Protein expression and purification

THE PROTEIN EXPRESSION AND PURIFICATION CENTRE

The Protein expression and purification centre provides expertise, molecular tools and equipment for the expression of recombinant proteins in prokaryotic and eukaryotic systems and for the purification of proteins from all sources.

Protein expression

The workflow for recombinant protein production comprises 4 main stages:

Experimental design
Cloning
Small scale expression screening
Scale-up

and comprises:

prokaryotic / eukaryotic systems
main expression hosts are bacteria and insect cells (Baculovirus vectors)
other expression hosts are yeast and mammalian cells
production of Single proteins
production of Protein complexes (co expression)
Refolding
Production of Affinity purified polyclonal antibodies

Protein Purification

The workflow for protein purification comprises 5 main stages:

Experimental design
cell lysis / extraction
Purification
Functional assay
Quality control

and comprises:

Classic Biochemistry (e.g. precipitation, dif. centrifugation)
Ultrafiltration for concentration and buffer exchange
Chromatography:
- Preparative
- High resolution polishing purification
- Analytical (High resolution)
- @ Room temperature and 4ºC
- Size Exclusion (SEC)
- F-SEC (fluorescence detection in normal phase SEC)
- Ion exchange (IEX)
- Hydrophobic interaction
- Reverse-phase (peptide analysis and purification)
Analytical assays

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