Ankara University, Department of Chemistry, Science Faculty, Tandoğan, Ankara 06100, Turkey.
Biosens Bioelectron. 2013 Aug 15;46:53-60. doi: 10.1016/j.bios.2013.01.060. Epub 2013 Feb 19.
A novel highly sensitive impedimetric Myelin Basic Protein (MBP) immunosensor for the determination of a Multiple Sclerosis (MS) autoantibody, Anti-Myelin Basic Protein (Anti-MBP) was developed by immobilization of MBP on Gelatin and Gelatin-Titanium Dioxide (TiO₂) modified platinium electrode. Cyclic voltammetric (CV) and Electrochemical Impedance Spectroscopic (EIS) methods were employed in determination of the electrode responses and applicability. Gelatin-MBP and gelatin-TiO₂-MBP electrodes were prepared by chemical immobilization of the substrates onto the platinium electrodes. The formal potentials of MBP confined on gelatin-MBP and gelatin-TiO₂-MBP surfaces are estimated to be 195 and 205 mV, respectively. Thus, a little more reversible electron transfer reaction occurs on the gelatin-TiO₂-MBP immunosensor surface. The peak separations of MBP (150 mV and 110 mV s(-1) at 100 mV s(-1)) and the asymmetric anodic and cathodic peak currents indicate that the electron transfer between Anti-MBP and gelatin-MBP/gelatin-TiO₂-MBP immunosensor is quasireversible. Control samples containing a nonspecific human immunoglobulin G (hIgG) antibody were also studied, and calibration curves were obtained by subtraction of the responses for specific and nonspecific antibody-based sensors. Gelatin-MBP and gelatin-TiO₂-MBP immunosensors have detection limit of 0.1528 ng ml(-1) and 0.1495 ng ml(-1) respectively. This immunosensor exhibits high sensitivity and low response times (58 s for gelatin-MBP and 46 s for gelatin-TiO₂-MBP immunosensor). The developed label-free impedimetric immunosensors also provide a simple and sensitive detection method for the specific determination of Anti-MBP in human cerebrospinal fluid (CSF) and serum samples.
一种新型的高灵敏度基于免疫测定的髓鞘碱性蛋白（MBP）免疫传感器，用于测定多发性硬化症（MS）自身抗体，抗髓鞘碱性蛋白（Anti-MBP），是通过将 MBP 固定在明胶和明胶-二氧化钛（TiO₂）修饰的铂电极上来制备的。采用循环伏安法（CV）和电化学阻抗谱（EIS）方法来确定电极的响应和适用性。通过将底物化学固定在铂电极上来制备明胶-MBP 和明胶-TiO₂-MBP 电极。MBP 固定在明胶-MBP 和明胶-TiO₂-MBP 表面的形式电位分别估计为 195 和 205 mV，因此，在明胶-TiO₂-MBP 免疫传感器表面上发生的电子转移反应稍微更可逆。MBP 的峰分离（在 100 mV s(-1) 时为 150 mV 和 110 mV s(-1)）和不对称的阳极和阴极峰电流表明，Anti-MBP 与明胶-MBP/明胶-TiO₂-MBP 免疫传感器之间的电子转移是准可逆的。还研究了含有非特异性人免疫球蛋白 G（hIgG）抗体的对照样品，并通过从基于特异性和非特异性抗体的传感器的响应中减去来获得校准曲线。明胶-MBP 和明胶-TiO₂-MBP 免疫传感器的检测限分别为 0.1528 ng ml(-1)和 0.1495 ng ml(-1)。该免疫传感器具有高灵敏度和低响应时间（明胶-MBP 为 58 s，明胶-TiO₂-MBP 为 46 s）。开发的无标记阻抗免疫传感器还为在人脑脊液（CSF）和血清样品中特异性测定 Anti-MBP 提供了一种简单而灵敏的检测方法。
