Recombinant Protein
03/06/2024Recombinant proteins are typically expressed and amplified using E. coli, yeast, and mammalian cells. In recent years, the use of insect and plant cells for recombinant protein production has become increasingly common.
After appropriate pretreatment methods such as clarification, lysis, ultrafiltration, extraction, and centrifugation, recombinant protein fermentation broth yields samples ready for purification. In the subsequent purification process, ion exchange and/or affinity chromatography are often used for crude and intermediate purification. For fine purification, hydrophobic interaction chromatography, size-exclusion chromatography, and multimodal chromatography are commonly employed. Small molecules like insulin and GLP-1 heavily rely on reversed-phase and size-exclusion chromatography resins. To ensure high activity, chiral separation purification is sometimes necessary.
For small molecule recombinant proteins, a fusion protein strategy can be used. This involves appropriate affinity chromatography and enzymatic cleavage to quickly obtain high-purity intermediates, typically meeting purity standards after 1-2 additional chromatography steps. If using the coupling ligand method, re-purification is required after coupling modification to remove mis-modified molecules and unbound monomers.
