Premium
High‐pressure refolding of human vascular endothelial growth factor (VEGF) recombinantly expressed in bacterial inclusion bodies: Refolding optimization, and feasibility assessment
Author(s) -
Cothran Amber,
John Richard J. St.,
Schmelzer Charles H.,
Pizarro Shelly A.
Publication year - 2011
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.642
Subject(s) - sparging , inclusion bodies , hydrostatic pressure , chemistry , yield (engineering) , recombinant dna , high pressure , urea , chemical engineering , process engineering , chromatography , biochemistry , materials science , organic chemistry , composite material , physics , engineering physics , engineering , gene , thermodynamics
High‐pressure has been established as an effective technique for refolding proteins at high concentrations. In this study, high hydrostatic pressure (1–3 kbar) was utilized to refold a homodimeric protein from inclusion bodies and the process was evaluated for large‐scale manufacturing feasibility. This research focused on increasing protein concentration while maximizing yield and product quality. Refolding yields of 29–42% were achieved in the absence of urea at 2 kbar and at a protein concentration of 6 g/L. Optimization of the refolding buffer composition via multivariate design of experiments and other process parameters such as refolding pressure, gas sparging, and time under pressure are discussed. Although high‐pressure refolding can be considered a viable technology for manufacturing if the gains are clearly identified, in this particular case, the benefits that the high‐pressure technology offers do not compensate for the drawbacks of implementing new equipment in an existing facility, and unknown impact of scale‐up for this molecule. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2011