Production of Monoclonal Antibody Probes Specific for Nonbasement Membrane Glomerular Capillary Wall Antigens in the Rat

To examine the characteristics of discontinuously represented antigens within the rat glomerular capillary wall, we have developed monoclonal antibodies against basement membrane-depleted glomerular plasma membrane preparations. These reagents exhibited granular binding patterns on normal rat glomeruli when examined by indirect immunofluorescence. By limited purification of the primary immunogen and the use of this material for the initial screening by micro-ELISA, the production and selection of monoclonal antibodies (McAbs) of the desired specificity was enhanced. Intrasplenic immunisation was usefully employed to reduce the time interval from immunisation to fusion without lessening the percentage of positive hybridomas generated. The potentiality for in situ immune responses to discretely arrayed non-basement membrane glomerular capillary wall antigens which produce granular immunofluorescence is further exemplified by these studies.