Application of cellular ELISA (CELISA) to the detection of human monoclonal autoantibodies.

ABSTRACT

This work describes the application of cellular enzyme-linked immunosorbent assay (CELISA) for the detection of both polyreactive and monospecific human monoclonal antibodies against autoantigens. The CELISA is ideally suited for the screening of a large number of hybridoma culture supernatants, being, in this way, superior to other methods commonly used for the detection of autoantibody activity, such as indirect immunofluorescence on tissue sections and slide cell preparations, in terms of speed and sensitivity. This assay demonstrated higher sensitivity than ELISA using autoantigenic extracts from rabbit thymus, human spleen, nucleoprotamine, and salmon sperm nuclei, and enzyme immunoassays using ssDNA, dsDNA, and affinity purified autoantigens as substrate. The CELISA has been also successfully applied to the detection of autolymphocytotoxic antibody activity in heterohybridoma supernatants.