[Application of optical proteinchip in detecting phage M13KO7].
[Application of optical proteinchip in detecting phage M13KO7].
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DOI：
10.1016/s1872-2075(06)60060-5
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发表时间：
2006-09
期刊：
影响因子：
--
通讯作者：
Jin G
中科院分区：
文献类型：
--
作者：
Qi C;Feng J;Wang ZH;Meng YH;Yan XY;Jin G
在醛修饰的硅片表面结合亲和素层。利用亲和素与生物素的相互作用，将鼠单克隆生物素-抗M13（抗体GP 3）标记的生物素固定化。将表面在含有噬菌体M13 KO 7的溶液中孵育，所述噬菌体M13 KO 7通过噬菌体M13 KO 7和抗体GP 3之间的相互作用被抗体GP 3捕获，导致通过成像椭圆偏振法检测的层厚度的变化。结果显示在硅片表面上具有厚度约6.9nm的抗体GP 3的饱和层。噬菌体M13 KO 7与抗体GP 3之间的特异性相互作用导致层厚度的变化。当抗体浓度为1.1× 10 ~（10）pfu/mL时，噬菌体M13 KO 7与抗体GP 3的结合层为17.5nm。每一个层厚度的变化对应于噬菌体M13 KO 7的浓度在0.1×1010-2.5×1010 pfu/mL范围内，灵敏度为109 pfu/mL。与其他方法相比，光学蛋白质芯片具有测量时间短、无标记、定量、可视化等优点。本研究对研究抗体与病毒的相互作用具有重要意义，在病毒病的早期诊断中具有潜在的应用价值。
Avidin layer was bound on the substrate surface of Silicon wafer modified with aldehyde. The interaction between avidin and biotin was adopted for the immobilization of mouse monoclonal biotin-anti-M13 (antibody GP3)-labeled biotin. The surface was incubated in a solution containing phage M13KO7, which was trapped by the antibody GP3 with the interaction between phage M13KO7 and antibody GP3, resulting in a variation of layer thickness that was detected by imaging ellipsometry. The results showed a saturated layer of antibody GP3 with a thickness about 6.9 nm on the surface of the silicon wafer. The specific interaction between phage M13KO7 and antibody GP3 resulted in a variation of layer thickness. The layer of phage M13KO7 bound with antibody GP3 was 17.5 nm in the concentration of 1.1×1010 pfu/mL. Each variation of the layer thickness corresponded to a concentration of phage M13KO7 in the range of 0.1×1010–2.5×1010 pfu/mL, with the sensitivity of 109 pfu/mL. Compared with other methods, the optical protein-chip, requiring only short measurement time, label free, is a quantitative test, and can be visualized. This study could be significant on the interactions between the antibody and the virus, showing potential in the early diagnosis of virosis.