Magnetic Nanowire Networks for Dual-Isolation and Detection of Tumor-Associated Circulating Biomarkers

Abstract

Purpose: Recent developments in genomic and molecular methods have revolutionized the range of utilities of tumor-associated circulating biomarkers in both basic and clinical research. Herein, we present a novel approach for ultrasensitive extraction of cfDNA and CTCs, at high yield and purity, via the formation of magnetic nanowire networks.Materials and Methods: We fabricated and characterized biotinylated cationic polyethylenimine and biotinylated antibody cocktail-conjugated magnetic polypyrrole NWs (PEI/mPpy NW and Ab cocktail/mPpy NW, respectively). We applied these NWs to the extraction of cfDNA and CTC from the blood of 14 patients with lung cancer. We demonstrated reliable detection of EGFR mutations based on digital droplet PCR analysis of cfDNA and CTC DNA from patients with lung cancer.Results: The NW networks confined with a high density of magnetic nanoparticles exhibited superior saturation magnetization, which enabled rapid and high-yield capture whilst avoiding or minimizing damage and loss. The NW networks enabled the co-isolation of CTCs and cfDNA of high quality and sufficient quantities, thus allowing the amplification of rare and low-prevalence cancer-related mutations.Conclusion: The simple, versatile, and highly efficient nanowire network tool allows sensitive and robust assessment of clinical samples.

Projection method as a probe for multiplexing/demultiplexing of magnetically enriched biological tissues

“…31,32 Furthermore, the MNWs have shown a high internalization by cells in comparison to other magnetic nanoparticles, such as IONs, improving the enrichment and multiplexing yield. 26,28,33 As a consequence, MNWs have recently attracted attention as enrichment agents for studying breast and lung cancers, 34,35 CTCs, 36 and leukemia. 37 Regardless of the tremendous opportunities for MNWs, they have not been used as self-standing biomarkers for multiplexing applications up to date due to three major drawbacks.…”

The Growing Impact of Micro/Nanomaterial‐Based Systems in Precision Oncology: Translating “Multiomics” Technologies

“…[36] The integrated nanomaterial enabled the maximal loading of capture agents and magnetic particles for superior capture and purification of cfDNA at 0.01-1000 ng mL −1 . [36] Toralla et al developed an integrated microfluidic magnetic extraction platform (METRO) for cfDNA isolation from biological samples such as undiluted serum. [37] The capture domain of the microfluidic platform was equipped with paramagnetic microparticles which can be expanded by flowing binding buffer (pH < 6) through the channel.…”

“…Reproduced with permission. [36] Copyright 2018, Ivyspring. manipulation of DNA under an electric field.…”

One-Step Polymerase Chain Reaction-Free Nanowire-Based Plasma Cell-Free DNA Assay to Detect EML4-ALK Fusion and to Monitor Resistance in Lung Cancer

“…Recently, we developed a new genotyping assay that accurately and rapidly detects small amounts of tumor-derived cell-free DNA (cfDNA) in various body fluids in patients with lung cancer or cervical cancer. [18][19][20] This assay isolates cfDNA in body fluids using nanowires (NW) and directly detects DNA variants or mutations without PCR amplification within 1 hour. Our previous studies demonstrated that this NW-based cfDNA assay can be used to detect low-abundance DNA variants in plasma cfDNA with high sensitivity and specificity.…”

“…In particular, due to the vast amount of raw data generated by NGS sequencing and its complexity, the role of bioinformatics experts is crucial for processing, interpreting, and analyzing complex bioinformatics data, ultimately obtaining biological specificity from sequenced samples. Recently, we developed a new genotyping assay that accurately and rapidly detects small amounts of tumor‐derived cell‐free DNA (cfDNA) in various body fluids in patients with lung cancer or cervical cancer [18–20]. This assay isolates cfDNA in body fluids using nanowires (NWs) and directly detects DNA variants or mutations without PCR amplification within 1 hour.…”