Production and characterization of monoclonal antibodies against bovine leukaemia virus using various crude antigen preparations: A comparative study

A total of 59 monoclonal antibodies (mAbs) specific against the bovine leuk aemia virus (BLV) using different antigen preparations was produced. The fi ve antigen preparations for immunizing mice were: live cells (CEL), sonicat ed and ultracentrifuged cells (SOC), cell lysates (LYS), semi-purified BLV (PV), and formalin-treated cells (FOR) from two cell lines permanently infe cted with BLV (FLK-BLV and BLV-bat(2)). These viral component presentations were selected to obtain mAbs against specific BLV proteins: located on the cell surface (FOR and GEL), in free virus particles (PV) and intracellular viral proteins (SOC and LYS). Two antigen preparations (SOC and LYS) were lethal to the mice following the intravenous and intrasplenic routes. Six f usions were performed in this study that rendered specific antibodies again st BLV. The highest number of hybridomas was produced with SOC; however, th e majority of the hybridomas produced (> 90 %) were against cellular protei ns. Even though immunization with PV gave the lowest number of hybridomas, the majority of them were specific against BLV. Based on the reactivity of the mAbs in Western blot (WB), we classified the mAbs into five groups, nam ely anti (39 mAbs), anti-gp30TM (six mAbs), anti-Pr72(env) (nine mAbs), and -Pr66(gag-pro) (one mAb) and anti-Pr-gag (four mAbs). A very high percentag e of the mAbs produced (48 of 59) reacted with gp51SU, suggesting that this is the most immunogenic and accessible BLV protein presented in the differ ent antigen preparations. The majority of our mAbs recognized more than one band in WB, suggesting that, aside from reacting with mature proteins, the mAbs also recognized viral precursors.