Projects per year
Abstract
Protein-coated microcrystals can be generated by co-precipitation of protein and a water-soluble crystalline carrier by addition to excess water miscible organic solvent. We have investigated this novel process for its utility in the concentration and partial purification of a recombinant protein exported into the culture broth during expression by Pichia pastoris. Co-precipitation with a L-glutamine carrier selectively isolated the protein content of the culture broth, with a minimal number of steps, and simultaneously removed contaminants including a novel yeast metabolite. This pigment co-elutes during aqueous chromatography but its elucidation as a benzoylated glycosamine suggested a simple route of removal by partition during the co-precipitation process. Scale-up of the process was readily achieved through in-line mixing and subsequent reconstitution of the dried protein-coated microcrystals yielded natively folded, bioactive protein. Additional washing of the crystals with saturated L-glutamine facilitated further purification of the recombinant protein immobilized on the L-glutamine carrier. Thus, we present a novel method for the harvesting of recombinant protein from culture broth as a dry powder, which may be of general applicability to bioprocessing.
Original language | English |
---|---|
Pages (from-to) | 764-773 |
Number of pages | 10 |
Journal | Biotechnology and Bioengineering |
Volume | 106 |
Issue number | 5 |
DOIs | |
Publication status | Published - 1 Aug 2010 |
Keywords
- chemical precipitation
- desiccation
- fungal proteins
- glutamine
- pichia
- powders
- recombinant proteins
Fingerprint
Dive into the research topics of 'Isolation of recombinant proteins from culture broth by co-precipitation with an amino acid carrier to form stable dry powders'. Together they form a unique fingerprint.Projects
- 1 Finished
-
New Approaches to High Throughput Protein, Isolation, Purification and Concentration
Moore, B., Cormack, P., Sefcik, J. & Van Der Walle, C.
BBSRC (Biotech & Biological Sciences Research Council)
29/10/07 → 28/06/11
Project: Research