What we do with your amino acid sequence:
- Optimize the sequence
- Synthesize the gene
- Clone the gene into a vector
- Produce large-scale plasmid DNA
- Optimize protein expression
- Bacterial Protein Production
- Tagged or untagged versions
- Variety of vectors
- Solubility enhancing fusions
- Purification method development
- Mammalian Protein Production
- Protein production in HEK293 or CHO cells
- Protein production using stable transient pools
- Purification using FPLC
- SDS Page
- Western Blot
- Analytical SEC
- Mass Spectrometry
- Polishing SEC
- Endotoxin Level Measurement
- 15N Isotope labeling for protein-NMR
Case Studies for Protein Production
1. Production of THIOMAB in Mammalian Cells
The received THIOMAB DNA sequence was optimized for codon usage, the gene synthesized and cloned it into a vector under a strong CMV promoter and a strong secretory sequence. The cells were transiently transfected and several clones were tested for protein expression. Protein expression was optimized using different conditions, such as temperature and different H:L chain ratios.
The produced THIOMAB protein was purified by Protein A affinity chromatography. Our transient transfection yielded the maximum amount of protein, which was further analyzed for purity and aggregation by SEC.
2. Production of Beta-Galactosidase in E. coli Bacteria
Our client provided the amino acid sequence of cold-active beta-Galactosidase. The sequence was optimized for codon usage in E. coli. The gene was then synthesized and cloned into a bacterial vector and protein production was optimized to maximize expression and solubility.
The protein was purified using Ni+ NTA protocols and was then subjected to functional and activity assays. After a final polishing step by SEC, the exact mass of beta-galactosidase was confirmed by mass spectrometry (TOF).