Generation and isolation of target-specific single-domain antibodies from shark immune repertoires

Methods Mol Biol. 2012;907:177-94. doi: 10.1007/978-1-61779-974-7_9.

Abstract

The drive to exploit novel targets and biological pathways has lead to the expansion of classical antibody research into innovative fragment adaptations and novel scaffolds. The hope being that alternative or cryptic epitopes may be targeted, tissue inaccessibility may be overcome, and easier engineering options will facilitate multivalent, multi-targeting approaches. To this end, we have been isolating shark single domains to gain a greater understanding of their potential as therapeutic agents. Their unique shape, small size, inherent stability, and simple molecular architecture make them attractive candidates from a drug discovery perspective. Here we describe protocols to capture the immune repertoire of an immunized shark species and to build and select via phage-display target-specific IgNAR variable domains (VNARs).

MeSH terms

  • Animals
  • Bacteriophages / metabolism
  • Cell Surface Display Techniques
  • DNA, Complementary / biosynthesis
  • Epitopes / immunology
  • Immunization
  • Molecular Biology / methods*
  • Protein Structure, Tertiary
  • RNA / blood
  • Sharks / immunology*
  • Single-Domain Antibodies / blood
  • Single-Domain Antibodies / chemistry
  • Single-Domain Antibodies / genetics*
  • Single-Domain Antibodies / isolation & purification*

Substances

  • DNA, Complementary
  • Epitopes
  • Single-Domain Antibodies
  • RNA