MONOCLONAL-ANTIBODIES OF PREDEFINED SPECIFICITY DETECT ACTIVATED RAS GENE-EXPRESSION IN HUMAN MAMMARY AND COLON CARCINOMAS
MONOCLONAL-ANTIBODIES OF PREDEFINED SPECIFICITY DETECT ACTIVATED RAS GENE-EXPRESSION IN HUMAN MAMMARY AND COLON CARCINOMAS
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DOI：
10.1073/pnas.81.16.5227
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发表时间：
1984-01-01
期刊：
影响因子：
--
通讯作者：
SCHLOM, J
中科院分区：
文献类型：
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作者：
HAND, PH;WUNDERLICH, ATD;SCHLOM, J
通过利用反映Hu-rasT 24基因产物的氨基酸位置10-17的合成肽作为免疫原来产生预定特异性的单克隆抗体（MAb）。这些单克隆抗体，命名为RAP-1至RAP-5（RA，ras; P，肽），显示与ras基因产物p21反应。由于预测Hu-ras反应性决定簇（位置10-17）在p21分子的三级结构内，因此用福尔马林或戊二醛变性细胞提取物或组织切片增强RAP MAb的结合并不意外。当石蜡包埋福尔马林固定的组织切片和抗生物素蛋白-生物素复合物免疫过氧化物酶法被使用; RAP单克隆抗体明确定义增强的ras p21在大多数人结肠癌和乳腺癌的表达。纤维腺瘤和纤维囊性疾病患者的大多数异常导管和小叶是阴性的，所有正常的乳腺和结肠上皮检查。本文报道的结果构成了ras翻译产物定量放射免疫分析的基础，并提供了一种方法来评价正常组织和良性、癌前和恶性病变的单个细胞内ras p21的表达。
Monoclonal antibodies (MAb) of predefined specificity were generated by utilizing a synthetic peptide reflecting amino acid positions 10-17 of the Hu-rasT24 gene product as immunogen. These MAb, designated RAP-1 through RAP-5 (RA, ras; P, peptide), were shown to react with the ras gene product p21. Since the Hu-ras reactive determinants (positions 10-17) were predicted to be within the tertiary structure of the p21 molecule, it was not unexpected that denaturation of cell extracts or tissue sections with Formalin or glutaraldehyde enhanced binding of the RAP MAb. When paraffin-embedded Formalin-fixed tissue sections and the avidin-biotin complex immunoperoxidase method were used; the RAP MAb clearly defined enhanced ras p21 expression in the majority of human colon and mammary carcinomas. The majority of all abnormal ducts and lobules from fibroadenoma and fibrocystic disease patients were negative, as were all normal mammary and colonic epithelia examined. The findings reported here form the basis for quantitative radioimmmunoassays for a ras translational product and provide a means to evaluate ras p21 expression within individual cells of normal tissues and benign, premalignant, and malignant lesions.