Article

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

Department of Molecular and Cell Biology, Institute of Medical Sciences, University of Aberdeen, Aberdeen AB25 2ZD, Scotland, UK.
Molecular Immunology (impact factor: 2.9). 10/2003; 40(1):25-33.
Source: PubMed

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.

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Keywords

affinity
 
alternative route
 
cartilaginous fish
 
favorable characteristics
 
Ginglymostoma cirratum
 
heavy-chain homodimer
 
heavy-chain immunoglobulins
 
independent domains
 
intact antigen-specific binders
 
light chains
 
minimal antigen-binding fragments
 
model antigen hen egg-white lysozyme
 
nanomolar range
 
novel immunoglobulin isotype novel antigen receptor
 
nurse sharks
 
selected variable domains
 
single-domain library
 
variable domain
 
variable regions
 

Helen Dooley