Para la investigación
Genosphere Biotechnologies’s Protein Production team provides extensive services for protein expression, production and purification processes.
Let us know which protein is of interest to your research and we will produce it for you according to your specifications.
Our recombinant proteins have been successfully used in an wide range of experiments including antibodies development, activity assays, crystallography, growth hormones assays, protein-protein interactions, protein-DNA interactions.
Guaranteed, affordable, ready-to-use proteins in a few weeks
Purity >80% or Purity >90%
Standard project workflow
1• Sequence design
A number of studies have revealed that codon usage bias may cause severe expression problems. The presence of rarely used codons for the host translation machinery often leads to :
-Reduced translation rate of the target mRNA.
-Amino acids misincorporation.
-Truncated or amino-acids deleted polypeptides or proteins.
The overall result being low or undetectable protein production.
To help solve the issues related to codon usage bias, the target DNA sequence may be re-defined through codon changes using the appropriate usage frequency table for the host organism: that’s codon optimisation. This general approach has been verified to significantly improve heterologous protein expression in numerous cases.
Project design process includes adding N-/C- terminus TAGS for purification and detection purposes.
2• Gene synthesis and characterisation
• The DNA is synthesised starting with oligonucleotides.
• The synthetic ORF is built and cloned into our standard pUC-derived high copy number plasmid.
• Transformed colonies are screened, a first set of five positive clones are selected, amplified and purified.
• DNA sequence is confirmed
3• Subcloning into an expression vector
• Plasmid prep, restriction digest and gel purification of target fragment, ligation of fragment into the best suited vector(s) for your host.
• Transformation of cells, picking, growing under selective pressure, DNA mini prep.
• DNA sequence confirmation.
• Plasmid Midi/Maxi prep for the confirmed cloned vector.
4• Protein expression assay and optimisation
• Small-scale transfection and reactor flasks to determine the gene’s expression level.
• SDS-PAGE analysis of expression products under various tested conditions.
• Scale-up expression.
5• Scale-up protein purification and characterisation
• Standard TAG-based affinity chromatography column followed by ion exchange and/or gel filtration chromatography.
• Refolding experiment where necessary.
• Concentration determination.
• SDS-PAGE and WESTERN BLOT experiments.
Scale up fermentation and purification
Where to start
You may contact us by email to: firstname.lastname@example.org
Please include the following information:
• Your gene or protein sequence or UniProt accession number
• Your preferred expression system(s)
• The mg amount of recombinant protein needed
• The purification level required: >80% or >90%
► Five expression systems
•Expression in bacteria Escherichia coli
•Expression in yeast Pichia pastoris
•Expression in baculovirus-infected insect cells
•Expression in HEK or CHO cells
•Expression in vitro (E. coli enzymatic transcription and translation machinery)
► Standard protein purity levels
Choice of purification levels required:
Purity >80 % or Purity > 90 %
► Affinity-purification tags
N- or C-terminal
Glutathione S-transferase fusion (GST tag)
6x or 10x histidine residues ((His)6 or (His10)tag)