Detection of chemical and biological agents plays a fundamental role in biomedical, forensic and environmental sciences1–4 as well as in anti bioterrorism applications.5–7 The development of highly sensitive, cost effective, miniature sensors is therefore in high demand which requires advanced technology coupled with fundamental knowledge in chemistry, biology and material sciences.8–13

In general, sensors feature two functional components: a recognition element to provide selective/specific binding with the target analytes and a transducer component for signaling the binding event. An efficient sensor relies heavily on these two essential components for the recognition process in terms of response time, signal to noise (S/N) ratio, selectivity and limits of detection (LOD).14,15 Therefore, designing sensors with higher efficacy depends on the development of novel materials to improve both the recognition and transduction processes. Nanomaterials feature unique physicochemical properties that can be of great utility in creating new recognition and transduction processes for chemical and biological sensors15–27 as well as improving the S/N ratio by miniaturization of the sensor elements.28

Gold nanoparticles (AuNPs) possess distinct physical and chemical attributes that make them excellent scaffolds for the fabrication of novel chemical and biological sensors (Figure 1).29–36 First, AuNPs can be synthesized in a straightforward manner and can be made highly stable. Second, they possess unique optoelectronic properties. Third, they provide high surface-to-volume ratio with excellent biocompatibility using appropriate ligands.30 Fourth, these properties of AuNPs can be readily tuned varying their size, shape and the surrounding chemical environment. For example, the binding event between recognition element and the analyte can alter physicochemical properties of transducer AuNPs, such as plasmon resonance absorption, conductivity, redox behavior, etc. that in turn can generate a detectable response signal. Finally, AuNPs offer a suitable platform for multi-functionalization with a wide range of organic or biological ligands for the selective binding and detection of small molecules and biological targets.30–32,36 Each of these attributes of AuNPs has allowed researchers to develop novel sensing strategies with improved sensitivity, stability and selectivity. In the last decade of research, the advent of AuNP as a sensory element provided us a broad spectrum of innovative approaches for the detection of metal ions, small molecules, proteins, nucleic acids, malignant cells, etc. in a rapid and efficient manner.37



Figure 1

Physical properties of AuNPs and schematic illustration of an AuNP-based detection system.



In this current review, we have highlighted the several synthetic routes and properties of AuNPs that make them excellent probes for different sensing strategies. Furthermore, we will discuss various sensing strategies and major advances in the last two decades of research utilizing AuNPs in the detection of variety of target analytes including metal ions, organic molecules, proteins, nucleic acids, and microorganisms.

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Gold nanoparticles in chemical and biological sensing.

Polydopamine and Its Derivative Materials: Synthesis and Promising Applications in Energy, Environmental, and Biomedical Fields

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Principles Of Fluorescence Spectroscopy

Chemistries that Facilitate Nanotechnology Kim E. Sapsford,† W. Russ Algar, Lorenzo Berti, Kelly Boeneman Gemmill,‡ Brendan J. Casey,† Eunkeu Oh, Michael H. Stewart, and Igor L. Medintz*,‡ †Division of Biology, Department of Chemistry and Materials Science, Office of Science and Engineering Laboratories, U.S. Food and Drug Administration, Silver Spring, Maryland 20993, United States ‡Center for Bio/Molecular Science and Engineering Code 6900 and Division of Optical Sciences Code 5611, U.S. Naval Research Laboratory, Washington, D.C. 20375, United States College of Science, George Mason University, 4400 University Drive, Fairfax, Virginia 22030, United States Department of Biochemistry and Molecular Medicine, University of California, Davis, School of Medicine, Sacramento, California 95817, United States Sotera Defense Solutions, Crofton, Maryland 21114, United States

Functionalizing nanoparticles with biological molecules: developing chemistries that facilitate nanotechnology.

Isothermal amplification of nucleic acids is a simple process that rapidly and efficiently accumulates nucleic acid sequences at constant temperature. Since the early 1990s, various isothermal amplification techniques have been developed as alternatives to polymerase chain reaction (PCR). These isothermal amplification methods have been used for biosensing targets such as DNA, RNA, cells, proteins, small molecules, and ions. The applications of these techniques for in situ or intracellular bioimaging and sequencing have been amply demonstrated. Amplicons produced by isothermal amplification methods have also been utilized to construct versatile nucleic acid nanomaterials for promising applications in biomedicine, bioimaging, and biosensing. The integration of isothermal amplification into microsystems or portable devices improves nucleic acid-based on-site assays and confers high sensitivity. Single-cell and single-molecule analyses have also been implemented based on integrated microfluidic systems. In this review, we provide a comprehensive overview of the isothermal amplification of nucleic acids encompassing work published in the past two decades. First, different isothermal amplification techniques are classified into three types based on reaction kinetics. Then, we summarize the applications of isothermal amplification in bioanalysis, diagnostics, nanotechnology, materials science, and device integration. Finally, several challenges and perspectives in the field are discussed.

Isothermal Amplification of Nucleic Acids

DNA methylation catalyzed by methyltransferase (MTase) is a significant epigenetic process for modulating gene expression. Traditional methods to study MTase activity require a laborious and costly DNA labeling process. In this article, we report a simple, colorimetric, and label-free methylation-responsive DNAzyme (MR-DNAzyme) strategy for MTase activity analysis. This new strategy relies on horseradish peroxidase (HRP) mimicking DNAzyme and the methylation-responsive sequence (MRS) of DNA which can be methylated and cleaved by the MTase/endonuclease coupling reaction. Methylation-induced scission of MRS would activate the DNAzyme that can catalyze the generation of a color signal for the amplified detection of methylation events. Taking Dam MTase and DpnI endonuclease as examples, we have developed two colorimetric methods based on the MR-DNAzyme strategy. The first method is to utilize an engineered hairpin-DNAzyme hybrid probe for facile turn-on detection of Dam MTase activity, with a wide linear rang...

Label-free colorimetric assay for methyltransferase activity based on a novel methylation-responsive DNAzyme strategy.

New polymer–enzyme–metallic nanoparticle composite films with a high-load and a high-activity of immobilized enzymes and obvious electrocatalysis/nano-enhancement effects for biosensing of glucose and galactose are designed and prepared by a one-pot chemical pre-synthesis/electropolymerization (CPSE) protocol. Dopamine (DA) as a reductant and a monomer, glucose oxidase (GOx) or galactose oxidase (GaOx) as the enzyme, and HAuCl4 or H2PtCl6 as an oxidant to trigger DA polymerization and the source of metallic nanoparticles, are mixed to yield polymeric bionanocomposites (PBNCs), which are then anchored on the electrode by electropolymerization of the remaining DA monomer. The prepared PBNC material has good biocompatibility, a highly uniform dispersion of the nanoparticles with a narrow size distribution, and high load/activity of the immobilized enzymes, as verified by transmission/scanning electron microscopy and electrochemical quartz crystal microbalance. The thus-prepared enzyme electrodes show a largely improved amperometric biosensing performance, e.g., a very high detection sensitivity (99 or 129 µA cm−2 mM−1 for glucose for Pt PBNCs on bare or platinized Au), a sub-micromolar limit of detection for glucose, and an excellent durability, in comparison with those based on conventional procedures. Also, the PBNC-based enzyme electrodes work well in the second-generation biosensing mode. The proposed one-pot CPSE protocol may be extended to the preparation of many other functionalized PBNCs for wide applications.

One-Pot Preparation of Polymer–Enzyme–Metallic Nanoparticle Composite Films for High-Performance Biosensing of Glucose and Galactose

Herein, we present a novel label-free fluorescent assay for monitoring the activity and inhibition of protein kinases based on the aggregation behavior of unmodified CdTe quantum dots (QDs). In this assay, cationic substrate peptides induce the selective aggregation of unmodified QDs with anionic surface charge, whereas phosphorylated peptides do not. Phosphorylation by kinase alters the net charge of peptides and subsequently inhibits the aggregation of unmodified QDs, causing an enhanced fluorescence with a 45 nm blue-shift in emission and a yellow-to-green emission color change. Hence the fluorescence response allows this QD-based method to easily probe kinase activity by a spectrometer or even by the naked eye. The feasibility of the method has been demonstrated by sensitive measurement of the activity of cAMP-dependent protein kinase (PKA) with a low detection limit (0.47 mU μL−1). On the basis of the fluorescence response of QDs on the concentration of PKA inhibitor H-89, the IC50 value, the half ma...

Label-Free Fluorescent Detection of Protein Kinase Activity Based on the Aggregation Behavior of Unmodified Quantum Dots

Metal-organic frameworks (MOFs) have shown considerable prospects for sensing pesticide residues. However, the low stability of MOFs in water hinders them from testing food and environmental samples. Herein, we report an easy and cost-efficient synthesis of a water-stable zirconium luminescent MOF (Zr-LMOF) and its application for rapid, sensitive, and in situ detection of organophosphorous pesticides (OPPs). The Zr-MOF is prepared using Zr(IV) and 1,2,4,5-tetrakis(4-carboxyphenyl)benzene. The synthesized Zr-LMOF rapidly absorbs trace amounts of OPP parathion-methyl and indicates its presence. A low limit of detection of 0.115 μg kg-1 (0.438 nM) with a wide linear range from 70 μg kg-1 to 5.0 mg kg-1 was achieved. Satisfactory recoveries ranging from 78% to 107% were obtained for spiked food and environmental samples. Further, the Zr-LMOF was applied to imitate rapid and in situ imaging detection of pesticide residue on fresh produce nondestructively; visual signals appeared under ultraviolet light within 5 min. These results suggest that the Zr-LMOF has the potential for low-cost, rapid, and in situ imaging detection of OPPs contamination via easy-to-read visual signal.

A Water-Stable Luminescent Metal-Organic Framework for Rapid and Visible Sensing of Organophosphorus Pesticides.

This work reports a highly luminescent metal-organic framework (MOF) with high surface area of 4073.9 m2 g−1, exhibiting an excellent adsorption ability and sensing performance in rapidly detecting parathion-methyl. This is an excellent example of luminescent MOFs to sensitively detect pesticides by fluorescent quenching with satisfactory results. This sensor provides a very wide linear detection range from 1.0 μg kg−1 to 10 mg kg−1, and low limit of detection 0.12 μg kg−1 (0.456 nM) for parathion-methyl. This low detection limit is much lower than the maximum residue limits (MRL) in the European Union pesticides database (0.01 mg kg−1 for fruits and vegetables, 0.02 mg kg−1 for tree nuts), as well as those of previous assays. The quenching effect of parathion-methyl is attributed to the electron-rich chemical group of nitroaromatics which might lead to photo-induced electron transfer. The fluorescent method has been reliably used for the determination of parathion-methyl in spiked irrigation water with satisfactory results, which suggests its potential for rapid detection of pesticides in food and environmental sample.

Xiahong Xu

Papers

Label-free colorimetric assay for methyltransferase activity based on a novel methylation-responsive DNAzyme strategy.

One-Pot Preparation of Polymer–Enzyme–Metallic Nanoparticle Composite Films for High-Performance Biosensing of Glucose and Galactose

Label-Free Fluorescent Detection of Protein Kinase Activity Based on the Aggregation Behavior of Unmodified Quantum Dots

A Water-Stable Luminescent Metal-Organic Framework for Rapid and Visible Sensing of Organophosphorus Pesticides.

Sensitive detection of pesticides by a highly luminescent metal-organic framework