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mRNA display is an in vitro selection and evolution technique for functional peptides and proteins. In mRNA display, in vitro translation enables the production of proteins using cellular machinery in a cell-free system. These proteins can then undergo subsequent screening for desired properties. CD BioSciences offers mRNA display-based drug discovery services that encompass the entire early drug discovery process.
Fig 1. General scheme for peptide selection by mRNA display.
In vitro translation, also termed in vitro protein expression, cell-free protein synthesis, or cell-free translation, allows rapid production of small functional protein quantities. Cell-free protein synthesis systems mimic cellular transcription and translation processes in a controlled extracellular environment, enabling detailed exploration of individual components and reaction networks.
Fig 2. A comparison of cell-free and in vivo protein synthesis methods. (Gregorio, et al., 2019)
With the rapid advancement of cell-free protein synthesis (CFPS), a variety of CFPS systems have emerged. One category is the extract-based systems, which can be subcategorized into high-utilization cell types and low-utilization cell types based on the source of the cell extract. Another category is the PURE system (Protein Synthesis Using Recombinant Elements), developed by Shimizu et al., which synthesizes proteins using recombinant elements in a cell-free transcription-translation system.
| Translation Systems | Description | Advantages | Disadvantages |
| Rabbit Reticulocyte Lysate | Derived from rabbit reticulocytes, this system is rich in ribosomes and translation initiation factors. |
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| Wheat Germ Extract | Extracted from wheat germ, it's particularly effective for plant-derived proteins. |
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| E. coli Extract | E. coli cell-free systems consist of a crude extract that is rich in endogenous mRNA. |
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| Insect Cell-Free Systems | Obtained from insect cells, these systems provide translation efficiencies similar to mammalian cells. |
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| Mammalian Cell-Free Systems | Derived from mammalian cells, offering translation and post-translational modifications akin to those occurring in vivo. |
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| PURE (protein synthesis using recombinant elements) | PURE system utilizes individually purified components in place of cell extract. |
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In mRNA display, commonly employed translation systems include the rabbit reticulocyte lysate and the PURE system. CD BioSciences utilizes the PURE system for in vitro translation processes. With the PURE system, all components are well-defined, devoid of nucleases and proteases, allowing for the optimization of each element's concentration to achieve maximal protein expression. These reorganization elements include the following.
CD BioSciences' in vitro translation process can rapidly generate substantial amounts of protein for further characterization, saving valuable laboratory time and making it ideal for high-throughput technologies.
These recombinant components are mixed with ribosomes, tRNA, essential NTPs and amino acids isolated from E. coli, ATP, and recombinant T7 RNA polymerase to synthesize proteins using mRNA-puromycin conjugates.
CD BioSciences is committed to providing expert guidance and support throughout the drug development process. We work closely with our clients to develop tailored strategies that maximize the potential of their drug development programs. To learn more about how we can support your drug development efforts with our mRNA display services, please contact us.
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