Abstract
More than two hundred and fifty-five monoclonal antibody (MoAb)-secreting hybridomas against 3 plant luteoviruses, 2 plant reoviruses and a potyvirus were produced. The hybridomas for potato leafroll virus, beet western yellows virus, tobacco necrotic dwarf virus, rice dwarf virus, rice ragged stunt virus and potato virus Y-ordinary strain, were screened by four different procedures of enzyme-linked immunosorbent assay (ELISA); procedure 1, antigen adsorption indirect ELISA (AAI-ELISA) in which purified virus in phosphate buffered saline (PBS) at pH 7.4 was adsorbed onto the microplate wells, procedure 2, AAI-ELISA in which purified virus in sodium carbonate-bicarbonate buffer at pH 9.6 was adsorbed onto the microplate wells, procedure 3, indirect double antibody sandwich ELISA (IDAS-ELISA) in which polyclonal antibody was used as trapping antibody and purified virus preparations diluted in PBS-T (containing Tween-20) as antigens were used, procedure 4, IDAS-ELISA in which polyclonal antibody was used for trapping antibody and crude saps of virus infected plants extracted in PBS-T as antigens were used. Based on the MoAb reactivities against homologous viruses in four different ELISA procedures, MoAb-secreting hybridomas were divided into ten groups. Using purified MoAbs from ascitic fluids, direct double antibody sandwich ELISA (DAS-ELISA) was examined for the detection of virus antigens in infected plants. All the MoAbs reacted in DAS-ELISA belonged to group 1 in which the MoAbs were reactive in each four of the screening procedures 1-4, or group 2 in which MoAbs were reactive in three of screening procedures 1, 3 and 4. These results indicate that MoAbs being reactive in DAS-ELISA can be readily selected in hybridomas in group 1 or 2.
