Preparation of monoclonal antibodies to chicken bone phosphoproteins

Monoclonal and polyclonal antibodies were raised against 150k, 60k, 32k, and 15k phosphoprotein components of 14-week chicken bone. Hybridomas were prepared by immunizing Balb/c mice with 150k, 60k, or 32k phosphoproteins followed by fusion of their spleen cells with X63-Ag8.653 myeloma cells. Polyclonal antibodies to the 60k, 32k, and 15k phosphoprotein components were produced in immunized rabbits. Immunological cross-reactivity between antigen and antibody was examined by enzyme-linked immunosorbent assay (ELISA) and dot-blotting. There was cross-reactivity between the 150k, 32k, and 15k phosphoprotein components and between the 150k, 90k, and 60k components. The antibodies raised against 60k component do not bind 32k and 15k antigens.