Abstract:
Traumatic brain injury (TBI) is a sudden brain injury due to an external force that causes a large number of deaths and permanent disabilities every year. S100B has been recognized as a potential objective quantitative biomarker for screening the prognosis of TBI and severe head injury. In this article, an anti-S100B monoclonal antibody was immobilized on cysteamine (Cy) functionalized gold nanoparticles (AuNPs) by EDC-NHS chemistry, which enabled S100B resonance Rayleigh scattering (RRS) detection based on antibody-labeled gold nanoparticles. The prepared conjugates were characterized by ultraviolet-visible spectroscopy (UV-vis), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), dynamic light scattering (DLS) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Based on the specific binding of the antibody and antigen, the RRS intensities at 381 nm and 541 nm wavelengths were significantly enhanced, and thus a dual wavelength overlapping resonance Rayleigh scattering (DWO-RRS) method was established. The scattering intensity of the two overlapping peaks was proportional to the concentration of S100B in the range of 0.05-4.5 ng mL-1 with a detection limit of 0.002 ng mL-1. The proposed DWO-RRS method is time-saving, simple, sensitive, and can be used to determine the concentration of S100B in human serum with satisfactory results, which has a promising application in the early diagnosis of TBI.
摘要:
创伤性脑损伤(TBI)是由于外力引起的突发性脑损伤,每年导致大量死亡和永久性残疾。S100B已被认为是筛选TBI和重型颅脑损伤预后的潜在客观定量生物标志物。在这篇文章中,通过EDC-NHS化学将抗S100B单克隆抗体固定在半胱胺(Cy)功能化的金纳米颗粒(AuNPs)上,这使得基于抗体标记的金纳米粒子的S100B共振瑞利散射(RRS)检测成为可能。用紫外-可见光谱(UV-vis)对所制备的偶联物进行了表征,透射电子显微镜(TEM),傅里叶变换红外光谱(FTIR),动态光散射(DLS)和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)。基于抗体和抗原的特异性结合,381nm和541nm波长处的RRS强度显著增强,建立了双波长重叠共振瑞利散射(DWO-RRS)方法。两个重叠峰的散射强度与S100B的浓度成正比,范围为0.05-4.5ngmL-1,检出限为0.002ngmL-1。提出的DWO-RRS方法节省时间,简单,敏感,并可用于测定人血清中S100B的浓度,结果满意,在TBI的早期诊断中具有广阔的应用前景。