Protein Identification
What is your desired application? For example, viral or bacterial antigens for diagnostic applications, enzymes, etc. Bioinformatics & simulated 3D protein modelling – The biophysical characteristics will influence the choice of expression system used; which in turn affects the gene design.
Gene Design & Optimisation
The genetic code is designed with the necessary tags, signal peptides, etc. Triplet codon usage is also optimised depending on the choice of expression vector. For custom protein design, in order to accelerate development cycles, more than one gene will be synthesised and cloned to allow for flexibility in the development process.
Sub-cloning, Mapping & Sequencing
Following synthesis of the gene of interest it is sub-cloned into the selected expression vector. Subsequent screening using various methods facilitate the selection of the desired clones, which are then ready to be inserted into the selected expression system for protein production.
Cell Transformation & Expression
Our protein expression platforms include bacteria (E.coli) and mammalian cells (HEK293). With our proprietary technology, we are able to deliver highly stable and functionally active recombinant proteins.
Pilot Scale Production
In order to check whether the protein of interest is indeed being synthesized a pilot scale production platform is set up. In a relatively short period of time we can assess whether the desired protein is being synthesised in a functionally active form.
Analytical protein characterization ensures the identity, purity, structural and conformational integrity and activity the protein. A number of protein analyses are routinely performed during all project stages. These include electrophoresis (SDS-PAGE, native-page, western blot, ELISA (direct or sandwich), protein assay (A280, BCA or equivalent)
Purification
The purification steps adopted will be determined not only by the expression system used but also the characteristics of the synthesised protein.
Our proprietary purification technologies facilitate the folding of the protein in order to ensure it retains its native functional characteristics.
SDS-PAGE and western blotting are routinely used for fraction analysis, while final protein purity is determined by densitometry from Coomassie and or Silver-stained gels.
We have also validated purity via Mass Spectrometry. This highly sensitive analytical technique measures the mass-to-charge ratio of ions. The results displayed as a mass spectrum, plots intensity as a function of the mass-to-charge ratio, and demonstrates our extremely high levels of protein purity.
Stability Studies
One of the biggest hurdles to overcome in successful recombinant protein production is ensuring that the protein of interest is packaged in a functionally active and stable form – our first prize is to ship proteins in a lyophilised form, at ambient temperature.
The stability of a protein is influenced by a number of things. The buffer conditions affect the overall surface charge of the proteins, their tendency to form aggregates or precipitate out of solution, as well as their capacity to be lyophilised in native form. Our proprietary buffer formulations do not denature our proteins, enable facile reconstitution and facilitate easy gold conjugation.
Optimisation of Expression
We aim to achieve maximum recombinant protein production by optimization of fermentation processes.
Our experience with various high-producing cell lines, cell culture growth conditions, media formulations, methods of protein extraction and enrichment enables us to develop the most optimal production process for protein of interest.
Commercial Scale Production
Our array of fermentation systems and bioreactors allow fully flexible scale-up protein production, aggressive delivery timescales and very competitive pricing.
Delivery
Products are supplied with a certificate of analysis, purification report and a batch record provided at specified protein concentration. They are lyophilised if feasible, or formulated in a buffer that protects from protein degradation or precipitation dispensed into specified aliquot sizes.