title: Development of two murine antibodies against Neospora caninum using phage display technology and application on the detection of N. caninum.
abstract: Neosporosis, caused by an intracellular parasite, Neospora caninum, is an infectious disease primarily of cattle and dogs. It occurs worldwide and causes huge damages to dairy farms. In this study, we immunized mice with recombinant surface-associated protein 1 of N. caninum (rNcSAG1) and developed two novel monoclonal antibodies, A10 and H3, against NcSAG1 using phage-display technology. Both clones bound to purified rNcSAG1 and the half maximal inhibitory concentrations of A10 and H3 are 50 and 72 nM of rNcSAG1, respectively. In immunofluorescence assays, both A10 and H3 Fabs bound to N. caninum parasites. Direct detection of N. caninum parasites was developed firstly using an enzyme-linked immunosorbent assay (ELISA) with A10 and H3. Binding of A10 and H3 antibodies to rNcSAG1 was also inhibited by some certain anti-N. caninum antibodies in the neosporosis-positive cattle sera, suggesting they might bind to the same epitopes of NcSAG1 with those anti-N. caninum antibodies of bovine. These antibodies were demonstrated to have a potential for monitoring the N. caninum parasites in a dairy farm, which may lead to protect livestock from parasite-infection.
authors: Laboratory of Biotechnology, Graduate School of Science and Technology, Shizuoka University, Shizuoka, Japan.
authors: Jinhua
authors: J
authors: Dong
authors: Takahiro
authors: T
authors: Otsuki
authors: Tatsuya
authors: T
authors: Kato
authors: Tetsuya
authors: T
authors: Kohsaka
authors: Kazunori
authors: K
authors: Ike
authors: Enoch Y
authors: EY
authors: Park
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000913:Antibodies, Protozoan / Q000276:immunology*; D000953:Antigens, Protozoan / Q000276:immunology; D002417:Cattle; D061505:Cell Surface Display Techniques; D003048:Coccidiosis / Q000175:diagnosis* / Q000276:immunology / Q000662:veterinary; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D018688:Neospora / Q000276:immunology* / Q000302:isolation & purification*; D015800:Protozoan Proteins / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D015800:Protozoan Proteins