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.

Gold nanoparticle-based electrochemical biosensors

1.1. Significance of IVDs in the Clinic
IVD test is a crucial component of clinical care that performs a diagnostic test on biological samples that have been taken from a living body, such as blood, urine, and tissue.1 Such tests are usually performed to determine or confirm the presence of disease in an individual. “In vitro” literally means “within the glass”, which indicates that the test was historically conducted in glass test tubes. In contrast, in vivo tests, literally “within the living”, are conducted within a whole, living organism including human body.2 IVD tests have received much public attention because of their distinct features in the medical profession. First, IVD tests do not interact with the human body directly, making such diagnosis accessible without invasive surgeries and thus saving a great deal of suffering. Second, the procedures of IVDs are performed on samples rather than human body, avoiding the possible biological safety problems on patients that often take place in the in vivo diagnostics. Third, an IVD test can quickly provide valuable information on a patient’s healthcare conditions. On the basis of the information, physicians or patients are able to make a timely decision for patient care or treatment. Fourth, the application of IVDs enables early diagnosis and makes treatment of serious diseases easier. Generally, the cost of early testing is much lower than that of the later on extensive treatment. Last, IVDs play a particularly vital role in remote settings for managing outbreaks of acute infectious diseases, where effective but simple diagnostic systems are highly desirable. These features make IVDs unique and of great importance among medical technologies.

Gold Nanoparticles for In Vitro Diagnostics

Synthesis and electrochemical applications of gold nanoparticles

Recent developments in the field of screen-printed electrodes and their related applications.

A disposable electrochemical immunosensor for flow injection immunoassay of carcinoembryonic antigen

A gold nanoparticles/sol–gel composite architecture for encapsulation of immunoconjugate for reagentless electrochemical immunoassay

Background: Automated and convenient multianalyte detection with high throughput is increasingly needed in clinical diagnosis. We developed a disposable 4-by-2 array for programmed simultaneous amperometric immunoassay of 4 tumor markers. Methods: We used a screen-printed technique, 1-step immobilization method, and flow injection technique. We immobilized carcinoembryonic antigen, -fetoprotein, -human choriogonadotropin, and carcinoma antigen 125 as model analytes in a redox mediator–grafted, biopolymer-modified, screen-printed carbon electrode array to capture corresponding horseradish peroxidaselabeled antibodies in competitive immunoreactions. The simultaneous multianalyte immunoassay was automatically carried out to amperometrically monitor the mediator-catalyzed enzymatic response to hydrogen peroxide, which decreased in proportion to the concentrations of analytes in samples. Results: The multianalyte immunosensor array had a throughput of 60 samples/h and allowed simultaneous detection of carcinoembryonic antigen, -fetoprotein, -human choriogonadotropin, and carcinoma antigen 125 in clinical serum samples with concentrations up to 188 g/L, 250 g/L, 266 IU/L, and 334 kIU/L, respectively. The detection limits (limits of the blank, mean of blank plus 3 SD) were 1.1 g/L, 1.7 g/L, 1.2 IU/L, and 1.7 kIU/L. The inter- and intraassay imprecision (CVs) of the immunosensor arrays were <7.8% and <9.0%, respectively. The immunosensor arrays were stable for 28 days. Conclusions: This newly constructed immunosensor array provides a simple, automated, simultaneous multianalyte immunoassay with high throughput, short analytical time, and sufficiently low detection limits for clinical application. This method offers the capability of miniaturizing the multianalyte detection device. © 2007 American Association for Clinical Chemistry

A Disposable Multianalyte Electrochemical Immunosensor Array for Automated Simultaneous Determination of Tumor Markers

A two-dimensional resolution system of channels and substrate zones was proposed for multiplex immunoassay performed with a designed multichannel chemiluminescent (CL) detection device coupled with a single photomultiplier. Using carcinoma antigen 125 (CA 125), carcinoma antigen 153 (CA 153), carcinoma antigen 199 (CA 199), and carcinoembryonic antigen (CEA) as two couples of model analytes, two couples of capture antibodies were immobilized in two channels, respectively. With a sandwich format, the CL substrates for alkaline phosphatase and horseradish peroxidase were delivered into the channels sequentially to perform a multiplex immunoassay after the sample and tracer antibodies were introduced into the channels for on-line incubation. CA 125, CA 153, CA 199, and CEA could be assayed in the ranges of 0.50−80, 2.0−100, and 5.0−150 U/mL and 1.0−70 ng/mL with limits of detection of 0.15, 0.80, and 2.0 U/mL and 0.65 ng/mL at 3σ, respectively. The whole assay process including regeneration of the device cou...

Channel and Substrate Zone Two-Dimensional Resolution for Chemiluminescent Multiplex Immunoassay

Background: A reagentless sensor array for simultaneous multianalyte testing (SMAT) may enable accurate diagnosis and be applicable for point-of-care testing. We developed a disposable reagentless immunosensor array for simple immunoassay of panels of tumor markers.

Methods: We carried out SMAT with a direct capture format, in which colloidal gold nanoparticles with bound horseradish peroxidase (HRP)-labeled antibodies were immobilized on screen-printed carbon electrodes with biopolymer/sol-gel to trap their corresponding antigens from sample solution. Upon formation of immunocomplex, the direct electrochemical signal of the HRP decreased owing to increasing spatial blocking, and the analytes could be simultaneously determined by monitoring the signal changes.

Results: The proposed reagentless immunosensor array allowed simultaneous detection of carcinoma antigen 153, carcinoma antigen 125, carbohydrate antigen 199, and carcinoembryonic antigen in clinical serum samples in the ranges of 0.4–140 kU/L, 0.5–330 kU/L, 0.8–190 kU/L, and 0.1–44 μg/L, respectively, with detection limits of 0.2 kU/L, 0.5 kU/L, 0.3 kU/L, and 0.1 μg/L corresponding to the signals 3 SD above the mean of a zero standard. The interassay imprecision of the arrays was 95.5% for 95 cases of cancer-positive sera.

Conclusions: The immunosensor array provides a SMAT with short analytical time, small sampling volume, no need for substrate, and, no between-electrode cross-talk. This method not only proved the capability of the array in point-of-care testing, but also allowed simultaneous testing of several tumor markers.

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Jinhai Tang

Papers

A disposable electrochemical immunosensor for flow injection immunoassay of carcinoembryonic antigen

A gold nanoparticles/sol–gel composite architecture for encapsulation of immunoconjugate for reagentless electrochemical immunoassay

A Disposable Multianalyte Electrochemical Immunosensor Array for Automated Simultaneous Determination of Tumor Markers

Channel and Substrate Zone Two-Dimensional Resolution for Chemiluminescent Multiplex Immunoassay

Disposable Reagentless Electrochemical Immunosensor Array Based on a Biopolymer/Sol-Gel Membrane for Simultaneous Measurement of Several Tumor Markers