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Specific volume and adiabatic compressibility measurements of native and aggregated recombinant human interleukin‐1 receptor antagonist: Density differences enable pressure‐modulated refolding
Author(s) -
Seefeldt Matthew B.,
Crouch Chris,
Kendrick Brent,
Randolph Theodore W.
Publication year - 2007
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.21398
Subject(s) - adiabatic process , compressibility , chemistry , hydrostatic pressure , volume (thermodynamics) , bar (unit) , thermodynamics , partial specific volume , yield (engineering) , antagonist , hydrostatic equilibrium , interleukin 1 receptor antagonist , recombinant dna , receptor , receptor antagonist , biochemistry , physics , quantum mechanics , meteorology , gene , enzyme
High hydrostatic pressures have been used to dissociate non‐native protein aggregates and foster refolding to the native conformation. In this study, partial specific volume and adiabatic compressibility measurements were used to examine the volumetric contributions to pressure‐modulated refolding. The thermodynamics of pressure‐modulated refolding from non‐native aggregates of recombinant human interleukin‐1 receptor antagonist (IL‐1ra) were determined by partial specific volume and adiabatic compressibility measurements. Aggregates of IL‐1ra formed at elevated temperatures (55°C) were found to be less dense than native IL‐1ra and refolded at 31°C under 1,500 bar pressure with a yield of 57%. Partial specific adiabatic compressibility measurements suggest that the formation of solvent‐free cavities within the interior of IL‐1ra aggregates cause the apparent increase in specific volume. Dense, pressure‐stable aggregates could be formed at 2,000 bar which could not be refolded with additional high pressure treatment, demonstrating that aggregate formation conditions and structure dictate pressure‐modulated refolding yields. Biotechnol. Bioeng. 2007;98; 476–485. © 2007 Wiley Periodicals, Inc.