Document Detail


Crystallization of recombinant human growth hormone at elevated pressures: pressure effects on PEG-induced volume exclusion interactions.
MedLine Citation:
PMID:  20589675     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Crystallization of recombinant human growth hormone (rhGH) at elevated pressures was investigated in the presence of 6,000 molecular weight poly(ethylene glycol; PEG-6000). Crystallization of rhGH at atmospheric pressure occurred at a protein concentration of 15 mg/mL in 6% PEG-6000. Crystallization did not occur in the same solutions at 250 MPa. In contrast, at a pressure of 250 MPa in the presence of 8% PEG-6000, rhGH readily crystallized from solutions containing 35 mg/mL rhGH, whereas amorphous precipitate formed in the same solutions at atmospheric pressure. Osmotic virial coefficients were determined from static light scattering measurements and combined with a hard-sphere activity coefficient model to predict rhGH activity coefficients as a function of pressure and PEG concentration. Predicted activity coefficients quantitatively matched those determined from equilibrium solubility measurements. The ability to adjust the thermodynamic non-ideality with pressure provides a valuable tool to study protein crystallization in addition to providing a methodology for obtaining crystals at elevated pressures.
Authors:
Ryan L Crisman; Theodore W Randolph
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  107     ISSN:  1097-0290     ISO Abbreviation:  Biotechnol. Bioeng.     Publication Date:  2010 Nov 
Date Detail:
Created Date:  2010-09-27     Completed Date:  2011-01-06     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  United States    
Other Details:
Languages:  eng     Pagination:  663-72     Citation Subset:  IM    
Copyright Information:
© 2010 Wiley Periodicals, Inc.
Affiliation:
Department of Chemical and Biological Engineering, University of Colorado Center for Pharmaceutical Biotechnology, University of Colorado, Boulder, CO 30309, USA.
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MeSH Terms
Descriptor/Qualifier:
Crystallization / methods
Human Growth Hormone / chemistry*,  metabolism
Humans
Hydrostatic Pressure*
Polyethylene Glycols / chemistry,  metabolism
Recombinant Proteins / chemistry,  metabolism
Technology, Pharmaceutical / methods*
Chemical
Reg. No./Substance:
0/Polyethylene Glycols; 0/Recombinant Proteins; 12629-01-5/Human Growth Hormone

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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