Preparation of Human Salivary σ-Amylase Specific Monoclonal Antibody

Abstract

A mouse hybridoma cell line which produced an anti-human salivary α-amylase monoclonal antibody was obtained by fusion between mouse spleen cells immunized with human salivary α-amylase and mouse myeloma cells, followed by screening the hybridoma cells by enzyme-linked immunosorbent assay. The hybridoma cell line (27–4–1) secreted IgG. The monoclonal antibody produced by the hybridoma showed no inhibitory effect on the activity of human salivary α-amylase. The specificity and reactivity of this monoclonal antibody were examined by determining the activities of human salivary and pancreatic α-amylases bound to the monoclonal antibody immobilized on polystyrene balls or by enzyme immunoassay with the monoclonal antibody conjugated with β-D-galactosidase. The results revealed that the monoclonal antibody produced by the hybridoma cell line was specific for salivary α-amylase and absolutely unreactive to pancreatic α-amylase.