Abstract
Antibody libraries came into existence 25 years ago when the accumulating sequence data of immunoglobulin genes and the advent of the PCR technology made it possible to clone antibody gene repertoires. Phage display (most common) and additional display and screening technologies were applied to pan out desired binding specificities from antibody libraries. “Synthetic” or “semisynthetic” libraries are from naive—non-immunized source and considered to be a source for many different targets, including self-antigens.
As other antibody discovery tools, phage display is not an off-the-shelf technology and not offered as a kit but rather requires experience and expertise for making it indeed very useful. Here we present application notes that expand the usefulness of antibody phage display as a very versatile and robust antibody discovery tool.
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Acknowledgments
In 2009, we published in “Methods in Molecular Biology” a chapter describing the construction of a large human synthetic single-chain Fv antibody library displayed on phage, where in vivo formed complementarity determining regions (CDRs) were shuffled combinatorially onto germline-derived human variable region frameworks [15]. The present chapter is a revision and update of that chapter, not including the part of library construction. We are grateful to past and present members of the Benhar Lab for their contributions in optimizing the antibody phage display protocols described herein.
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Bitton, A., Nahary, L., Benhar, I. (2018). Antibody Isolation From a Human Synthetic Combinatorial and Other Libraries of Single-Chain Antibodies. In: Hust, M., Lim, T. (eds) Phage Display. Methods in Molecular Biology, vol 1701. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7447-4_19
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DOI: https://doi.org/10.1007/978-1-4939-7447-4_19
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