Preparation of a naïve library of camelid single domain antibodies

Methods Mol Biol. 2012:911:65-78. doi: 10.1007/978-1-61779-968-6_5.

Abstract

The preparation of antibody libraries starting from lymphocytes recovered from immunized members of the Camelidae enables to collect binders that underwent somatic maturation. However, the time and costs necessary to prepare a library for each new antigen may urge to look for alternatives such as those offered by large one-pot libraries. Here we describe how to obtain a suitable naïve library using material from nonimmunized llamas. Despite the lack of somatic maturation, the selection based on phage display allowed to isolate from such naïve libraries VHHs with affinity in the subnanomolar range and suitable for the standard immunoapplications.

MeSH terms

  • Animals
  • Camelids, New World / genetics*
  • Camelids, New World / immunology*
  • Cell Separation / methods
  • Cell Surface Display Techniques*
  • DNA, Complementary
  • Gene Library*
  • Immunoglobulin Heavy Chains / genetics
  • Immunoglobulin Variable Region / genetics
  • Lymphocytes / cytology
  • Lymphocytes / metabolism
  • Polymerase Chain Reaction
  • RNA / isolation & purification
  • Single-Domain Antibodies / genetics*
  • Single-Domain Antibodies / immunology*
  • Transformation, Bacterial

Substances

  • DNA, Complementary
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Variable Region
  • Single-Domain Antibodies
  • RNA