Submitted to: Avian Immunology Research Group Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 2/7/2001
Publication Date: N/A
Technical Abstract: Chicken hybridoma 6D12 secretes monoclonal antibody (mAb) which identifies an apical complex protein present on the invasive stages of Eimeria. The target antigen identified by the mAb 6D12 is highly conserved among other apicomplexans including Neospora and Cryptosporidia. Culture supernatant from 6D12 hybridoma inhibits the invasion of sporozoites into CD8+ T lymphocytes in vitro indicating that this mAb recognizes a potential receptor protein of Eimeria. In order to express high concentration of recombinant 6D12 ScFv, a single chain fragment variable region (ScFv) was produced by constructing a ScFv gene containing the variable heavy (VH) and light (VL) chain genes. ScFv genes were amplified by PCR using the two primer pairs and the ScFv was assembled by combining the amplified PCR products of VH and VL domains with a flexible linker in a PCR. Recombinant 6D12 ScFv which was expressed in E. coli stained the conoid antigen in an indirect immunofluorescence assay and bound an Eimeria sporozoite antigen in ELISA. These results demonstrate that recombinant 6D12 ScFv recognizes the native conoid antigen on Eimeria.