Production, Characterization, and Use of Monoclonal Antibodies Against gp51 Protein to Diagnose Bovine Leukemia Virus Infection

Abstract

Enzootic bovine leukosis (EBL) is a retroviral infection that causes persistent lymphocytosis and lymphosarcoma in cattle. The economic importance of infection by bovine leukemia virus (BLV) is due to several factors, including losses in exportation, treatment of secondary infection, and reduction in dairy production. To facilitate the development of a national test that is sensitive, simple, and applicable on a large scale, this work aimed to produce and characterize monoclonal antibodies (mAbs) against gp51 protein from BLV for use in an enzyme-linked immunosorbent assay (ELISA) test. Two hundred seventy-four hybridomas were generated, from which 37 were mAbs secretory clones screened by indirect ELISA. The specificity of the mAbs generated against gp51 was verified by Western blot analysis, and the isotypes were characterized for isotyping in IgG1 and IgM. To evaluate the test, 250 sera were tested by agar gel immunodiffusion and mAb-ELISA. The values obtained for the mAb-ELISA test were 95% sensitivity and 90% specificity.

Development and evaluation of an immunochromatographic assay using a gp51 monoclonal antibody for the detection of antibodies against the bovine leukemia virus

“…The selected MAbs were further tested for their ability to interfere with ligand-receptor binding in a blocking ELISA, as previously described [28]. Western blot and isotype analyses were performed as previously described [34] using the hybridoma with superior blocking ability.…”

Characterization and application of recombinant Bovine Leukemia Virus Env protein

“…Gp51 is the most immunogenic subunit of Env and induces massive expression of specific antibodies in infected animals 20 , 21 . The N-terminal region of gp51 (the first 173 aa 22 ) contains putative receptor binding domain (RBD 16 ) together with conformational epitopes namely F, G 23 and H, involved in infectivity and syncytia formation and two neutralization domains 23 – 28 .…”

Enzyme-Linked Immunosorbent Assays Using the Recombinant gp51 and p24 of Bovine Leukemia Virus for Immunodetection of the Disease

“…Structural proteins p24 and gp51 are potent immunogens and the antibody production can be detected in most of the animals infected with VLB. In experimentally infected animals anti-gp51 antibodies seem to be the first to appear with titres consistently higher than anti-p24 antibodies [7] [8] [9]. Other authors reports that high sensitivity tests are able to detect anti-p24 antibodies before the appearance of anti-gp51 antibodies; and the anti-p24 antibodies could be an equal or greater degree than anti-gp51 [3] [10].…”