Protein Factory – crystallographic grade protein production
The protein factory produces crystallographic grade recombinant protein for investigations using the wide range of NovAliX biophysical techniques: NMR, SPR, ITC, and MS. In addition proteins are produced and supplied directly to clients. De novo protein, labeled protein and off-the-shelf protein from the protein gallery are provided with MS, SDS-PAGE and DLS quality control data.
If a target is already in the protein gallery, the protein is freshly prepared to ensure it is of the highest quality and delivered within 2 weeks. The gallery includes a large number of nuclear receptors, kinases and epigenetics targets. Reproduction of literature data is performed with a quick turn-around time.
Years of experience in protein production are the basis of a reliable gene to protein service for de novo targets. The probability of success within the shortest timeframe is achieved with multiple constructs designed after extensive sequence analysis supported by bioinformatics. Codon optimized constructs are tested simultaneously in bacteria and baculovirus infected insect cells. Purification produces proteins of the highest quality (identity, purity, integrity and monodispersity). Proteins can be produced with any fusion tag including; His, GST, MBP, TRX, Strep. Proteins are available in quantities of 5 to 50 mg and can be supplied according to client specifications – concentration, buffer, size of aliquot and package.
NovAliX produces labeled proteins: seleno-methionine labeled protein for de novo structural determination and 15N and 13C labeled protein for NMR structural studies.
Protein quality control – leaving nothing to chance
Proteins produced by NovAliX are rigorously purified and characterized for identity, purity, and homogeneity. The principal tools are sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE), dynamic light scattering (DLS) and mass spectrometry (MS). SDS-PAGE confirms purity and size. DLS measures any aggregation that may be present.
Arguably the most informative technique is MS. MS analysis of whole proteins under denaturing and native conditions will reveal the presence of any adventitious ligands. Other methods would not reveal this potentially vital piece of information.
If greater characterization is required then MS is again particularly powerful. The presence of post-translational modifications such as glycosylation or phosphorylation can be confirmed by their removal by specific chemical or enzymatic reactions. Peptide mapping, i.e. protein cleavage with proteases may be used to confirm the unique identity of the protein or even to confirm the full sequence and most modifications.