High-Throughput Electrochemical Microfluidic Immunoarray for Multiplexed Detection of Cancer Biomarker Proteins

Abstract

Microchip-based microfluidic electrochemical arrays hold great promise for fast, high-throughput multiplexed detection of cancer biomarker proteins at low cost per assay using relatively simple instrumentation. Here we describe an inexpensive high-throughput electrochemical array featuring 32 individually addressable microelectrodes that is further multiplexed with an 8-port manifold to provide 256 sensors. The gold electrode arrays were fabricated by wet-etching commercial gold compact discs (CD-R) followed by patterned insulation. A print-and-peel method was used to create sub-microliter hydrophobic wells surrounding each sensor to eliminate cross contamination during immobilization of capture antibodies. High-throughput analyses were realized using eight 32-sensor immunoarrays connected to the miniaturized 8-port manifold, allowing 256 measurements in <1 h. This system was used to determine prostate cancer biomarker proteins prostate specific antigen (PSA), prostate specific membrane antigen (PSMA), interleukin-6 (IL-6), and platelet factor-4 (PF-4) in serum. Clinically relevant detection limits (0.05 to 2 pg mL−1) and 5-decade dynamic ranges (sub pg mL−1 to well above ng mL−1) were achieved for these proteins utilizing precapture of analyte proteins on magnetic nanoparticles decorated with enzyme labels and antibodies.

Microchip electrophoresis separation of a panel of preterm birth biomarkers

“…The field of integrated microfluidic biomarker analysis has been growing rapidly. Just in recent years, new microfluidic platforms for biomarker analysis have emerged with capabilities for colorimetric or electrochemical assays , ELISA , and other biosensors . However, a common disadvantage of most of these studies is that they were only capable of analyzing a single analyte per device.…”

“…However, a common disadvantage of most of these studies is that they were only capable of analyzing a single analyte per device. Tang et al.’s work was an exception where four cancer biomarkers were measured, but this analysis required off‐chip sample preparation and an array of sensors to achieve multiplexing.…”

Progress in Microfluidics‐Based Exosome Separation and Detection Technologies for Diagnostic Applications

“…In recent years, microfluidics‐based electrochemical biosensors have been favored by researchers. Many microfluidics‐based electrochemical sensors have successfully demonstrated that they have great potential for biomarker detection . These detection chips for various antibody proteins have great potential for detection of exosomes.…”

Magnetic nanoformulations for prostate cancer

“…All of these findings demonstrate PSMA conjugation benefits both tissue-specific detection/imaging and therapeutic outcomes [90]. A proof-of-concept study evaluating clinically relevant detection limits of PSMA (0.05 to 2 pg/mL) can be achieved through magnetic nanoparticle decorated with anti-PSMA in high-throughput electrochemical microfluidic immunoarray [89]. According to Table 2 and other related studies, it is important to note that PSMA ligand tagged formulation can be successfully employed as a multi-/dual-modality probe for prostate cancer imaging.…”