Creative Biostructure offers customized Mempro™ cell-free protein production services for immunoglobulin-like superfamily (E set) proteins. You can count on us all through your project.
Mempro™ cell-free protein production system is the innovative systems to produce a variety of membrane proteins, which can bypass the limitations that encountered in cell-based protein production system, such as host cell toxicity, low yield and solubilization and purification using detergents. The immunoglobulin superfamily (IgSF) is a group of membrane proteins that play important roles in the recognition, binding, or adhesion processes of cells. Proteins of the IgSF have an Ig domian (also named Ig-like domain), which possesses an Ig-like fold that composed of approximately 70-100 amino acids in anti-parallel β-strands. Immunoglobulin-like superfamily termed E set, includes a wide range of domains that have an Ig-like fold. This superfamily is composed of PKD domains, cadherins and several families of bacterial Ig-like domains as well as viral tail fibre proteins. E set also includes several fibronectin type III domain-containing families.
Figure 1. Ig-like domains in Escherichia coli oxidoreductases. Ribbon diagram of the Ig-like periplasmic SOD SodC. (FEMS Microbiol. Rev., 2013)
Creative Biostructure is capable of overexpressing, purifying and characterizing immunoglobulin-like superfamily (E set) proteins, we can provide various strategies for membrane proteins of interest, including:
Among Mempro™ cell-free protein production platform, E. coli S30 extract is widely used for E set proteins production. Wheat germ extract can provide eukaryotic translation environment for E set protein expression. Baculovirus expression system from Autographa californica nucleopolyhedrovirus (AcNPV) is also a major tool for E set protein production, which is developed in the early stage that cultivated insect cells in combination with vectors derived from the baculovirus. Crude rabbit reticulocyte lysate is also the most widely used for E set protein production due to eukaryotic cell expression system. To adopt an appropriate source for membrane protein expression, many factors, such as the availability of materials, protein origin and complexity, downstream processing, etc., should be considered.
One challenge associated with Mempro™ cell-free protein production is that degradation of the DNA by endogenous nucleases. Creative Biostructure can optimize this expression system using plasmids owing to they are circular and have no end for the exonucleases to attach.
We can provide other various Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.
References:Cell-free protein synthesis. (https://en.wikipedia.org/wiki/Cell-free_protein_synthesis)F. Bernhard, et al. (2013). Cell-free expression - making a mark. Cur. Opin. Struct. Biol., 23: 374-380.G. Bodelón, et al. (2013). Immunoglobulin domains in Escherichia coli and other enterobacteria: from pathogenesis to applications in antibody technologies. FEMS Microbiol. Rev., 37(2): 204-250.Ig-like fold superfamily (E-set). EMBL-EBI. (http://pfam.xfam.org/clan/CL0159)Immunoglobulin superfamily. (https://en.wikipedia.org/wiki/Immunoglobulin_superfamily).