A new approach to monoclonal antibody production. In vitro immunization with antigens on nitrocellulose using Drosophila myosin heavy chain as an example.

A methodology of broad applicability is described for the production of monoclonal antibodies to antigens with interesting developmental properties using Drosophila myosin heavy chain to exemplify the procedure. The technique consists of two parts. (1) Identification of the antigen with any tool which allows its detection on a Western blot. In the present case, this was achieved by monospecific polyclonal antibodies, prepared as described by Smith and Fisher (J. Cell Biol. (1984) 99, 20), which usefully define the characteristic properties of the antigen. (2) In vitro immunization of splenocytes with antigen on nitrocellulose whose position on Western blots was detected using monospecific polyclonal antibodies, followed by generation of monoclonal antibodies. A total of 19 hybridomas were selected by immunohistochemistry screening and at least six of them were directed against antigens possessing the specific characteristic properties of myosin heavy chain.