Molecular biology:
- Choice of production plasmid
- Tag used and its position in the sequence
- Sequence optimization to avoid rare codons
![Plasmide_opt](http://p2r-protein-facility.eu/wp-content/uploads/2018/11/Plasmide_opt-300x300.png)
![Structure_opt](http://p2r-protein-facility.eu/wp-content/uploads/2018/11/Structure_opt.png)
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
![NGC_Bio-rad_opt](http://p2r-protein-facility.eu/wp-content/uploads/2018/05/NGC_Bio-rad_opt-200x196.jpg)
![Limulus_opt](http://p2r-protein-facility.eu/wp-content/uploads/2018/11/Limule_opt.png)
Endotoxins:
- Titration with lysate of limulus amoebocytes (LAL) kit
- Elimination by affinity column