Monoclonal anti‐V H antibodies recognize a common V H determinant expressed on immunoglobulin heavy chains from various species

Abstract Our previous work using rabbit antibodies to the variable region of MOPC315 myeloma heavy chain (V H ) has indicated the existence of framework determinant(s) common to many murine heavy chains. Here we report the characterization of anti‐V H monoclonal antibodies (mAb) prepared in an attempt to elucidate the nature of the common V H determinant. We immunized AKR/J mice with a purified V H 315 fragment and generated somatic cell hybrids by the fusion of the immune AKR/J splenocytes with the NS1 myeloma cells. Thirty‐seven common anti‐V H and 57 subgroup V H I‐specific hybridomas have been established and characterized. Whereas the anti‐subgroup mAb seemed to react with a determinant unique to the MOPC315 (mouse V H I) subgroup, all the anti‐V H mAb reacted with myeloma heavy chains of different V H subgroups, class and allotypes. Antibody competition studies revealed that the V H subgroup determinants are distinct from the common V H determinants and that both were also recognized by the rabbit polyclonal antibodies. The common V H determinants were found to be “hidden” determinants on intact immunoglobulin molecules being exposed only on isolated heavy chains. Furthermore, they are sequential determinants since they are preserved on fully denatured heavy chains. The common V H determinants are shared by immunoglobulins of a wide range of vertebrates from amphibia to man and thus represent antigenic structures which were highly conserved throughout evolution.