We design, express and purify customised proteins reagents to support all stages of the drug discovery process. Our cell and protein scientists have a wealth of experience, many from large pharma organisations.

Design of the right protein is critical for success.  We use a variety of bioinformatic packages to model and engineer the target protein. This allows us to maximise success for a range of different uses including screening, structural biology, biophysics and in vivo studies. We explore different construct length, different domains, point mutations, affinity tags and sometimes chimeras in order to get the best solution.

We have successfully used HEK293 suspension cells for many years, for both secretory and intracellular protein expression. We routinely use the baculovirus/insect cell system, working with the flashBACTM technology from Oxford Expression Technologies. Both of these, and in particular HEK293 cells, enable the most authentic post-translational modifications for human proteins which is key for any protein expression service.  Secreted proteins are purified from cell culture media.  Our systems use protein free media, greatly simplifying the purification process.

We also frequently use E. coli because of the speed and quantity of protein that can be achieved when a particular protein expresses well. We manipulate the standard T7 based expression vector systems to maximise soluble expression. We can refold proteins from E. coli-derived inclusion bodies.

All the standard affinity chromatographic procedures are used regularly with a variety of different tags. The design and selection of the most appropriate affinity tag can have a significant impact on the purification process and protein yields achieved. In addition, proteins are purified according to size, charge, hydrophobicity and other properties depending on the protein characteristics. A suite of Akta systems are available for the work.

As part of our protein production service, proteins are analysed by a variety of methods. SDS-PAGE is used under both reducing and non-reducing conditions as required, to look for disulphide bonds. Purity and size in solution is determined using size exclusion chromatography. Protein concentration is established using absorbance at 280nm, correcting for the extinction coefficient. Liquid chromatography mass spectrometry is used in two ways.  If the protein will “fly” in the mass spectrometer –the intact mass of the protein is determined to search for post-translational modifications.  The peptide map can be determined after proteolytic digestion. The thermal stability of the protein will also be monitored if needed.

OTHER SERVICES

Protein Crystallography and Structure Determination

Our protein structure determination service is driven by an outstanding team of experienced crystallographers with a strong publication record in solving a wide variety of protein structures, including novel ones. A key part of this process is the design of the protein and the ability to screen many different constructs and crystallisation conditions. Our final delivery package is not just a set of co-ordinates. We supply a detailed analysis of the overall 3D structure, along with our learning and conclusions, with ideas for project progression and drug design. Interactive discussions and regular communication with our clients are a key part of our service.

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Protein Mass spectrometry

We now provide intact protein and peptide mapping as a stand-alone service to anyone requiring detailed protein or antibody characterisation.  Key to this service is the flexibility and quick turnaround times we offer with no detailed contract set-up. Our expert mass spectrometrist, Rachel Rowlinson, will initially discuss your specific needs. Help will be giving with planning and interpretation of the samples analysed and a comprehensive data summary package will be provided with follow-up discussion if required. Whether you have one sample or larger numbers on a regular basis will be reflected in our pricing.

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GET IN TOUCH WITH PEAK PROTEINS

For further information or simply to say hello please contact us on 01625 238892 or email us