Using various antigen preparations to produce monoclonal antibodies against bovine leukaemia virus (BLV) gp51SU

The objective of this study was to compare different antigen preparations to produce monoclonal antibodies against bovine leukaemia virus gp51SU. The four antigen preparations for immunization of BALB/c mice were: CL: BLV-FLK cell lysate, UF: a fraction of CL (between 30 and 100 kDa), WVP: whole virus particles and SP: with ion exchange chromatography, gp51SU was semipurified. A total of nine successful fusions were performed which resulted in production of 23 monoclonal antibodies (mAbs) specific against gp51SU. The highest ratio of specific hybridoma colonies in each fusion was with SP preparation. Based on the reactivity of the mAbs in Western blotting, mAbs were classified into four groups: anti-gp51SU (23 mAbs), anti-gp30TM (8 mAbs), anti-Pr72 (5 mAbs) and antibodies against other viral proteins (7 mAbs). Some of the anti-gp51SU mAbs reacted with more than one band in Western blotting, suggesting that these colonies recognized not only gp51SU but also its precursors.