Regiart Matías, Fernández-Baldo Martin A, Villarroel-Rocha Jhonny, Messina Germán A, Bertolino Franco A, Sapag Karim, Timperman Aaron T, Raba Julio
INFAP, Laboratorio de Sólidos Porosos, Universidad Nacional de San Luis, CONICET, Ejercito de los Andes 950, D5700BWS, San Luis, Argentina.
INQUISAL, Departamento de Química, Universidad Nacional de San Luis, CONICET, Chacabuco 917, D5700BWS, San Luis, Argentina.
Anal Chim Acta. 2017 Apr 22;963:83-92. doi: 10.1016/j.aca.2017.01.029. Epub 2017 Jan 30.
We report a hybrid glass-poly (dimethylsiloxane) microfluidic immunosensor for epidermal growth factor receptor (EGFR) determination, based on the covalent immobilization of anti-EGFR antibody (anti-EGFR) on amino-functionalized mesoporous silica (AMS) retained in the central channel of a microfluidic device. The synthetized AMS was characterized by N adsorption-desorption isotherm, scanning electron microscopy (SEM), energy dispersive spectrometry (EDS) and infrared spectroscopy. The cancer biomarker was quantified in human serum samples by a direct sandwich immunoassay measuring through a horseradish peroxidase-conjugated anti-EGFR. The enzymatic product was detected at -100 mV by amperometry on a sputtering gold electrode, modified with an ordered mesoporous carbon (CMK-3) in a matrix of poly-acrylamide-co-methacrylate of dihydrolipoic acid (poly(AC-co-MDHLA)) through in situ copolymerization. CMK-3/poly(AC-co-MDHLA)/gold was characterized by cyclic voltammetry, EDS and SEM. The measured current was directly proportional to the level of EGFR in human serum samples. The linear range was from 0.01 ng mL to 50 ng mL. The detection limit was 3.03 pg mL, and the within- and between-assay coefficients of variation were below 5.20%. The microfluidic immunosensor is a very promising device for the diagnosis of several kinds of epithelial origin carcinomas.
我们报道了一种用于检测表皮生长因子受体（EGFR）的混合玻璃 - 聚二甲基硅氧烷微流控免疫传感器，其基于抗EGFR抗体（anti - EGFR）共价固定在保留于微流控装置中央通道的氨基功能化介孔二氧化硅（AMS）上。合成的AMS通过N吸附 - 解吸等温线、扫描电子显微镜（SEM）、能量色散光谱（EDS）和红外光谱进行表征。通过辣根过氧化物酶偶联的抗EGFR进行直接夹心免疫测定，对人血清样本中的癌症生物标志物进行定量。在通过原位共聚在聚 - 丙烯酰胺 - 二氢硫辛酸甲基丙烯酸酯（poly(AC - co - MDHLA)）基质中修饰有有序介孔碳（CMK - 3）的溅射金电极上，通过安培法在 - 100 mV下检测酶产物。通过循环伏安法、EDS和SEM对CMK - 3/poly(AC - co - MDHLA)/金进行表征。所测电流与人血清样本中EGFR水平成正比。线性范围为0.01 ng/mL至50 ng/mL。检测限为3.03 pg/mL，批内和批间变异系数均低于5.20%。该微流控免疫传感器是一种用于诊断多种上皮源性癌的非常有前景的装置。
