The Development of Chicken Recombinant Single-chain Fv (ScFv) Antibody Reactive with Sporozoite Antigen of Eimeria spp. which Causes Avian Coccidiosis

Abstract

The chicken monoclonal antibody (mAb), 13C8, reacts with sporozoite antigens of Eimeria spp. which causes avian coccidiosis. Since this mAb was produced at low amount due to genetic instability of chicken hybridoma, a recombinant 13C8 single-chain Fv (ScFv) antibody was constructed by amplification of the variable domain of heavy (VH) and light chain (VL) genes of antibody derived from chicken hybridoma. The constructed 13C8 ScFv was successfully expressed in E. coli and purified as a soluble form. In ELISA analysis, this recombinant 13C8 ScFv antibody showed antigen binding activity as the original mAb. In addition, nucleotide sequence comparison of 13C8 gene to the germline chicken VL and VH genes suggested that the gene conversion with Vλ and VH pseudogenes might contribute to the diversification of VL and VH genes in chickens.(