Development of cancer diagnosis using monoclonal antibodies specific for ras gene products

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The present invesigation intends a production of antibodies against products of c-Ha-ras genes. These antibodies can be applied to diagonosis of cancers. In order to prepare c-Ha-ras proteins, genes coding for normal and activated c-Ha-ras proteins were synthesized gene fragments coding for either 12-gly, 12-val or 61-lau. The synthetic genes were joined to downstream of the trp promoter in an expression vector and expressed in E.coli. The expressed proteins were isolated by ion-exchange chromatography and gel filtration in quantity. These proteins were used to produce monoclonal antibodies by injecting to mice and hybridization of their spleen cells with myeloma cells. Antibody producing cells were screened and cloned. Anti c-Ha-ras (val-12) obtained from a hybridoma recognized c-Ha-ras (val-12) as well as c-Ha-ras (gly-12). This antibody are to be used to develop a method to detect cancers overproducing c-Ha-ras proteins.