Sias P E, Kotts C E, Vetterlein D, Shepard M, Wong W L
Department of Immunology Research and Assay Technologies, Genentech Inc., So. San Francisco, CA 94080.
J Immunol Methods. 1990 Aug 28;132(1):73-80. doi: 10.1016/0022-1759(90)90400-p.
The HER2/neu proto-oncogene encodes a receptor that belong to the tyrosine-specific protein kinase family. Amplification of the HER2 gene in patients with breast and ovarian cancer has been shown to predict poorer survival rates. In order to understand the role of HER2 in malignant and normal cells, it is necessary to devise assays that can quantitate expression levels of the HER2 gene product (p185HER2) in production samples, biopsy specimens and biological fluids. We have developed a simple, quantitative ELISA that uses two monoclonal antibodies directed against the extracellular domain of the HER2 gene product, p185HER2 (HER2 ECD). The assay has a detection range of 0.25-120 ng/ml, is precise and sensitive. The ability of this assay to detect biologically active rHER2 ECD is demonstrated by its correlation to a growth inhibitory bioassay (r = 0.92). The sandwich ELISA can also accurately quantitate rHER2 ECD in mouse and monkey serum. This assay should be useful for quantitating low levels of circulating rHER2 ECD in animals in which rHER2 ECD is being used as antigen for immunotherapy and in patients which 'shed' receptor.
HER2/neu原癌基因编码一种属于酪氨酸特异性蛋白激酶家族的受体。乳腺癌和卵巢癌患者中HER2基因的扩增已被证明预示着较差的生存率。为了了解HER2在恶性细胞和正常细胞中的作用，有必要设计出能够对生产样品、活检标本和生物体液中HER2基因产物（p185HER2）的表达水平进行定量的检测方法。我们开发了一种简单的定量酶联免疫吸附测定（ELISA），它使用两种针对HER2基因产物p185HER2（HER2胞外域）胞外域的单克隆抗体。该检测方法的检测范围为0.25 - 120 ng/ml，精确且灵敏。该检测方法检测生物活性重组人HER2胞外域（rHER2 ECD）的能力通过其与生长抑制生物测定的相关性得到证明（r = 0.92）。夹心ELISA还能准确地对小鼠和猴血清中的rHER2 ECD进行定量。该检测方法对于定量在将rHER2 ECD用作免疫治疗抗原的动物以及“脱落”受体的患者中循环的低水平rHER2 ECD应该是有用的。
