| Author: AG Clark/J Horsfield
Email: alan.clark@vuw.ac.nz
Home Page: http://www.vuw.ac.nz/~clarkag
HPLC
Reverse-phase HPLC
Uses of reverse-phase HPLC
Hydrophobic interaction chromatography (HIC)
Isoelectric focussing and chromatofocussing.
Chromatofocussing
Gel filtration chromatography
PPT Slide
Rationale for gel filtration
Applications of gel filtration
Affinity chromatography
Rationale for affinity chromatography
Affinity chromatography - the matrix.
Affinity chromatography - the ligand.
Ligands:
Overexpression of a protein from a cloned gene
Overexpression of a protein from a cloned gene
Bacterial protein expression
Fusion proteins
Fusion proteins
Recovery of fusion proteins
Eukaryotic overexpression of genes
Affinity recovery of proteins that have been overexpressed
in bacteria:
Method for affinity recovery
Removal of the vector protein…..
….and re-purification of the target protein.
“Tagged” proteins.
Some examples of tags:
Some examples of tags:
Advantages of tagged proteins |