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Crystallization of recombinant human growth hormone at elevated pressures: Pressure effects on PEG‐induced volume exclusion interactions

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Abstract:

Abstract

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. Biotechnol. Bioeng. 2010;107:663–672. © 2010 Wiley Periodicals, Inc.

Document Type: Research Article

DOI: http://dx.doi.org/10.1002/bit.22832

Affiliations: Department of Chemical and Biological Engineering, University of Colorado Center for Pharmaceutical Biotechnology, University of Colorado, Boulder, Colorado 30309; telephone: 303-492-4776;, Fax: 303-492-4341

Publication date: November 1, 2010

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