We help you to produce proteins for your research. In our cell culture lab, our staff performs all the work (because of the risk of contaminations in a cell culture lab), but you can help with other parts of the work. This includes vector constructions and analyses of proteins that we have produced, depending on your know-how and experiences. If you want we can perform all the steps. Please contact us to discuss the design of a project for your proteins.
A typical work-flow can be as follows:
1. Vector construction. The cDNA encoding your protein of interest must be cloned into an expression vector for mammalian cells. If you have the possibility, you can perform this step yourself, or we can do it at MPE. We can also help with the ordering of commercially generated vectors.
2. Transfection and cloning. The vector will then be transfected into cells for the generation of stable, producing clones. Analysis of the clones can either be done at MPE or can be done by you.
3. Adaptation. A selected high-producing clone has to be adapted to growth in suspension in serum-free medium prior to a bioreactor can be run for production.
4. Bioreactor run. When a clone is well-adapted, it can be cultured in a bioreactor for production of a larger batch of the recombinant protein. We use perfusion culture where culture supernatant is harvested continuously and we normally collect between 10 and 20 L, depending on the productivity of the clone and how much protein is wanted.
5. Product concentration and purification. The harvested medium is then concentrated by tangential flow filtration, down to volumes that are easier to handle and then purified by for example affinity chromatography.
6. Product analysis. SDS-PAGE, Western blots or ELISAs of the produced protein can be performed throughout the different steps, either by MPE or by you.
As an alternative to this workflow, we can also produce proteins through transient transfection of CHO or HEK293 cells in litre-scale. Steps 2-4 above are then replaced by a larger transfection of many cells, which is performed in our bioreactors. This is a good alternative to get the protein somewhat faster.