Abstract
Ligand-directed targeting and capturing of cancer cells is a new approach for detecting circulating tumor cells (CTCs). Ligands such as antibodies have been successfully used for capturing cancer cells and an antibody based system (CellSearch®) is currently used clinically to enumerate CTCs. Here we report the use of a peptide moiety in conjunction with a microcantilever array system to selectively detect CTCs resulting from cancer, specifically breast cancer. A sensing microcantilever, functionalized with a breast cancer specific peptide 18-4 (WxEAAYQrFL), showed significant deflection on cancer cell (MCF7 and MDA-MB-231) binding compared to when exposed to noncancerous (MCF10A and HUVEC) cells. The peptide-functionalized microcantilever allowed efficient capture and detection of cancer cells in MCF7 spiked human blood samples emulating CTCs in human blood. A detection limit of 50–100 cancer cells mL−1 from blood samples was achieved with a capture yield of 80% from spiked whole blood samples. The results emphasize the potential of peptide 18-4 as a novel peptide for capturing and detecting cancer cells in conjunction with nanomechanical cantilever platform. The reported peptide-based cantilever platform represents a new analytical approach that can lead to an alternative to the various detection platforms and can be leveraged to further study CTCs.
Highlights
In-vivo examinations of breast cancer is mainly implemented through techniques like mammography, ultrasound exams, magnetic resonance imaging (MRI) and/or [18F]fluorodeoxyglucose positron emission tomography, which are typically followed by ex vivo biopsy and further checkups[1]
® immunomagnetic beads conjugated with an antibody to epithelial cell adhesion molecule (EpCAM) (CellSearch, VeridexTM, Warren, PA), is clinically used for enumeration of circulating tumor cells (CTCs) from human blood samples[19]
We report the development of a peptide-based microcantilever array sensor for efficient capture of intact representative cancer cells at low concentrations without pre-requisite labeling or sample processing (Fig. 1)
Summary
In-vivo examinations of breast cancer is mainly implemented through techniques like mammography (an x-ray of the breast), ultrasound exams, magnetic resonance imaging (MRI) and/or [18F]fluorodeoxyglucose positron emission tomography, which are typically followed by ex vivo biopsy and further checkups[1]. Molecular and clinical findings have revealed that cancer cells may invade into the blood circulation at early stages of tumor development, emphasizing the importance of sensitive and specific detection of CTCs in the blood[1]. ® immunomagnetic beads conjugated with an antibody to EpCAM (CellSearch , VeridexTM, Warren, PA), is clinically used for enumeration of CTCs from human blood samples[19]. Majority of these advanced detection platforms rely on antibody and/or oligonucleotide probes for recognition, identification, and quantification of the target cells. We report the development of a peptide-based microcantilever array sensor for efficient capture of intact representative cancer cells at low concentrations without pre-requisite labeling or sample processing (Fig. 1). The cancer cells were detected by recording the nanomechanical bending of the cantilevers in real-time based on the surface stress induced by adhesion of the cancer cells to the immobilized peptides
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