Production and characterization of monoclonal antibody to bovine leukemia virus gp51-SU

Abstract

Enzootic bovine leukemia (EBL) is a disease of cattle with worldwide distribution. The bovine leukemia virus (BLV), is a typical type C retrovirus, has four genes: gag, pol, pro and env, flanked by long terminal repeats (LTR) and lacking oncogenes. Gene env encodes a precursor protein named Pr72 env which undergoes glycosylation and lysis, giving rise to an envelope glycoprotein gp51-SU with a total of 12 epitopes. The objective of the present study was to produce a panel of monoclonal antibodies (mAbs) against BLV-gp51-SU following the standard immunization protocols. The antigen preparations included cellular lysates, a fraction of cellular lysates to study antibodies against BLV proteins precursors, whole virus particles and semipurified gp51 -SU. Both route of injection (SC and IP) resulted in hyperimmunized mice. As indicated by indirect-ELISA and also yielded high fusion efficiency. The hybridoma colonies were stable and had a tendency to save their ability to secrete antibodies. Fusions produced five positive colonies in screening tests, two of which were against gp51-SU. The targeted hydrophobic domain of the gp51-SU molecule was detected by one of the colonies of mAbs. The two interested mAbs were used for immunohistochemistry on lymph node sections of a cow with lymphosarcoma which can detect immunoreactive materials in sections and also in H&E staining. We conclude the produced panel of mAbs is able to be used in detection of gp51-SU and a new epitope identified on this molecule could be used in IHC method for diagnosis of BLV infection. © IDOSI Publications, 2011.