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Custom recombinant protein expression and purification in mammalian cells
Protein expression services proposed by Proteogenix are based on a step by step protocol:
Step 0: Gene synthesis and codons optimization
This gene synthesis step is very important because it allows us to optimize rare codons for mammalian cells expression systems and therefore significantly increases in some cases production yields.
Step I: cDNA cloning into an expression vector
This step consists in integrating the cDNA into a mammalian cells expression vector containing a tag and inducing the expression of the fusion protein of interest and allowing the protein purification on affinity column via the tag. It is possible to cleave the tag.
Step II: Transient expression and purification evaluation of recombinant protein
- Transfection of cells (HK293, CHO) with the expression vector
- Evaluation of the expression by Western blot and optimization
- Pilot expression followed by a pilot purification step
Step III: Recombinant protein production and purification scale-up:
- Production run in bioreactor, harvesting and clarification
- Affinity chromatography purification step
- Quality control by acrylamide gel (SDS-PAGE) and / or Western blot