The SCOPE® technology allows us to generate E. coli strains expressing high yields of the target protein and tight control of the expression of the protein.
SCOPE® technology comprises a set of expression vectors that can be used in combination with our GMP-compliant E. coli strains or any other preferred, customer-derived, E. coli strain.
The SCOPE® vectors are modular build-expression vectors allowing rapid exchange of diverse elements such as promoters, origin of replication, leader sequences, antibiotics resistance markers, and gene of interest. Each of these vectors is mapped, enabling rapid design and construction of custom-made plasmids, tailored to individual needs. Our ready-to-go SCOPE® vectors have proven value in providing tight control of protein expression in cases where the protein of interest interferes with biomass formation, or simply in high yield production of proteins in inclusion bodies, soluble format, or secreted to the periplasmic fraction. Using our SCOPE® plasmid series, we have demonstrated very high yields of proteins, for example 5 g/L of green fluorescent protein (GFP).
We typically deploy our SCOPE® protein expression technology to deliver high quality research material in short timelines (from DNA to positive clones in approximately 4-5 weeks) and at reduced cost. Proteins can be produced in inclusion bodies, solubly or secreted into the periplasmic space, upon induction with a sugar. The induced expression typically ranges from a couple of hours to a couple of days and can be scaled-up rapidly and cost efficiently by using animal component-free media and other raw materials. First the DNA is synthesized and used to transform competent E. coli cells. We typically select around 10 positive clones that are subsequently cultured in small scale systems, such as our SCOUT® system, to demonstrate proper protein expression. Protein expression and functionality is usually demonstrated by SDS-PAGE, western blot analyses and/or activity assays. Based on such data sets, the best three clones are selected, and research cell banks are prepared. All procedures and documents are GMP compliant, so that generated research clones can be seamlessly transitioned into a GMP trajectory. Next, we assess optimal biomass formation in stirred tank bioreactors or shake flasks and assess optimal protein production parameters not only for yield, but also from a quality perspective. Such experiments are typically performed at a 250 mL to 2 L scale. The SCOPE® technology thus delivers stable protein expressing E. coli banks, protocols for robust protein production, and research-grade protein material for pre-clinical studies. The materials and documentation we deliver facilitate a direct path into scale-up and clinical manufacturing.