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Custom recombinant protein expression and purification in baculovirus
Protein expression service proposed by Proteogenix is 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 baculovirus expression system 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 baculovirus 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: Recombinant virus generation and recombinant protein expression evaluation in baculovirus
- Generation of recombinant bacmid DNA
- Transfection of insect cells with recombinant bacmid
- Generation of virus stocks
- Infection of insect cells
- Quality control by acrylamide gel (SDS-PAGE) and / or Western blot
Step III: Recombinant protein production and purification from 1 Liter:
- Protein expression and purification from one or more liters of insect cell culture
- Affinity purification run on column
- Quality control by acrylamide gel (SDS-PAGE) and / or Western blot
Turnaround time :
- Cloning : 2 weeks
- Expression evaluation : 5 to 6 weeks
- Production and purification : 1 to 2 weeks