Services
NMR-Bio has the expertise and technologies available to answer your challenges and support you throughout your full project.
Provide us with the sequence of your gene and we will deliver you the protein with the required structural study by recommending and implementing the best strategy. NMR-Bio uses both cell-based and cell-free systems to provide customized protein expression services to customers in the pharmaceutical industry, as well as research institutions.
I- Protein sample production
NMR-Bio have developed relevant labelling protocols in various expression systems including:
Each system has its advantages and liabilities. The selection of the system depends on the type, required amount and functional activities of the protein. The experts at NMR-Bio will choose the appropriate expression system to obtain high quality proteins to fulfil your specific research goals aiming at high resolution structural solution and innovative drug discovery.
E. coli: E. coli is the easiest expression system to perform isotopic labelling for simple proteins. Our scientists are expert in deuteration and methyl specific labelling of large proteins. NMR-Bio manufactures patented precursors for the production of (13C, 1H)-labelled proteins and uses them for the custom production of your protein of interest. Precursors are introduced at a specific time in a perdeuterated minimal medium to achieve high incorporation levels of 13CH3 isotopomer only in a single type of methyl group without detectable scrambling. We also perform combinatorial labelling with protocols designed to label simultaneously different types of methyl probes well separated in Methyl-TROSY spectra.
We also routinely conduct 13C, 15N, 2H uniform labelling in flasks or fermenters.
Please contact us at service@nmr-bio.com for your specific needs.
Yeast: Yeasts has many of the advantages of higher eukaryotic expression systems such as protein processing, protein folding and post-translational modification. Protocols for isotopic labelling are well described and give high incorporation rates.
We have a project to develop efficient methyl specific isotopic labelling in yeast (Pichia pastoris) in 2020.
If you are interested, please contact us at: service@nmr-bio.com to set up a collaborative project together.
Insect cell: Baculovirus expression systems offer high-level expression, ease of scale-up and production of recombinant proteins with posttranslational modifications. NMR-Bio has developed a culture medium and advanced protocols for the specific labelling of methyl groups of your protein or protein complex. We utilize optimized and fully validated insect cell expression protocols for consistent reproducible high-level protein expression in serum-free shake flasks.
Mammalian cells: NMR-Bio is currently developing stable mammalian cell lines including non-human mammalian cell lines (CHO) and human cell lines (HEK293) for recombinant protein expression. These expression systems are able to produce post-translational modifications which closely mimic those in human. Various biotherapeutics including antibodies, enzymes, cytokines or hormones are produced by non-human mammalian expression systems by pharmaceuticals industries.
Cell-free: In vitro protein synthesis or cell-free protein synthesis (CFPS) is an approach for protein production without living cells, but with cell extracts containing the necessary molecular machinery. This makes it ideal solution for expressing difficult-to-produce proteins, such as toxins or membrane proteins…
NMR-Bio’s cell extracts are made from E. coli lysate. In particular, we have developed improved extracts for your disulphide bridge containing proteins (e.g. FAB, mAbs...) and we have access to a dedicated cell-free platform to ensure maximum yields of functional proteins.
NMR-Bio offers 15N, 13C, 2H uniform labelling without scrambling in the Cα position of attached protons or deuterium. This eliminates the back-exchange problem and enables full assignment of the backbone of your protein of interest. Moreover, NMR-bio’s innovative proprietary cell-free technology is tailored for methyl specific labelling of proteins in a perdeuterated background.
We provide synthesis in batch for conditions screening and in dialysis mode up to 100 mL of reaction for large-scale production of your labelled samples.
Please contact us at service@nmr-bio.com for your specific needs.
II- Structural studies:
Efficient and reliable NMR structural studies require acquisition of high quality experimental data and specialist knowledge for interpretation. Our senior scientists with extensive experience offer NMR services including but not limited to :
Quality control (e.g. refolding, compactness, structure modification);
Molecular structure characterization;
Structure determination;
NMR spectra assignment;
Reaction kinetics investigation;
Dynamics, disorder analysis;
Quantitative analysis;
Drug screening;
Mapping structure of biological drugs by NMR;
NMR analysis for biopharmaceutical development;
High field NMR data acquisition.
For more information about our services or to receive a specific quotation corresponding to your project, please contact us by email at: service@nmr-bio.com.
Our NMR sample preparation guidelines:
Isotopic labelling: protein NMR spectroscopy requires isotope enrichment. NMR-Bio offer uniform and specific isotopic labelling of your protein sample in various expression systems.
Amount:for solution NMR, a sample typically contains 1-10 mg/mL of 0,3-0,5 mM purified protein in a suitable buffer.
Stability:For in-depth studies by NMR, the protein needs to be stable for several days at room temperature for data acquisition.
Size:the molecular weight of biomacromolecules of a NMR sample is typically less than 50 kDa for high-resolution structure determination. NMR-Bio is an expert in methyl specific labelling for the characterization of larger proteins and protein complexes.
High field NMR Equipment:
We have access to state-of-the-art NMR instrumentation like Bruker AVANCE 900 MHz, AVANCE 850 MHz with sample changer, AVANCE 700 MHz and other spectrometers all equipped with sophisticated cryoprobes including the adapted for small volumes. With these modern NMR equipment, we can offer our specialized services to industrial customers.
