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Guidelines, Strategies, and Principles for the Directed Evolution of Cross-Reactive Antibodies Using Yeast Surface Display Technology

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Yeast Surface Display

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2491))

Abstract

The ability of cross-reactive antibodies to bind multiple related or unrelated targets derived from different species provides not only superior therapeutic efficacy but also a better assessment of treatment toxicity, thereby facilitating the transition from preclinical models to human clinical studies. This chapter provides some guidelines for the directed evolution of cross-reactive antibodies using yeast surface display technology. Cross-reactive antibodies are initially isolated from a naïve library by combining highly avid magnetic bead separations followed by multiple cycles of flow cytometry sorting. Once initial cross-reactive clones are identified, sequential rounds of mutagenesis and two-pressure selection strategies are applied to engineer cross-reactive antibodies with improved affinity and yet retained or superior cross-reactivity.

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Acknowledgments

We gratefully acknowledge all group members for helpful discussions and critical reading of this chapter. The financial contributions from the iCARE-2 Marie Curie fellowship cofounded by the AIRC foundation for cancer research and the European Union’s Horizon 2020 research and innovation programme to E.L.W. (grant agreement n. 800924) is gratefully acknowledged.

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Author notes

  1. Monica Chinellato

    Present address: Department of Medicine (DIMED), University of Padua, Padua, Italy

Authors and Affiliations

  1. Department of Molecular Sciences and Nanosystems, Ca’ Foscari University of Venice, Mestre, Italy

    Sara Linciano, Ee Lin Wong, Ylenia Mazzocato, Monica Chinellato, Tiziano Scaravetti, Alberto Caregnato, Veronica Cacco, Zhanna Romanyuk & Alessandro Angelini

  2. European Centre for Living Technology (ECLT), Ca’ Bottacin, Venice, Italy

    Alessandro Angelini

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  1. Sara Linciano
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Correspondence to Alessandro Angelini .

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Editors and Affiliations

  1. Department of Chemistry, Institute of Biochemistry, University of Natural Resources and Life Sciences, Vienna, Austria

    Michael W. Traxlmayr

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Linciano, S. et al. (2022). Guidelines, Strategies, and Principles for the Directed Evolution of Cross-Reactive Antibodies Using Yeast Surface Display Technology. In: Traxlmayr, M.W. (eds) Yeast Surface Display. Methods in Molecular Biology, vol 2491. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2285-8_14

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  • DOI: https://doi.org/10.1007/978-1-0716-2285-8_14

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2284-1

  • Online ISBN: 978-1-0716-2285-8

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