Abstract
Besides classical antibodies with the composition of heavy and light chains, sharks produce a unique heavy chain only isotype, termed Immunoglobulin New Antigen Receptor (IgNAR), in which antigen binding is solely mediated by a single domain, referred to as vNAR. Owing to their high affinity and specificity combined with their small size and high stability, vNAR domains emerged as promising target-binding scaffolds that can be tailor-made for biotechnological and biomedical applications. Herein, we describe protocols for the construction of semi-synthetic, CDR3-randomized vNAR libraries for the isolation of target-specific antibodies using yeast surface display or phage display as platform technology. Additionally, we provide information for affinity maturation of target-specific molecules through CDR1 diversification and sublibrary establishment.
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Acknowledgments
We thank Michael Hust for discussion and advice related to the page-display section of this chapter. Furthermore, we gratefully acknowledge funding from Merck Lab@Technische Universität Darmstadt.
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Grzeschik, J. et al. (2018). Generation of Semi-Synthetic Shark IgNAR Single-Domain Antibody Libraries. 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_8
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DOI: https://doi.org/10.1007/978-1-4939-7447-4_8
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