Discrimination of determinant specificity of two encephalomyocarditis virus neutralizing monoclonal antibodies by competition, mixed neutralization and anti-idiotypic antibodies

Abstract

To investigate the epitope(s) on encephalomyocarditis virus (EMCV) involved in neutralization, two neutralizing monoclonal antibodies (MAs) (MA UM 21.1 and MA UM 21.2) were tested in a competition binding assay (CBA), a mixed neutralization test and an enzyme immunoassay (EIA) with specificity for the detection of idiotypes on MAs. With a CBA in cell culture, using EMCV infected L cell monolayers as binding antigen, strong homologous competition was observed between unlabelled MAs and horse radish peroxidase (HRPO-) labelled MAs but considerable heterologous competition did also occur, especially between the unlabelled MA UM 21.1 and HRPO-labelled MA UM 21.2. In the mixed neutralization test (50% plaque reduction) preincubation with slightly neutralizing or non-neutralizing doses of MA UM 21.2 had no diminishing effect on the neutralizing capacity of MA UM 21.1, the PRT 50 value remains in all cases −10log titre of 5.8. Furthermore rabbit polyclonal antibodies against the idiotypes of MAs UM 21.1 and UM 21.2 did not cross react in the EIA. In conclusion both MAs recognize different viral determinants as indicated by the results obtained with a CBA, a mixed neutralization test and an EIA for detection of idiotypes on MAs.