Molecular biology:

  • Choice of production plasmid
  • Tag used and its position in the sequence
  • Sequence optimization to avoid rare codons

Protein expression in:

  • Escherichia coli bacteria (BL21(DE3) pLysS, Rosetta, etc):
    • Soluble form
    • Inclusion bodies and renaturation, in particular for MHC/peptide, MR1 and MicA
  • Stable or transient mammalian eukaryotic cells (CHO-K, CHO-S, ExpiCHO) and insects (S2, Sf9)

Purification of proteins or peptides by liquid chromatography:

  • Steric exclusion
  • Ion exchange
  • Hydrophobic interactions
  • Reverse phase
  • Affinity

Endotoxins:

  • Titration with lysate of limulus amoebocytes (LAL) kit
  • Elimination by affinity column