IMPEDANCE BASED SENSING OF THE SPECIFIC BINDING REACTION BETWEEN STAPHYLOCOCCUS ENTEROTOXIN-B AND ITS ANTIBODY ON AN ULTRA-THIN PLATINUM FILM
IMPEDANCE BASED SENSING OF THE SPECIFIC BINDING REACTION BETWEEN STAPHYLOCOCCUS ENTEROTOXIN-B AND ITS ANTIBODY ON AN ULTRA-THIN PLATINUM FILM
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DOI：
10.1016/0956-5663(95)96958-2
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发表时间：
1995-09-01
影响因子：
12.6
通讯作者：
STETTER, JR
中科院分区：
文献类型：
--
作者：
DESILVA, MS;ZHANG, Y;STETTER, JR
测量特异性抗原抗体结合反应的免疫生物传感技术对于生物传感器在生物技术、体外诊断、医学和食品技术中的应用发展具有重要意义。构建了免疫生物传感器来测量葡萄球菌肠毒素B（SEE）和抗SEE抗体之间的特异性结合反应。该生物传感器包含共价固定在超薄铂 (Pt) 薄膜上的抗 SEE 生物活性层，超薄铂 (Pt) 薄膜溅射到硅芯片上 100 nm 厚的二氧化硅层上。 Pt 薄膜是不连续的，正常厚度为 25 埃。在室温下，抗 SEE 与磷酸盐缓冲盐水 (PBS) 中的 SEE 结合期间，Pt 膜的阻抗降低。在特异性结合位点饱和之前，PBS 中的阻抗降低是不可逆的。饱和时，100 Hz 下的阻抗是 PES 中新鲜抗 SEE 层获得的值的 14%。阻抗 (/Z/) 降低的幅度与 0.389 至 10.70 ng/ml SEE 范围内的 SEE 浓度存在简单关系。当传感器暴露于 PBS 中的 100 ng/ml κ-酪蛋白、α-酪蛋白或 α-乳白蛋白时，没有发生不可逆的阻抗降低，从而证明了生物传感器的特异性。
Immunobiosensing techniques to measure specific antigen-antibody binding reactions are important in the development of biosensor applications in biotechnology, in vitro diagnosis, medicine and food technology.An immunobiosensor was constructed to measure the specific binding reaction between Staphylococcus enterotoxin B (SEE) and anti-SEE antibodies. The biosensor comprised an anti-SEE bioactive layer covalently immobilized on an ultra-thin platinum (Pt) film sputtered onto a 100 nm thick silicon dioxide layer on a silicon chip. The Pt film was discontinuous with a normal thickness of 25 Angstrom. The impedance of the Pt film decreased during the binding of the anti-SEE to SEE in phosphate buffered saline (PBS) at room temperature. The impedance decreases were irreversible in PBS before saturation of the specific binding sites. When saturated, the impedance at 100 Hz was 14% of the value obtained for the fresh anti-SEE layer in PES. The magnitude of the impedance (/Z/) decrease followed a simple relationship with SEE concentration in the range between 0.389 and 10.70 ng/ml SEE. The specificity of the biosensor was demonstrated by showing that no irreversible impedance decreases occurred when the sensor was exposed to 100 ng/ml kappa-casein, alpha-casein, or alpha-lactalbumin, in PBS.