Production and evaluation of the specific nature of monoclonal antibodies against bovine leukaemia virus

The aim of this study was to prepare monoclonal antibodies (mabs) directed against major immuno-dominant epitope of BLV antigens. Myeloma cells Sp2/0 and spleen cells of BLV immunised BALB/c mice were used to prepare hybridom a cell clones by the usual hybridoma technology. Out of the 13 resultant cl ones, the supernatants of 11 showed a strong secretory activity in the ELIS A assay, with intact BLV virus as the antigen. After the re-cloning procedu re, 22 new clones were selected by the use of the limited cloning method. S upernatants from these clones were examined by ELISA for their secretory ac tivity. Nineteen positive supernatants were collected and then examined for the specificity of mabs. Three methods were employed to achieve this: west ern blot with saccharose purified BLV antigens, immunoperoxidase mono-layer assay (IPMA) with FLK cells infected with BLV and immuno-peroxidase infect ivity assay (IPIA) with CC81 indicatory cells, co-cultivated with FLK cells , The mabs from the examined supernatants reacted with gp51 protein and mer e classified as being IgG(1) isotype. Two clones, specific for gp51 by the western blot, produced distinct colour reactions on micro-plates coated wit h FLK or CC81/FLK cells. The resulted mabs will be useful for further devel oping diagnostic methods for BLV infection.