摘要
建立了一种夹心式用于检测血清中甲胎蛋白(alpha-fetoprotein,AFP)的新型传感器。利用微孔板表面羧基与抗体表面的氨基之间的相互作用,将AFP的单克隆抗体(monoclonal antibodies,McAbs)修饰到微孔板的表面;再利用抗体抗原的定向固定效应,将AFP键合到McAbs的表面,接着将生物素标记的AFP单克隆抗体(biotinylated McAbs)与AFP结合;最后,利用链霉亲和素(streptavidin,SA)与生物素的特异性结合作用,将G-四链体-氯血红素DNA酶键合到生物素标记的单克隆抗体上,从而制得高灵度、高特异性的AFP传感器。用2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)-H2O2体系检测AFP的浓度,信号值与AFP的浓度在0.05 ng·mL-1至20 ng·mL-1之间成线性关系,检测限为0.02 ng·mL-1。此传感器可应用于血清中AFP的检测。
An ultrasensitive sandwich sensor for Alpha-fetoprotein （AFP） detection is introduced. The DNAzyme is tethered to biotinylated monoclonal antibodies （McAbs） which serves as the sensing element to recog- nize the target protein, and is then introduced onto the AFP-McAbs assembled microplate. The peroxidase-like DNAzyme catalyzes the oxidation of 2, 2＇ -azino-bis （3-ethylbenzothiazoline-6-sulfonic acid）, which generates a blue-green colorimetric signal. Under optimal conditions, the immunosensor exhibited a wide linear range from 0. 05 to 20 ng · mL-^ 1 relative to AFP concentration with a detection limit of 0. 02 ng · mL^- 1. This method can detect AFP in a serum-containing medium.
出处
《科学技术与工程》
北大核心
2014年第14期5-9,共5页
Science Technology and Engineering
基金
山东省高等学校科技计划项目(J10LB64)
山东省自然科学基金项目(2R2009BM003)资助
