Selection and characterization of naturally occurring single-domain (IgNAR) antibody fragments from immunized sharks by phage display

H. Dooley, M. F. Flajnik, Andrew Justin Radcliffe Porter

Research output: Contribution to journalArticle

113 Citations (Scopus)

Abstract

The novel immunoglobulin isotype novel antigen receptor (IgNAR) is found in cartilaginous fish and is composed of a heavy-chain homodimer that does not associate with light chains. The variable regions of IgNAR function as independent domains similar to those found in the heavy-chain immunoglobulins of Camelids. Here, we describe the successful cloning and generation of a phage-displayed, single-domain library based upon the variable domain of IgNAR. Selection of such a library generated from nurse sharks (Ginglymostoma cirratum) immunized with the model antigen hen egg-white lysozyme (HEL) enabled the successful isolation of intact antigen-specific binders matured in vivo. The selected variable domains were shown to be functionally expressed in Escherichia coli, extremely stable, and bind to antigen specifically with an affinity in the nanomolar range. This approach can therefore be considered as an alternative route for the isolation of minimal antigen-binding fragments with favorable characteristics. (C) 2003 Elsevier Science Ltd. All rights reserved.

Original languageEnglish
Pages (from-to)25-33
Number of pages8
JournalMolecular Immunology
Volume40
Issue number1
DOIs
Publication statusPublished - Sep 2003

Keywords

  • single-domain antibody fragment
  • phage display
  • shark
  • novel antigen receptor
  • HEAVY-CHAIN ANTIBODIES
  • RECEPTOR GENE NAR
  • ANTIGEN RECEPTOR
  • SOMATIC HYPERMUTATION
  • LIGHT-CHAINS
  • NURSE SHARK
  • STRUCTURAL-ANALYSIS
  • ESCHERICHIA-COLI
  • REPERTOIRE
  • LYSOZYME

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