title: Expression, purification and antigenicity of Neospora caninum-antigens using silkworm larvae targeting for subunit vaccines.
abstract: Infection of Neospora caninum causes abortion in cattle, which has a serious worldwide impact on the economic performance of the dairy and beef industries. Now, inexpensive and efficacious vaccines are required to protect cattle from neosporosis in livestock industry. In this study, N. caninum surface antigen 1 (SAG1) and SAG1-related sequence 2 (SRS2) were expressed in hemolymph of silkworm larvae as a soluble form. Expressed SAG1 and SRS2 clearly showed antigenicity against N. caninum-positive sera of cow. SAG1 and SRS2 were purified to near homogeneity from hemolymph of silkworm larvae using anti-FLAG M2 antibody agarose: approximately 1.7 mg of SAG1 from 10 silkworm larvae and 370 μg of SRS2 from 17 silkworm larvae. Mice that were injected by antigens induced antibodies against SAG1 and SRS2. This study indicates that it is possible that this silkworm expression system leads to a large-scale production of N. caninum-antigens with biological function and low production cost. Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid expression system paves the way to produce largely and rapidly these recombinant antigens for its application to subunit vaccines against neosporosis in cattle.
authors.affiliation: Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, Shizuoka 422-8529, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Otsuki
authors.forename: Jinhua
authors.initials: J
authors.lastname: Dong
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Kato
authors.forename: Enoch Y
authors.initials: EY
authors.lastname: Park
mesh_terms: D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D000954:Antigens, Surface / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D012831:Bombyx / Q000378:metabolism / Q000821:virology*; D002417:Cattle; D003048:Coccidiosis / Q000276:immunology / Q000469:parasitology / Q000662:veterinary*; D015870:Gene Expression; D006458:Hemolymph; D007114:Immunization; D007814:Larva; D051379:Mice; D008807:Mice, Inbred BALB C; D018688:Neospora / Q000235:genetics / Q000276:immunology*; D017924:Nucleopolyhedroviruses / Q000235:genetics; D021382:Protein Sorting Signals; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D016052:Protozoan Vaccines / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D019076:Transgenes; D022223:Vaccines, Subunit / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D000953:Antigens, Protozoan; D000954:Antigens, Surface; D021382:Protein Sorting Signals; D015800:Protozoan Proteins; D016052:Protozoan Vaccines; D011993:Recombinant Fusion Proteins; C115390:SAG1 protein, Neospora caninum; C115389:SRS2 protein, Neospora caninum; D022223:Vaccines, Subunit