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The Generation and Selection of Single-Domain, V Region Libraries from Nurse Sharks

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 562))

Summary

The cartilaginous fish (sharks, skates, and rays) are the oldest phylogenetic group in which a human-type adaptive immune system and immunoglobulins (Igs) have been found. In addition to their conventional (heavy-light chain heterodimeric) isotypes, IgM and IgW, sharks produce the novel isotype, IgNAR, a heavy chain homodimer that does not associate with light chains. Instead, its variable (V) regions act as independent, soluble units in order to bind antigen. In this chapter, we detail our immunization protocol in order to raise a humoral IgNAR response in the nurse shark (Ginglymostoma cirratum) and the subsequent cloning of the single-domain V regions from this isotype in order to select antigen-specific binders by phage display.

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Correspondence to Helen Dooley .

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Flajnik, M.F., Dooley, H. (2009). The Generation and Selection of Single-Domain, V Region Libraries from Nurse Sharks. In: Aitken, R. (eds) Antibody Phage Display. Methods in Molecular Biology, vol 562. Humana Press. https://doi.org/10.1007/978-1-60327-302-2_6

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  • DOI: https://doi.org/10.1007/978-1-60327-302-2_6

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60327-301-5

  • Online ISBN: 978-1-60327-302-2

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