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

使用 ras 基因产物特异性单克隆抗体开发癌症诊断

• 批准号: 60870082
• 负责人: OHTSUKA Eiko
• 金额: $ 3.84万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research
• 财政年份: 1985
• 资助国家: 日本
• 起止时间: 1985 至 1986
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-60870082/
• 关键词: c-Ha-ras gene; Synthetic gene; Monoclonal antibodies; GDP binding activity; GTPase活性; P21; GDP結合蛋白質

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.

本研究旨在制备针对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，我们合成了编码12-Gly、12-Val或61-Lau的基因片段，编码正常和活化的c-Ha-ras蛋白。将合成的基因连接到Trp启动子下游的表达载体中，并在大肠杆菌中表达。用离子交换层析和凝胶过滤的方法对表达的蛋白进行了定量分离。这些蛋白质被用来制造单抗，方法是注射到小鼠体内，并将小鼠的脾细胞与骨髓瘤细胞杂交。对抗体产生细胞进行筛选和克隆。从识别c-Ha-ras(Val-12)和c-Ha-ras(Gly-12)的杂交瘤中获得抗c-Ha-ras(Val-12)。这种抗体将被用来开发一种检测过度产生c-Ha-ras蛋白的癌症的方法。

项目成果

K. Miura, Y. Inoue, H. Nakamori, S. Iwai, E. Ohtsuka, M. Ikehara, S. Noguchi, and S. Nishimura: "Synthesis and Expression of a Synthetic Gene for the Activated c-Ha-ras Protein" Jpn. J. Cancer Res. (Gann). 77. 45-51 (1986)

K. Miura、Y. Inoue、H. Nakamori、S. Iwai、E. Ohtsuka、M. Ikehara、S. Noguchi 和 S. Nishimura：“激活 c-Ha-ras 蛋白的合成基因的合成和表达

Jpn.J.Cancer Res.77-1. (1986)

Jpn.J.Cancer Res.77-1。

K.Miura;Y.Inoue;H.Nakamori;S.Iwai;E.Ohtsuka;M.Ikehara;S.Noguchi;S.Nishimura: Jpn.J.Cancer Res.(Gann). 77. 45-51 (1986)

K.Miura；Y.Inoue；H.Nakamori；S.Iwai；E.Ohtsuka；M.Ikehara；S.Noguchi；S.Nishimura：Jpn.J.Cancer Res.（江恩）。