Abstract
Ribosome display is a cell-free technology which enables in vitro selection and evolution of antibodies from very large diversified DNA libraries. It operates through the following key steps: (1) generation of PCR library; (2) formation of stable antibody-ribosome-mRNA (ARM) complexes as the selection particles, (3) selection of ligand-binding ARM complexes on an immobilized ligand and (4) recovery of the selected genetic information as DNA by RT-PCR. Since PCR-based random or/and site-directed mutagenesis can be easily used to introduce mutations into the selected DNA pool in each cycle, ribosome display offers an efficient “protein evolution” tool for antibody optimization. Both prokaryotic and eukaryotic cell-free systems have been explored for ribosome display of different proteins. In this chapter, we describe the use of the eukaryotic rabbit reticulocyte ribosome display method to isolate variants of VH antibody fragments with improved affinities.
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References
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Acknowledgments
Research at Crescendo Biologics is funded by Sofinnova Partners, Avlar BioVentures, Aitua Limited and the Rainbow Seed Fund. The Babraham Institute is supported by Biotechnology and Biological Sciences Research Council (BBSRC), UK.
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Edwards, B.M., He, M. (2012). Evolution of Antibodies In Vitro by Ribosome Display. In: Chames, P. (eds) Antibody Engineering. Methods in Molecular Biology, vol 907. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-974-7_16
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DOI: https://doi.org/10.1007/978-1-61779-974-7_16
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Publisher Name: Humana Press, Totowa, NJ
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Online ISBN: 978-1-61779-974-7
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