Wouter van Wyngaardt1 , Teresiah Malatji1, 2 , Cordelia Mashau1 , Jeanni Fehrsen1 , Frances Jordaan1 , Dubravka Miltiadou1 and Dion H du Plessis1
1 Immunology Division, Onderstepoort Veterinary Institute, Onderstepoort, 0110, Republic of South Africa
2 Present address: John and Kernick, PO Box 3511, Midrand, 1685, Republic of South Africa BMC Biotechnology 2004, 4:6 doi:10.1186/1472-6750-4-6
The electronic version of this article is the complete one and can be found online at: http://www.biomedcentral.com/1472-6750/4/6 © 1995-2006 van Wyngaardt et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.
Abstract
Antibody fragments selected from large combinatorial libraries have numerous applications in diagnosis and therapy. Most existing antibody repertoires are derived from human immunoglobulin genes. Genes from other species can, however, also be used. Because of the way in which gene conversion introduces diversity, the naïve antibody repertoire of the chicken can easily be accessed using only two sets of primers.
With in vitro diagnostic applications in mind, we have constructed a large library of recombinant filamentous bacteriophages displaying single chain antibody fragments derived from combinatorial pairings of chicken variable heavy and light chains. Synthetically randomised complementarity determining regions are included in some of the heavy chains. Single chain antibody fragments that recognise haptens, proteins and virus particles were selected from this repertoire. Affinities of three different antibody fragments were determined using surface plasmon resonance. Two were in the low nanomolar and one in the subnanomolar range. To illustrate the practical value of antibodies from the library, phage displayed single chain fragments were incorporated into ELISAs aimed at detecting African horsesickness and bluetongue virus particles. Virus antibodies were detected in a competitive ELISA.
The chicken-derived phage library described here is expected to be a versatile source of recombinant antibody fragments directed against a wide variety of antigens. It has the potential to provide monoclonal reagents with applications in research and diagnostics. For in vitro applications, naïve phage libraries based on avian donors may prove to be useful adjuncts to the selectable antibody repertoires that already exist. | 
