Abstract
The selection and concomitant affinity maturation of proteins to bind to user-defined target molecules have become a key technology in biochemical research, diagnostics, and therapy. One of the most potent selection technologies for such applications is ribosome display. It works entirely in vitro, and this has two important consequences. First, since no transformation of any cells is required, libraries with much greater diversity can be handled than with most other techniques. Second, since a library does not have to be cloned and transformed, it is very convenient to introduce random errors in the library by PCR-based methods and select improved binders. Thus, a true directed evolution, an iteration between randomization and selection over several generations, can be conveniently carried out, e.g., for affinity maturation. Ribosome display has been used successfully for the selection of antibody fragments and other binding proteins, such as Designed Ankyrin Repeat Proteins (DARPins).
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Acknowledgments
We thank many former and current members of the Plückthun laboratory for establishing and continuously optimizing the ribosome display protocol. In this respect we thank the following people not only, but especially for the following efforts: Petra Parizek, Birgit Lindner, and Gabriela Nagy-Davidescu for revision of the pRDV plasmid sequence, Oliver Scholz for the testing of the EWT5s primer, amplification conditions, and RNA yield, Fabio Parmeggiani for testing of the illustra G50 columns, and Gautham Varadamsetty and Hilmar Ebersbach for their help in testing enzymes on different protein scaffolds. Work on ribosome display was supported by the Swiss National Science Foundation.
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Dreier, B., Plückthun, A. (2011). Ribosome Display: A Technology for Selecting and Evolving Proteins from Large Libraries. In: Park, D. (eds) PCR Protocols. Methods in Molecular Biology, vol 687. Humana Press. https://doi.org/10.1007/978-1-60761-944-4_21
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DOI: https://doi.org/10.1007/978-1-60761-944-4_21
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