The generation and selection of single-domain, v region libraries from nurse sharks.
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.
Flajnik
Martin F
MF
Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
Dooley
Helen
H
D000595:Amino Acid Sequence; D000818:Animals; D017931:DNA Primers / Q000737:chemistry; D007114:Immunization; D007132:Immunoglobulin Isotypes / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000276:immunology*; D007136:Immunoglobulins / Q000276:immunology*; D008969:Molecular Sequence Data; D019151:Peptide Library; D011946:Receptors, Antigen / Q000276:immunology*; D017386:Sequence Homology, Amino Acid; D012754:Sharks / Q000276:immunology*
D017931:DNA Primers; D007132:Immunoglobulin Isotypes; D007135:Immunoglobulin Variable Region; D007136:Immunoglobulins; D019151:Peptide Library; D011946:Receptors, Antigen; C092388:nurse shark antigen receptor