Differentiation of two bovine lentiviruses by a monoclonal antibody on the basis of epitope specificity.
Bovine immunodeficiency virus (BIV) and Jembrana disease virus (JDV) are bovine lentiviruses that are closely related genetically. A recombinant fusion protein containing the capsid protein of BIV expressed in Escherichia coli was used to immunize mice and produce monoclonal antibodies. Six hybridomas specific for BIV capsid protein were identified, and one antibody, designated 10H1, was characterized further. Competitive binding assays were performed to analyze the topography of antigenic determinants by enzyme-linked immunosorbent assay and demonstrated the existence of at least three distinct antigenic determinants on capsid protein. The monoclonal antibody reacted specifically with both BIV capsid and the recombinant fusion protein in Western immunoblot analyses. However, it did not react with the recombinant capsid fusion protein of JDV, indicating that BIV contains at least one unique epitope in the capsid protein that is absent in JDV. Further mapping of the epitope by chemical cleavage analysis identified that the epitope is located at the 6.4-kDa N terminus of the 29-kDa capsid protein. This monoclonal antibody assay will be valuable for distinguishing the two closely related lentiviruses by Western blotting.
Departments of Diagnostic Medicine/Pathobiology, Kansas State University, Manhattan, KS 66506, USA.
L
L
Zheng
S
S
Zhang
C
C
Wood
S
S
Kapil
G E
GE
Wilcox
T A
TA
Loughin
H C
HC
Minocha
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000951:Antigens, Neoplasm; D002213:Capsid / Q000276:immunology; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000276:immunology; D003937:Diagnosis, Differential; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D006825:Hybridomas; D016087:Immunodeficiency Virus, Bovine / Q000276:immunology* / Q000302:isolation & purification; D007145:Immunoglobulin kappa-Chains; D016180:Lentivirus Infections / Q000175:diagnosis* / Q000276:immunology / Q000662:veterinary; D051379:Mice; D008807:Mice, Inbred BALB C; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D000939:Epitopes; D007145:Immunoglobulin kappa-Chains; C046028:myeloma-associated membrane antigen KMA