Case Studies2018-02-15T12:32:44+00:00

Production and analysis of an extensively post-translationally modified secreted protein

We were asked to generate a secreted protein that is known to undergo significant post-translational modification. The challenge was to not only make it but to clearly demonstrate that all the required post-translational modifications had occurred. A HEK293 system was chosen because, as a mammalian cell line, this was most likely to generate the required modifications. The protein was secreted using an efficient, artificial signal [...]


Production of a post-translationally modified protein

Our client required 20mg of protein for NMR structural studies. The protein was known to be post-translationally modified with palmitic acid on a cysteine residue. Our challenge was to obtain this amount of protein after expression in E. coli and demonstrate palmitoylation using mass spectrometry analysis. The protein was well expressed but very insoluble in initial experiments. Our Solutions [...]


Generation of a novel, secreted protein for structure determination

Our client required >2 mg of a novel, secreted protein for X-ray structure determination studies. The protein was thought to be N-glycosylated. Our challenge was to obtain sufficient >95% pure protein for crystallisation experiments, as measured by SDS-PAGE, size exclusion and mass spectrometry analysis.  In addition, we were also aiming to take forward the shortest, most stable construct feasible to maximise our chances [...]