Integrating electrochemical immunosensing and cell adhesion technologies for cancer cell detection and enumeration

Abstract

We have successfully integrated techniques for controlling cell adhesion and performing electrochemical differential pulse voltammetry (DPV) through the use of digitally controlled microfluidics and patterned transparent indium tin oxide electrode arrays to enable rapid and sensitive enumeration of cancer cells in a scalable microscale format. This integrated approach leverages a dual-working electrode (WE) surface to improve the specificity of the detection system. Here, one of the WE surfaces is functionalized with anti-Melanocortin 1 Receptor antibodies specific to melanoma cancer cells, while the other WE acts as a control (i.e., without antibody), for detecting non-specific interactions between cells and the electrode. The method is described and shown to provide effective detection of melanoma cells at concentrations ranging between 25 to 300 cells per 20 μL sample volume after a 5 min incubation and 15 s of DPV measurements. The estimated limit of detection was ~17 cells. The sensitivity and specificity of the assay were quantified using addition of large fractions of non-target cells and resulted in a detection reproducibility of ~97%. The proposed approach demonstrates a unique integration of electrochemical sensing and microfluidic cell adhesion technologies with multiple advantages such as label-free detection, short detection times, and low sample volumes. Next steps for this platform include testing with patient samples and use of other cell-surface biomarkers for detection and enumeration of circulating tumor cells in prostate, breast, and colon cancer.

Review—Measurement and Analysis of Cancer Biomarkers Based on Electrochemical Biosensors

“…A wide LR from 10 to 10 6 cells mL −1 with a detection limit as low as 10 cells mL −1 was achieved. Seenivasan et al 159 used a dual-working electrode (WE) surface to improve the specificity of the detection system. One of the WE surface is functionalized with anti-Melanocortin 1 Receptor antibodies specific to melanoma cancer cells, while the other WE act as a control (i.e., without antibody), for detecting non-specific interactions between cells and the electrode.…”

Recent Development of Nanomaterials-Based Cytosensors for the Detection of Circulating Tumor Cells

“…Thus, noble metal NPs have been intensively used to construct electrochemical biosensors [142][143][144]. Seenivasan et al employed anti-MC1R-Ab-labeled AuNPs to detect CTCs by differential pulse voltammetry (DPV) [145]. Au nanowire arrays were prepared by electrochemical deposition with anodic aluminum oxide as template for the capture and release of CCRF-CEM cells [146].…”

Clinical Relevance of Liquid Biopsy in Melanoma and Merkel Cell Carcinoma

“…In parallel, many other new technologies have been developed to overcome the challenge of CMC detection. Different microfluidic chips and biosensors have been tested in patients with melanoma [34][35][36][37][38][39][40]; however, the multiplicity of procedures reduces their potential clinical utility, like previously observed for RT-PCR-based methods.…”