Rapid Electrochemical Assessment of Tumor Suppressor Gene Methylations in Raw Human Serum and Tumor Cells and Tissues Using Immunomagnetic Beads and Selective DNA Hybridization

We report a rapid and sensitive electrochemical strategy for the detection of gene‐specific 5‐methylcytosine DNA methylation. Magnetic beads (MBs) modified with an antibody for 5‐methylcytosines (5‐mC) are used for the capture of any 5‐mC methylated single‐stranded (ss)DNA sequence. A flanking regio...

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Bibliographic Details
Published in:Angewandte Chemie International Edition 2018-07, Vol.57 (27), p.8194-8198
Main Authors: Povedano, Eloy, Valverde, Alejandro, Montiel, Víctor Ruiz‐Valdepeñas, Pedrero, María, Yáñez‐Sedeño, Paloma, Barderas, Rodrigo, San Segundo‐Acosta, Pablo, Peláez‐García, Alberto, Mendiola, Marta, Hardisson, David, Campuzano, Susana, Pingarrón, José M.
Format: Article
Language:English
Subjects:
Beads
biofluids
Biosensors
Brain
brain tissues
Brain tumors
Deoxyribonucleic acid
DNA
DNA methylation
DNA methyltransferase
dual detection
Electrical measurement
Electrochemistry
Glioblastoma
Glioblastoma cells
Hybridization
immuno-DNA sensors
methylated DNA
Methylguanine
Nucleotide sequence
O6-methylguanine-DNA methyltransferase
Paraffin
Pretreatment
Tissues
Tumor cells
Tumor suppressor genes
Tumors
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Summary:We report a rapid and sensitive electrochemical strategy for the detection of gene‐specific 5‐methylcytosine DNA methylation. Magnetic beads (MBs) modified with an antibody for 5‐methylcytosines (5‐mC) are used for the capture of any 5‐mC methylated single‐stranded (ss)DNA sequence. A flanking region next to the 5‐mCs of the captured methylated ssDNA is recognized by hybridization with a synthetic biotinylated DNA sequence. Amperometric transduction at disposable screen‐printed carbon electrodes (SPCEs) is employed. The developed biosensor has a dynamic range from 3.9 to 500 pm and a limit of detection of 1.2 pm for the methylated synthetic sequence of the tumor suppressor gene O‐6‐methylguanine‐DNA methyltransferase (MGMT) promoter region. The method is applied in the 45‐min analysis of specific methylation in the MGMT promoter region directly in raw spiked human serum samples and in genomic DNA extracted from U‐87 glioblastoma cells and paraffin‐embedded brain tumor tissues without any amplification and pretreatment step. An immuno‐DNA electrochemical sensor for the rapid assessment of tumor suppressor gene methylations in raw human serum, tumor cells, and cancer tissues is presented.
ISSN:1433-7851
1521-3773
DOI:10.1002/anie.201804339