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Mempro™ Cell-Free Immunoglobulin Superfamily Protein Production

Based on the comprehensive protein engineering platform established through years of experiences, scientists form Creative Biostructure can provide advanced costum Mempro™ protein production services for immunoglobulin superfamily (IgSF) proteins using cell-free expression system. You can count on us all through your project.

Mempro™ cell-free protein production system is the innovative system to produce integral membrane proteins, which bypasses the limitations exsited in cell-based protein production system, such as host cell toxicity, low yield and solubilization and purification using detergents. IgSF is a large group of proteins that are involved in the recognition, binding, or adhesion processes of cells, including CD4, CD8, T cell receptor (TCR), and HLA. IgSF proteins have a structural domain termed as immunoglobulin domain (Ig domain), which is about 70-110 amino acids in length. Ig domain has a sandwich-like structure formed by two sheets of antiparallel beta strands. The Ig domain can be classified as IgV, IgC1, IgC2, and IgI.


Figure 1. The immunoglobulin superfamily. (Wikipedia)

Creative Biostructure has extensive expertise in high-yield IgSF protein production based on cell-free protein expression system, we can provide various strategies for Mempro™ cell-free protein production, including:

Among Mempro™ cell-free membrane protein production systems, E. coli S30 extract is widely used for IgSF protein production. Wheat germ extract can provide eukaryotic translation system for IgSF protein expression. Baculovirus expression system from Autographa californica nucleopolyhedrovirus (AcNPV) is also a major system for IgSF protein expression. Crude rabbit reticulocyte lysate is another powerful tool for eukaryotic cell-free protein expression. To select a suitable source of the extract, many factors, such as the availability of materials, protein origin and complexity, downstream processing, etc., should be considered.

With the Mempro™ cell-free protein production platform, Creative Biostructure are capable of expressing, purifying and crystallizing IgSF proteins to perform further structural and functional studies. Mempro™ cell-free protein production has a wide range of advantages, including:

  • Very high yield;
  • Low cost in time and money;
  • Lack of toxicity;
  • Incorporation of non-natural amino acids into protein structure;
  • Relatively tolerant of additives.

One challenge associated with cell-free protein production is that degradation of the DNA by endogenous nucleases. Creative Biostructure can optimize Mempro™ cell-free protein production for IgSF proteins using plasmids because of they are circular and have no end for the exonucleases to attach.

We provide other various Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.

References:
F. Bernhard, et al. (2013). Cell-free expression - making a mark. Cur. Opin. Struct. Biol., 23: 374-380.
Immunoglobulin superfamily. (https://en.wikipedia.org/wiki/Immunoglobulin_superfamily)
K. Natarajan, et al. (2015). Immunoglobulin superfamily. eLS, doi: 10.1002/9780470015902.a0000926.pub2


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