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

本研究旨在制备抗c-Ha-ras基因产物的抗体。这些抗体可用于癌症的诊断。为了制备c-Ha-ras蛋白，合成编码正常和活化c-Ha-ras蛋白的基因，编码12-gly、12-瓦尔或61-lau的基因片段。将合成基因连接到表达载体中trp启动子的下游，并在大肠杆菌中表达。通过离子交换层析和凝胶过滤等方法对表达的蛋白质进行定量分离。这些蛋白质通过注射到小鼠体内并使其脾细胞与骨髓瘤细胞杂交来生产单克隆抗体。筛选并克隆抗体产生细胞。从杂交瘤中获得的抗c-Ha-ras（瓦尔-12）识别c-Ha-ras（瓦尔-12）以及c-Ha-ras（gly-12）。该抗体将用于开发一种检测过度产生c-Ha-ras蛋白的癌症的方法。

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.