神经元特异性烯醇化酶的快速检测方法研究

拟以双抗体夹心法制备快速检测试条，检测脑中风疑似患者体液中的NSE水平，从而达到快速诊断目的。本研究通过DEAE-Sephadex A-50柱层析法完成了牛脑NSE的提取纯化，并建立了具有抗ANSE活性的Anti-NSE MoAb AA10和AB3单克隆抗体细胞株，完成了单克隆抗体的筛选、制备、工作单抗配对等方面的工作。

Study of a new method for detecting NSE in body fluid

Neuron-specific enolase (NSE) represents the gamma-isoforms of the dimeric glycolytic enzyme enolase. NSE is predominantly found in neurons and neuroendocrine cells and has proven to be a marker for monitoring brain damage. The information obtained according to the method can be provided at the critically important early stages of a stroke, e.g., within the first three to six hours after the onset of symptoms since the analysis of the patient's body fluid can be carried out in several minutes after the body fluid is collected. By measuring the marker in samples of body fluid taken at different points in time the progress of the stroke can be ascertained. Reported in this study is the process of the purification of NSE from bovine brain. And the monoclonal antibody against gamma-enolase were raised in mice and selected for optimal performance. The antibody combination of choice was MoAb AA_(10) for capturing and MoAb AB_3 for detection which is labeled by golden colloid. The resulting assay is a one-step strip test of the sandwich type.