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231224s2013 xx |||||o 00| ||eng c |
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|a 10.1021/la4028223
|2 doi
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|a pubmed24n0775.xml
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|e rakwb
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| 041 |
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|a eng
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| 100 |
1 |
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|a Wiesbauer, Johanna
|e verfasserin
|4 aut
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| 245 |
1 |
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|a Renewal of the air-water interface as a critical system parameter of protein stability
|b aggregation of the human growth hormone and its prevention by surface-active compounds
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|c 2013
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|a Text
|b txt
|2 rdacontent
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|a ƒaComputermedien
|b c
|2 rdamedia
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|a ƒa Online-Ressource
|b cr
|2 rdacarrier
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|a Date Completed 24.07.2014
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|a Date Revised 10.12.2013
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|a published: Print-Electronic
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|a Citation Status MEDLINE
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|a Soluble proteins are often highly unstable under mixing conditions that involve dynamic contacting between the main liquid phase and a gas phase. The recombinant human growth hormone (rhGH) was recently shown to undergo aggregation into micrometer-sized solid particles composed of non-native (mis- or unfolded) protein, once its solutions were stirred or shaken to generate a continuously renewed air-water interface. To gain deepened understanding and improved quantification of the air-water interface effect on rhGH stability, we analyzed the protein's aggregation rate (r(agg)) at controlled specific air-water surface areas (a(G/L)) established by stirring or bubble aeration. We show that in spite of comparable time-averaged values for a(G/L) (≈ 100 m(2)/m(3)), aeration gave a 40-fold higher r(agg) than stirring. The enhanced r(agg) under aeration was ascribed to faster macroscopic regeneration of free a(G/L) during aeration as compared to stirring. We also show that r(agg) was independent of the rhGH concentration in the range 0.67 - 6.7 mg/mL, and that it increased linearly dependent on the available a(G/L). The nonionic surfactant Pluronic F-68, added in 1.6-fold molar excess over rhGH present, resulted in complete suppression of r(agg). Foam formation was not a factor influencing r(agg). Using analysis by circular dichroism spectroscopy and small-angle X-ray scattering, we show that in the presence of Pluronic F-68 under both stirring and aeration, the soluble protein retained its original fold, featuring native-like relative composition of secondary structural elements. We further provide evidence that the efficacy of Pluronic F-68 resulted from direct, probably hydrophobic protein-surfactant interactions that prevented rhGH from becoming attached to the air-water interface. Surface-induced aggregation of rhGH is suggested to involve desorption of non-native protein from the air-water interface as the key limiting step. Proteins or protein aggregates released back into the bulk liquid appear to be essentially insoluble
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|a Journal Article
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|a Research Support, Non-U.S. Gov't
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|a Surface-Active Agents
|2 NLM
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|a Water
|2 NLM
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|a 059QF0KO0R
|2 NLM
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|a Poloxamer
|2 NLM
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| 650 |
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|a 106392-12-5
|2 NLM
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| 650 |
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|a Human Growth Hormone
|2 NLM
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| 650 |
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|a 12629-01-5
|2 NLM
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| 700 |
1 |
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|a Prassl, Ruth
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Nidetzky, Bernd
|e verfasserin
|4 aut
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| 773 |
0 |
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|i Enthalten in
|t Langmuir : the ACS journal of surfaces and colloids
|d 1992
|g 29(2013), 49 vom: 10. Dez., Seite 15240-50
|w (DE-627)NLM098181009
|x 1520-5827
|7 nnns
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| 773 |
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|g volume:29
|g year:2013
|g number:49
|g day:10
|g month:12
|g pages:15240-50
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|u http://dx.doi.org/10.1021/la4028223
|3 Volltext
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