High-Efficiency Isolation and Rapid Identification of Heterogeneous Circulating Tumor Cells (CTCs) Using Dual-Antibody-Modified Fluorescent-Magnetic Nanoparticles

Abstract

Extreme rarity and inherent heterogeneity of circulating tumor cells (CTCs) result in a tremendous challenge for the CTC isolation from patient blood samples with high efficiency and purity. Current CTC isolation approaches mainly rely on the epithelial cell adhesion molecule (EpCAM), which may significantly reduce the ability to capture CTCs when the expression of EpCAM is lost or down-regulated in epithelial–mesenchymal transition. Here, a rapid and highly efficient method is developed to isolate and identify heterogeneous CTCs with high efficiency from patient blood samples using the fluorescent-magnetic nanoparticles (F-MNPs). A dual-antibody interface targeting EpCAM and N-cadherin is fabricated onto the F-MNPs to capture epithelial CTCs as well as mesenchymal CTCs from whole blood samples. The poly­(carboxybetaine methacrylate) brushes of excellent antifouling properties are employed to decrease nonspecific cell adhesion. Moreover, the F-MNPs provide a prompt identification strategy for heterogeneous CTCs (F-MNPs+, Hoechst 33342+, and CD45−) that can directly identify CTCs in a gentle one-step processing within 1 h after isolation from patient blood samples. This has been demonstrated through artificial samples as well as patient samples in details.

Cell‐Released Magnetic Vesicles Capturing Metabolic Labeled Rare Circulating Tumor Cells Based on Bioorthogonal Chemistry

“…[13] To cope with the challenge, immunomagnetic bead separation methods have emerged as a promising class of strategy to specifically capture CTCs through a rapid and magnetic responsive way. [14][15][16][17] In order to satisfy the ultimate purpose of CTCs enrichment with high purity and high efficient, two essential terms, the antifouling surface and immobilized monoclonal-antibodies/aptamers, are particularly required in design and fabrication of the sophisticated magnetic capture platforms. More specifically, the antifouling surface eliminates non-specific adsorption of non-CTCs or proteins in the CTCs recognition and binding stage, [18][19][20] and the monoclonal-antibodies/aptamers provide an immunoaffinity channel to recognize/bind specific antigens, complement components, and receptors on CTCs membranes.…”

“…In recent years, our group and other researchers have attempted to reduce the influence of cellular heterogeneity in CTCs enrichment by incorporating multi-targeting ligands (anti-EpCAM, folic acid, hyaluronic acid) [25] or antibody-cocktail strategy [16,26,27] in the magnetic bead systems, thus CTCs with known surface markers/receptors are still basically required. Therefore, a way that can generate a type of "artificial target" on all tumor cells might help us get out of this dilemma.…”

Bioinspired DNA Nanointerface with Anisotropic Aptamers for Accurate Capture of Circulating Tumor Cells

“…To address the above issue, many methods based on cocktail of various capture ligand had been exploited for the accurate capture of CTCs. [24,25] To date, the successful improvement of capturing heterogeneous CTCs via cocktail of aptamers was achieved by decorating different aptamers on the same substrates. [26,27] However, the conformation of aptamers would be affected by the interaction between substrates and aptamers, leading to limited capture performance.…”

Microfluidics and Nanomaterial-based Technologies for Circulating Tumor Cell Isolation and Detection

“…The F-MNPs are fabricated to conjugate a dual-antibody interface targeting EpCAM and N-cadherin to capture epithelial CTCs and mesenchymal CTCs from whole blood samples. The F-MNPs were validated in CTC isolation and CTC identification in the blood samples of cancer patients [103] ( Figure 7 E-I).…”