Székács András, Adányi Nóra, Székács Inna, Majer-Baranyi Krisztina, Szendro István
Plant Protection Institute, Hungarian Academy of Sciences, POB 102, H-1525 Budapest, Hungary.
Appl Opt. 2009 Feb 1;48(4):B151-8. doi: 10.1364/ao.48.00b151.
Coupling the high specificity of the immunoanalytical reaction with the high sensitivity of optical waveguide light-mode spectroscopy (OWLS) detection gives the possibility to develop immunosensors with in most cases a definitely lower detection limit than traditionally used immunoassays. Measurements were performed on the sensitized surface of optical waveguide grating coupler sensors (2400 lines/mm grating). The OWLS technique is based on the precise measurement of the resonance angle of a polarized laser light (632.8 nm), diffracted by a grating and incoupled into a thin waveguide. The effective refractive index, determined from the resonance incoupling angle detected at high accuracy, allows determination of layer thickness and coverage (or mass) of the adsorbed or bound material with ultrahigh sensitivity. OWLS immunosensors were developed as label-free immunosensors with an amino group modified SiO(2)-TiO(2) sensor surface on which the immunoreactants could be anchored. One of the components of the antibody-antigen complex was chemically bound on the sensor surface, allowing noncompetitive or competitive detection of the analytes. To illustrate that the resulting immunosensors are suitable for the determination of small and large molecular weight analytes, OWLS sensor formats were applied for quantitative detection of a herbicide active ingredient trifluralin, a Fusarium mycotoxin zearalenone, and an egg yolk protein of key importance in endocrine regulation, vitellogenin.
将免疫分析反应的高特异性与光波导光模式光谱(OWLS)检测的高灵敏度相结合,使得开发免疫传感器成为可能,在大多数情况下,其检测限肯定低于传统使用的免疫分析方法。测量是在光波导光栅耦合器传感器(2400线/毫米光栅)的敏化表面上进行的。OWLS技术基于对偏振激光(632.8纳米)共振角的精确测量,该激光由光栅衍射并耦合到薄波导中。通过高精度检测到的共振耦合角确定的有效折射率,能够以超高灵敏度测定吸附或结合物质的层厚度和覆盖率(或质量)。OWLS免疫传感器被开发为无标记免疫传感器,其具有氨基修饰的SiO(2)-TiO(2)传感器表面,免疫反应物可固定在该表面上。抗体 - 抗原复合物的其中一个组分被化学结合在传感器表面,从而实现对分析物的非竞争性或竞争性检测。为了说明所得的免疫传感器适用于测定小分子和大分子重量的分析物,OWLS传感器形式被应用于定量检测除草剂活性成分氟乐灵、镰刀菌霉菌毒素玉米赤霉烯酮以及在内分泌调节中至关重要的蛋黄蛋白卵黄蛋白原。