MONOCLONAL-ANTIBODIES RAISED AGAINST CARP MYOSIN

Several cell lines producing monoclonal antibodies against carp fast m uscle myosin were obtained by fusion of immunized BALB/c mouse spleen cells with syngenetic NS-1 myeloma cells and subsequent cloning by the limiting dilution method. Monoclonal antibodies, which were obtained when myosin was injected to mice as an antigen, recognized myosin subf ragment-1 (S1). Further assignments for antigenic sites in the S1 mole cule revealed that antibodies from 35A11 and 41C2 cell lines were spec ific to 50 and 25 kDa fragments of S1 tryptic digests, respectively. N o cell line producing monoclonal antibody specific to the 20 kDa fragm ent of S1 tryptic digests was established. On the other hand, cell lin es for monoclonal antibodies against myosin rod were established only when the rod was administered to mice as an antigen. The 37G3 cell lin e produced the antibody specific to myosin rod and reacted with its li ght meromyosin part. However, all monoclonal antibodies thus raised fa iled to discriminate between two types of myosin from carp acclimated to 10 and 30-degrees-C, irrespective of myosin S1 or rod.