TARGET EXPRESSION
NMX offers different expression platforms for target production, including E. coli for recombinant proteins and in-vitro production systems for proteins and RNA. The combination of these technologies allows our scientists to tailor expression workflows to our client needs, whether the requirement is a high-yield protein production for different downstream assays or a rapid screening of challenging or toxic proteins.
Why NMX?
At NMX, we know that producing a high-quality target is often the major bottleneck in different downstream applications in particular for challenging targets. NMX combines flexibility, experience, and scientific precision to support your project.
E. coli protein expression systems deliver high yields at low cost through the use of optimized strains, customized vector design, and profound expertise in overcoming protein expression challenges — such as weak expression, low protein solubility, and intrinsically disorder proteins (IDPs). On the other hand, a cell-free protein expression system is favored when tight timelines or protein toxicity are limiting factors, since it enables rapid production without the need for living cells.
Our protein production experts are targeting workflows to meet the demands of advanced downstream applications, including biophysical and structural studies. With fast turnaround, in-depth quality control, and robust customization options, we make sure projects are handled to satisfy your needs.
E. coli Expression System
Fast, scalable and reliable
E. coli expression is a widely used system to produce recombinant proteins that provides rapid growth with possible genetic manipulation at low cost. We work with various E. coli strains and custom vector designs to improve solubility and maximize product yield.
Our expert team leverages their knowledge to tackle different challenges such as low yield, poor solubility, and hard-to-express proteins, to ultimately maximize success rates.
NMX offers small-scale expression screening before scaling up to optimize expression conditions and reduce the risk of unsatisfactory production yield. The expression test includes examining multiple E. coli strains, various induction temperatures, and distinct IPTG concentrations to identify the best conditions for the target protein, which is critical for proteins that lack a well-established protocol.
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Throughout the years, NMX has gained significant expertise in production of isotopically labeled proteins, with advanced labeling strategies including 15N, 13C, 19F, 2H, and selective residue labeling such as ILV(A) which enables a variety of NMR spectroscopy techniques to study binding interactions, conformational changes, and dynamics.
In-vitro target production systems
Speed and flexibility
In-vitro target production systems enable scientists to produce biomolecules without the need to rely on living cells. These systems exploit the cellular machinery, such as ribosomes and enzymes, to synthesize biological macromolecules in a controlled environment.
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In-vitro transcription (IVT) is now a widely used method for producing RNA-based biological therapeutics. Starting from a dsDNA template, the desired ssRNA is generated by the T7RNA polymerase. Reaction conditions can be optimized and scaled up to produce mg quantities of RNA in just hours. Moreover, this method allows for custom sequence design and the incorporation of modified or isotopically labeled nucleotides.
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Cell-free protein synthesis (CFPS), also known as in-vitro translation, is a relatively new technology for protein production. This system depends on cell extracts to synthesize proteins in a controlled in-vitro environment. This technology provides efficient and rapid protein production, avoiding the complexities and limitations of in-vivo expression. As such, it is well-suited for proteins generating cell toxicity or requiring presence of additives to fold correctly (i.e. membrane proteins), and it offers high throughput capabilities for applications such as protein engineering. Furthermore, it allows custom and specific labeling options (15N, 13C, 2H), incorporation of unnatural amino acids with easy protein production monitoring. Although this type of protein expression system is generally used in small-scale, it is a scalable technique when it’s worth the investment.
