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抗Ras GTP酶激活蛋白的非中和性单克隆抗体:制备、特性鉴定及其在酶免疫分析中的应用

Non-neutralizing monoclonal antibodies against Ras GTPase-activating protein: production, characterization and use in an enzyme immunometric assay.

作者信息

Mollat P, Zhang G Y, Frobert Y, Zhang Y H, Fournier A, Grassi J, Thang M N

机构信息

INSERM U. 245, Hôpital Saint-Antoine, Paris, France.

出版信息

Biotechnology (N Y). 1992 Oct;10(10):1151-6. doi: 10.1038/nbt1092-1151.

Abstract

We studied several monoclonal antibodies (mAbs) raised against the 100 kD Ras GTPase activating protein (p100-GAP), which was purified from human placenta. These antibodies recognized p120-GAP and p100-GAP in native and in denatured forms. The most reactive, GP15 and GP200, both recognized distinct epitopes and did not neutralize GTPase stimulatory activity. These two mAbs were selected for a two-site enzyme immunoassay, using covalent conjugates of the antibodies coupled to the tetrameric form of acetylcholinesterase as tracer. This assay was used to quantify Ras-GAP in both normal and tumor tissues and cell extracts.

摘要

我们研究了几种针对从人胎盘中纯化得到的100 kD Ras GTP酶激活蛋白(p100-GAP)产生的单克隆抗体(mAb)。这些抗体可识别天然和变性形式的p120-GAP和p100-GAP。反应性最强的GP15和GP200都识别不同的表位,且不中和GTP酶刺激活性。选择这两种单克隆抗体用于两点酶免疫测定,使用与乙酰胆碱酯酶四聚体形式偶联的抗体共价缀合物作为示踪剂。该测定法用于定量正常组织、肿瘤组织和细胞提取物中的Ras-GAP。

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