Quantitation of bovine immunoglobulin isotypes and allotypes using monoclonal antibodies.
A panel of 10 monoclonal antibodies specific for bovine immunoglobulins M, A, G1, G2 and light chains were produced and enzyme-linked immunosorbent assays developed to measure Ig levels in body fluids and culture supernatants using this panel of MAbs. An inhibition ELISA was accurate and sensitive for MAbs of high affinity, detecting levels as low as 10 ng ml-1 of IgM using a high-affinity MAb, IL-A50 (dissociation constant = 1.3 X 10(-11) M). For MAbs of lower affinity (KD of less than 0.25 X 10(-9) M) a sandwich ELISA was more sensitive, detecting 0.1-1.0 microgram ml-1 Ig, provided a conjugate of an anti-light chain MAb was used. Using these ELISA techniques, four pairs of MAbs specific for bovine IgM, IgA, IgG1 and IgG2 respectively, were screened on sera from over 100 cattle of different breeds to determine whether any detected a polymorphic epitope. MAbs IL-A30, IL-A60, IL-A66, IL-A71, IL-A72, IL-A73 and IL-A74 were shown to recognise monomorphic determinants on their respective heavy chains. In contrast, the epitope recognised on the mu-heavy chain by MAb IL-A50, which had previously been shown to be polymorphic, was found to be allelic and inherited under the control of a single gene, probably Cu.
International Laboratory for Research on Animal Diseases, Nairobi, Kenya.
D J
DJ
Williams
J
J
Newson
J
J
Naessens
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002417:Cattle / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary; D000939:Epitopes / Q000276:immunology; D007070:Immunoglobulin A / Q000276:immunology; D007126:Immunoglobulin Allotypes / Q000032:analysis*; D007074:Immunoglobulin G / Q000276:immunology; D007132:Immunoglobulin Isotypes / Q000032:analysis*; D007075:Immunoglobulin M / Q000276:immunology; D007163:Immunosorbent Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D012015:Reference Standards
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007070:Immunoglobulin A; D007126:Immunoglobulin Allotypes; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes; D007075:Immunoglobulin M