Immunoengineered magnetic-quantum dot nanobead system for the isolation and detection of circulating tumor cells

Abstract

Background Highly efficient capture and detection of circulating tumor cells (CTCs) remain elusive mainly because of their extremely low concentration in patients’ peripheral blood. Methods We present an approach for the simultaneous capturing, isolation, and detection of CTCs using an immuno-fluorescent magnetic nanobead system (iFMNS) coated with a monoclonal anti-EpCAM antibody. Results The developed antibody nanobead system allows magnetic isolation and fluorescent-based quantification of CTCs. The expression of EpCAM on the surface of captured CTCs could be directly visualized without additional immune-fluorescent labeling. Our approach is shown to result in a 70–95% capture efficiency of CTCs, and 95% of the captured cells remain viable. Using our approach, the isolated cells could be directly used for culture, reverse transcription-polymerase chain reaction (RT-PCR), and immunocytochemistry (ICC) identification. We applied iFMNS for testing CTCs in peripheral blood samples from a lung cancer patient. Conclusions It is suggested that our iFMNS approach would be a promising tool for CTCs enrichment and detection in one step. Graphic abstract

Dual-Aptamer-Targeted Immunomagnetic Nanoparticles to Accurately Explore the Correlations between Circulating Tumor Cells and Gastric Cancer

“…For the preparation of streptavidin (SA)-modified Fe 3 O 4 nanoparticles (MNs), 100 μL of 1-ethyl-3-(3-dimethylaminopropyl)­carbodiimide (EDC) (10 mg/mL) and 100 μL of N -hydroxysuccinimide (NHS) (10 mg/mL), which were dissolved in MEST (100 mM MES, pH 5.0, 0.05% Tween 20), were added to 1.0 mL of MNs at 25 °C for 30 min to activate the carboxyl groups on the surface of MNs. Then, 100 μL of SA (1.0 mg/mL) was added and incubated at 25 °C for 1 h. Afterward, the supernatant was removed by magnetic separation, and 200 μL of 1% bovine serum albumin (BSA) containing PBST was added to seal the unreacted activated carboxyl groups on the surface of MNs . After washing with PBS, the product was stored in PBS at 4 °C.…”

“…Then, 100 μL of SA (1.0 mg/mL) was added and incubated at 25 °C for 1 h. Afterward, the supernatant was removed by magnetic separation, and 200 μL of 1% bovine serum albumin (BSA) containing PBST was added to seal the unreacted activated carboxyl groups on the surface of MNs. 43 After washing with PBS, the product was stored in PBS at 4 °C. Preparation of Dual-Aptamer-Modified Fe 3 O 4 Immunomagnetic Particles (IMNs).…”

Metal Nanoparticles as Novel Agents for Lung Cancer Diagnosis and Therapy

“…Zhang et al developed an immunofluorescent magnetic nanobead system (iFMNS), and achieved CTCs enrichment and visualization in one step without additional immune‐fluorescent labeling. [ 76 ] This FMN nanoprobe (114 nm) was coated with a monoclonal antibody against EpCAM (i.e., CD326, which is highly expressed in epithelial tumor tissues) ( Figure a). About 70–95% of the cancer cells could be isolated by FMN nanoprobe (encapsulated with numerous CdSe/ZnS QDs) within 10 min under an external magnetic field.…”

“…EpCAM antigen (red) was potentially useful for CTCs identification. Reproduced with permission [76]. Copyright 2020, BMC.…”

Polycarboxybetaine-Based Hydrogels for the Capture and Release of Circulating Tumor Cells

“…One of the common biomarkers for the identification of CTCs is the epithelial cell adhesion molecule (EpCAM), a 30–40 kDa glycosylated type I membrane protein [ 3 , 4 ], which is frequently overexpressed in cancer cell membranes, but is absent from hematologic cells, making the protein a good candidate for use in the efficient isolation of CTCs from blood. Therefore, anti-EpCAM antibodies have been utilized in devices for the detection and capture of CTCs [ 5 , 6 , 7 , 8 , 9 ].…”