Gene to protein
E. coli, Pichia pastoris
  • Clone the gene of interest into chosen expression vector.
  • Small-scale expression screening. Large-scale (≥1 L) expression culture. Primary protein purification.
Molecular Cloning and Mutagenesis
  • Clone the gene of interest into chosen expression vector. DNA sequence verification.
  • Single or multiple nucleotide base change e.g. amino acid substitution, Insert / delete restriction site, stop codons and small tag. DNA sequence verification.
Plasmid production
  • Production and purification of plasmid DNA from mini- to mega-prep scale.
Small-scale Expression Screening
E. coli
  • Expression screen under multiple conditions. Each construct tested in two different expression strains, two temperatures and two media and two inducer concentrations.
  • Analyze protein expression and affinity binding by SDS-PAGE.
  • Resin-binding assay by affinity chromatography. Determine optimal expression conditions.
Pichia pastoris
  • Expression screen of 10 yeast clones per construct. Analyze expression and affinity binding by SDS-PAGE.
Large-scale Protein Expression and Purification
E. coli or Pichia pastoris
  • Expression at ≥ 1 L in shake flasks. Cells and/or supernatant collected at optimal time of harvest. SDS-PAGE analysis.
  • If required, we can produce Isotope labelled protein for NMR or mass spectrometry applications.
Primary purification
  • Sample extraction followed by affinity chromatography (e.g. IMAC, GST, MBP, Strep(ll)). SDS-PAGE analysis.
Secondary purification
  • Secondary purification to improve protein purity by techniques such as affinity, ion exchange, hydrophobic interaction or size exclusion chromatography. SDS-PAGE analysis.
Removal of Fusion tag
  • Screen for optimal protease cleavage condition to remove fusion tag from protein. Chromatography to recover untagged protein. SDS-PAGE analysis.