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Crystallization of recombinant human growth hormone at elevated pressures: Pressure effects on PEG‐induced volume exclusion interactions
Author(s) -
Crisman Ryan L.,
Randolph Theodore W.
Publication year - 2010
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.22832
Subject(s) - crystallization , solubility , virial coefficient , peg ratio , osmotic pressure , chemistry , volume (thermodynamics) , amorphous solid , thermodynamics , ethylene glycol , chromatography , atmospheric pressure , human growth hormone , crystallography , biochemistry , hormone , organic chemistry , growth hormone , economics , physics , oceanography , finance , geology
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.