The generation and selection of single-domain, v region libraries from nurse sharks

Martin F Flajnik, Helen Dooley

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

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.
Original languageEnglish
Pages (from-to)71-82
Number of pages12
JournalMethods in Molecular Biology
Volume562
DOIs
Publication statusPublished - 2009

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Keywords

  • cartilaginous fish
  • shark
  • immunoglobin
  • IgNAR
  • variable region
  • single domain

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