Microfluidic immunosensor based on mesoporous silica platform and CMK-3/poly-acrylamide-co-methacrylate of dihydrolipoic acid modified gold electrode for cancer biomarker detection.
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 N2adsorption-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-1to 50 ng mL-1. The detection limit was 3.03 pg mL-1, 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.
INFAP, Laboratorio de Sólidos Porosos, Universidad Nacional de San Luis, CONICET, Ejercito de los Andes 950, D5700BWS, San Luis, Argentina.
Matías
M
Regiart
INQUISAL, Departamento de Química, Universidad Nacional de San Luis, CONICET, Chacabuco 917, D5700BWS, San Luis, Argentina.
Martin A
MA
Fernández-Baldo
INFAP, Laboratorio de Sólidos Porosos, Universidad Nacional de San Luis, CONICET, Ejercito de los Andes 950, D5700BWS, San Luis, Argentina.
Jhonny
J
Villarroel-Rocha
INQUISAL, Departamento de Química, Universidad Nacional de San Luis, CONICET, Chacabuco 917, D5700BWS, San Luis, Argentina.
Germán A
GA
Messina
INQUISAL, Departamento de Química, Universidad Nacional de San Luis, CONICET, Chacabuco 917, D5700BWS, San Luis, Argentina.
Franco A
FA
Bertolino
INFAP, Laboratorio de Sólidos Porosos, Universidad Nacional de San Luis, CONICET, Ejercito de los Andes 950, D5700BWS, San Luis, Argentina.
Karim
K
Sapag
Advanced Diagnostics & Therapeutics, Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA. Electronic address: atimperm@nd.edu.
Aaron T
AT
Timperman
INQUISAL, Departamento de Química, Universidad Nacional de San Luis, CONICET, Chacabuco 917, D5700BWS, San Luis, Argentina. Electronic address: jraba@unsl.edu.ar.
Julio
J
Raba
D000178:Acrylamides / Q000737:chemistry*; D014408:Biomarkers, Tumor / Q000032:analysis* / Q000097:blood; D004566:Electrodes; D006046:Gold / Q000737:chemistry*; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation*; D056656:Lab-On-A-Chip Devices; D058105:Polymerization; D011109:Polymethacrylic Acids / Q000737:chemistry*; D016062:Porosity; D012822:Silicon Dioxide / Q000737:chemistry*; D008063:Thioctic Acid / Q000031:analogs & derivatives* / Q000737:chemistry
D000178:Acrylamides; D014408:Biomarkers, Tumor; D011109:Polymethacrylic Acids; C049146:lymphostim; D008063:Thioctic Acid; D006046:Gold; D012822:Silicon Dioxide; C015144:dihydrolipoic acid
Cancer biomarker; Cancer diagnosis; EGFR; Mesoporous silica; Microfluidic immunosensor; Nanostructured electrode