title: Strain-specific monoclonal antibodies to the E2 protein of classical swine fever virus, Paderborn strain.
abstract: Monoclonal antibodies (MAbs) against the E2 protein of classical swine fever virus (CSFV) are useful for diagnosis and strain characterization. A purified, baculovirus-expressed CSFV E2 protein from the Paderborn strain was formulated with a saponin adjuvant and successfully used to induce an antigen-specific immune response in mice. After cell fusion a panel, designated F92G, of 12 mouse hybridomas (5-2, 11-1, 14-1, 25-2, 28-2, 31-1, 34-1, 35-2, 37-3, 38-2, 39-1, 41-1) producing CSFV-E2 specific MAbs were selected based on their Ig subclass and secretion level (μg IgG/mL). Nine IgG 1/k, two IgG 2b/k, and one IgG 2a/k MAbs were further characterized using immunoperoxidase reactivity, ELISA, and Western blot analysis. Immunoglobulin concentration-dependent immunoperoxidase and ELISA reactivity was observed for some of the MAbs with certain antigens. In general there were several reactivity patterns exhibited by the MAbs, with CSFV strains representing different genetic subgroups (by immunoperoxidase staining) and recombinant antigens (by ELISA). It was interesting to note that in some cases the strain-specific reactivity of a MAb was dependent on the test, thereby providing a clue regarding the nature of the binding site.
authors.affiliation: National Centre for Animal Disease, Canadian Food Inspection Agency, Winnipeg, Manitoba, Canada.
authors.forename: Mariko
authors.initials: M
authors.lastname: Moniwa
authors.forename: Lizhong
authors.initials: L
authors.lastname: Luo
authors.forename: Kevin
authors.initials: K
authors.lastname: Hills
authors.forename: Krista
authors.initials: K
authors.lastname: Nishi
authors.forename: Erin
authors.initials: E
authors.lastname: Macleod
authors.forename: John
authors.initials: J
authors.lastname: Pasick
authors.forename: Marta
authors.initials: M
authors.lastname: Sabara
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000276:immunology*; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D016367:Baculoviridae / Q000235:genetics; D001666:Binding Sites, Antibody; D015153:Blotting, Western; D006692:Classical Swine Fever Virus / Q000737:chemistry / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors; D006825:Hybridomas / Q000276:immunology; D007114:Immunization; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000276:immunology*; D051379:Mice; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D013045:Species Specificity; D013552:Swine; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
chemical_list: D000276:Adjuvants, Immunologic; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C087086:glycoprotein E2, classical swine fever virus