All West Bioscience customized protein expression and purification are carried out in our California facility to insure highest quality and best customer satisfaction. We have been a leader in providing high quality recombinant protein expression and purification services, especially in customized protein design area. We delivered thousands proteins to our customers worldwide.
- Potentially very high expression levels
- Low cost
- Rapid growth
- Simple transformation protocols
- Many parameters can be altered to optimize expression
- Inefficient disulfide bond formation
- Poor folding of proteins in the cytoplasm
- Inclusion body formation
- In vitro refolding protocols inefficient – may negate advantages
- Codon usage different to eukaryotes
- Minimal post-translational modifications
WE will clone your insert into an expression vector by either a standard subcloning or a PCR cloning. will be used to transfer the gene of interest into the expression vector. Clones will be fully sequenced for verification.
The expression vector containing the gene of interest will be transformed into an expression strain of E. coli for protein production. Several individual clones will be screened in a small-scale induction experiment. The expression results will be analyzed by SDS/PAGE and Western blot.
The expression condition will be optimized by time of induction, temperature and media condition. The results will be confirmed by SDS-PAGE and Western blot.
A small-scale culture will be tested and the expressed protein will be characterized for cellular location, solubility, yield and accessibility of purification tags. A detailed strategy for producing and purifying the protein will be designed.
Large-scale production in liters volume will be made using shake flasks. If you prefer purifying protein by your own, you will receive the cell pellets and a complete report with the SDS-PAGE and Western blot analysis results.
Several methods can be used to purify your protein such as inclusion body preparation or affinity column purification.