The past decade has witnessed ongoing progress in precision medicine to improve human health. As an emerging diagnostic technique, liquid biopsy can provide real-time, comprehensive, dynamic physiological and pathological information in a noninvasive manner, opening a new window for precision medicine. Liquid biopsy depends on the sensitive and reliable detection of circulating targets (e.g., cells, extracellular vesicles, proteins, microRNAs) from body fluids, the performance of which is largely governed by recognition ligands. Aptamers are single-stranded functional oligonucleotides, capable of folding into unique tertiary structures to bind to their targets with superior specificity and affinity. Their mature evolution procedure, facile modification, and affinity regulation, as well as versatile structural design and engineering, make aptamers ideal recognition ligands for liquid biopsy. In this review, we present a broad overview of aptamer-based liquid biopsy techniques for precision medicine. We begin with recent advances in aptamer selection, followed by a summary of state-of-the-art strategies for multivalent aptamer assembly and aptamer interface modification. We will further describe aptamer-based micro-/nanoisolation platforms, aptamer-enabled release methods, and aptamer-assisted signal amplification and detection strategies. Finally, we present our perspectives regarding the opportunities and challenges of aptamer-based liquid biopsy for precision medicine.

Aptamer-Based Detection of Circulating Targets for Precision Medicine.

Emerging 2D nanomaterials for biomedical applications

In recent years, an increasing number of nanomaterials have been explored for their applications in biomedical diagnostics, making their applications in healthcare biosensing a rapidly evolving field. Nanomaterials introduce versatility to the sensing platforms and may even allow mobility between different detection mechanisms. The prospect of a combination of different nanomaterials allows an exploitation of their synergistic additive and novel properties for sensor development. This paper covers more than 290 research works since 2015, elaborating the diverse roles played by various nanomaterials in the biosensing field. Hence, we provide a comprehensive review of the healthcare sensing applications of nanomaterials, covering carbon allotrope-based, inorganic, and organic nanomaterials. These sensing systems are able to detect a wide variety of clinically relevant molecules, like nucleic acids, viruses, bacteria, cancer antigens, pharmaceuticals and narcotic drugs, toxins, contaminants, as well as entire cells in various sensing media, ranging from buffers to more complex environments such as urine, blood or sputum. Thus, the latest advancements reviewed in this paper hold tremendous potential for the application of nanomaterials in the early screening of diseases and point-of-care testing.

/pdf/nanomaterials-for-healthcare-biosensing-applications-31xs4uxhju.pdf

Nanomaterials for Healthcare Biosensing Applications

Tumor exosomes are promising biomarkers for early cancer diagnosis in a noninvasive manner. However, precise capture and direct analysis of tumor-specific exosomes in complex biological samples are still challenging. Herein, we present a highly efficient dual-aptamer recognition system for precisely isolating and quantifying tumor exosomes from the complex biological environment based on hyperbranched DNA superstructure-facilitated signal amplification and ratiometric dual-signal strategies. When tumor exosomes were captured by the dual-aptamer recognition system, the cholesterol-modified DNA probe was anchored on the surface of the exosomes, activating DNA tetrahedron-based hyperbranched hybridization chain reaction to generate a sandwich complex. Then, the sandwich complex could bind a large number of Ru(NH3)63+ (Ru(III)), leading to a small amount of unbound Ru(III) left in the supernatant after magnetic separation. Hence, the redox reaction between Ru(II) and [Fe(CN)6]3- (Fe(III)) was significantly prevented, causing an obviously enhanced IFe(III)/IRu(III) value. Consequently, highly sensitive detection of tumor exosomes was achieved. The developed approach successfully realized direct isolation and analysis of tumor exosomes in complex sample media and human serum samples as well. More significantly, this ratiometric dual-signal mode and immobilization-free strategy effectively circumvented the systematic errors caused by external factors and the tedious probe immobilization processes, thus displaying the excellent performances of high reliability, improved accuracy, and easy manipulation. Overall, this approach is expected to offer novel ways for nondestructive early cancer diagnosis.

Precise Capture and Direct Quantification of Tumor Exosomes via a Highly Efficient Dual-Aptamer Recognition-Assisted Ratiometric Immobilization-Free Electrochemical Strategy.

Rapid and efficient tools for early cancer detection have diagnostic and therapeutic value. Given that the DNA hairpin‐based hybridization chain reaction (HCR) is effective in detecting various biological targets, a tetrahedral framework DNA‐enhanced (TDN‐enhanced) HCR detection system (T‐probe system) is introduced for cancer‐related targets and its versatility is demonstrated by detecting intracellular target miRNA 21 and cellular membrane target nucleolin. Benefiting from the spatial confinement of the TDN, the T‐probe system demonstrates a high detection rate. It increases the reaction efficiency of nude hairpins in vitro while accurately and rapidly recognizing both membrane and intracellular cancer‐related targets in living cells. Furthermore, it exhibits superior fluorescence in vivo within 15 s of peripheral‐tumor injection and 10 min of tail‐vein injection. The T‐probe system efficiently and accurately images tumor‐related biomarkers in vitro and in vivo, thereby demonstrating great potential for clinical diagnosis and therapeutic applications.

Facilitating In Situ Tumor Imaging with a Tetrahedral DNA Framework‐Enhanced Hybridization Chain Reaction Probe

A catalytic molecule machine-driven biosensing method for amplified electrochemical detection of exosomes.

Integration of fluorescence imaging and electrochemical biosensing for both qualitative location and quantitative detection of cancer cells.

Simple and universal signal labeling of cell surface for amplified detection of cancer cells via mild reduction

CD44, a stem-like biomarker, plays a critical role in maintenance of stem-like phenotype in breast cancer and is associated with migration, invasion and apoptosis resistance of breast tumors. Herein, we designed an effective sensing method using proximity-constructed bifunctional DNA probes for electrochemical identification of CD44. In our design, hybridization of capture probes was stabilized by target CD44 binding-induced proximity, thereby initiating subsequent cyclic strand displacement reactions. As a result, signaling probes were produced and dual-labeled with dibenzocyclooctyne group and CdTe quantum dots (QDs). These signaling probes performed two functions, specifically enrichment on azide-functionalized magnetic beads via click chemistry and generation of amplified electrochemical signaling relied on QDs. With aid of these bifunctional probes, the sensing method allowed not only highly specific identification of target CD44 even in serum samples and breast cancer cells but also monitoring of CD44 expression changes upon siRNA treatment. The linear range for CD44 identification was 0.1–1000 ng/mL and the detection limit was 0.0792 ng/mL, which displayed an improved sensitivity compared to previous reports. Therefore, the method may provide a powerful tool for CD44 identification and have a great potential in revealing stem-like properties in breast cancer in the future.

Proximity-constructed bifunctional DNA probes for identification of stem-like biomarker in breast cancer

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A catalytic molecule machine-driven biosensing method for amplified electrochemical detection of exosomes.

Integration of fluorescence imaging and electrochemical biosensing for both qualitative location and quantitative detection of cancer cells.

Simple and universal signal labeling of cell surface for amplified detection of cancer cells via mild reduction

Proximity-constructed bifunctional DNA probes for identification of stem-like biomarker in breast cancer