mandatory for correctly folded rhBMP-2. CONCLUSIONS: Our analysis revealed that E. coli derived rhBMP-2 precipitates are not only bioactive but are also more stable compared to the soluble dimeric molecules. Knowledge about these unusual properties will be helpful to design improved delivery systems requiring lower amounts of rhBMP-2 in clinical applications.en_US
dc.language.isoenen_US
dc.publisherSpringeren_US
dc.rightsAttribution-NonCommercial-ShareAlike 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.subjectprotein aggregationen_US
dc.subjectprotein solubilityen_US
dc.subjectprotein stabilityen_US
dc.subjectrecombinant human bone morphogenetic protein-2en_US
dc.subjectrefoldingen_US
dc.titleStability and Biological Activity of E. coli Derived Soluble and Precipitated Bone Morphogenetic Protein-2.en_US
dc.typeArticleen_US
dc.contributor.departmentHZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.en_US
dc.identifier.journalPharmaceutical Researchen_US
dc.source.journaltitlePharmaceutical research


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