Back to Search
Start Over
Fourier transform infrared spectroscopy analysis of the conformational quality of recombinant proteins within inclusion bodies
- Source :
- Biotechnology journal. 3(2)
- Publication Year :
- 2008
-
Abstract
- The solubility of recombinant proteins produced in bacterial cells is considered a key issue in biotechnology as most overexpressed polypeptides undergo aggregation in inclusion bodies, from which they have to be recovered by solubilization and refolding procedures. Physiological and molecular strategies have been implemented to revert or at least to control aggregation but they often meet only partial success and have to be optimized case by case. Recent studies have shown that proteins embedded in inclusion bodies may retain residual structure and biological function and question the former axiom that solubility and activity are necessarily coupled. This allows for a switch in the goals from obtaining soluble products to controlling the conformational quality of aggregated proteins. Central to this approach is the availability of analytical methods to monitor protein structure within inclusion bodies. We describe here the use of Fourier transform infrared spectroscopy for the structural analysis of inclusion bodies both purified from cells and in vivo. Examples are reported concerning the study of kinetics of aggregation and structure of aggregates as a function of expression levels, temperature and co-expression of chaperones.
- Subjects :
- Protein Folding
Protein Conformation
Kinetics
FIS/07 - FISICA APPLICATA (A BENI CULTURALI, AMBIENTALI, BIOLOGIA E MEDICINA)
Applied Microbiology and Biotechnology
Inclusion bodies
law.invention
Protein structure
law
Spectroscopy, Fourier Transform Infrared
Fourier transform infrared spectroscopy
Solubility
Fourier transform infrared spectroscopy · Inclusion bodies · Recombinant proteins · Solubility
Inclusion Bodies
Chemistry
General Medicine
BIO/10 - BIOCHIMICA
Recombinant Proteins
FIS/01 - FISICA SPERIMENTALE
Biochemistry
Solubilization
Recombinant DNA
Molecular Medicine
Electrophoresis, Polyacrylamide Gel
Function (biology)
Subjects
Details
- ISSN :
- 18607314
- Volume :
- 3
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Biotechnology journal
- Accession number :
- edsair.doi.dedup.....fbb5412a7a839c7b838fc75b888a68c2