Owing to their excellent multiplexing ability, high sensitivity, and large dynamic range, immunoassays using surface-enhanced Raman scattering (SERS) as the readout signal have found prosperous applications in fields such as disease diagnosis, environmental surveillance, and food safety supervision. Various ever-increasing demands have promoted SERS-based immunoassays from the classical sandwich-type ones to those integrated with fascinating automatic platforms (e.g., test strips and microfluidic chips). As recent years have witnessed impressive progress in SERS immunoassays, we try to comprehensively cover SERS-based immunoassays from their basic working principles to specific applications. Focusing on several basic elements in SERS immunoassays, typical structures of SERS nanoprobes, productive optical spectral encoding strategies, and popular immunoassay platforms are highlighted, followed by their representative biological applications in the last 5 years. Moreover, despite the vast advances achieved ...

SERS-Activated Platforms for Immunoassay: Probes, Encoding Methods, and Applications

In this work, a sensitive and selective ratiometric fluorescence sensing platform was built for the detection of tyrosinase (TYR) activity and dopamine (DA) using glutathione (GSH) protected gold nanoclusters (Au NCs) as probes. Upon excitation at 350 nm, Au NCs displayed an intense red emission, which could be effectively quenched by quinones. TYR, a typical polyphenol oxidase, can catalyze the oxidization of DA to o-quinone and therefore quenched the fluorescence of Au NCs. Moreover, the reaction of TYR and DA gave rise to an emission band at 400 nm, which increased in a TYR/DA-concentration-dependent manner. The ratiometric signal variations were utilized for facile, sensitive, and selective detection of TYR activity and DA. A linear range was obtained from 0.006–3.6 unit mL–1 of TYR activity, while the linear range for detection of DA was 1.0 nM to 1.0 mM. Additionally, it constructed a useful platform for TYR inhibitor screening in biomedical research.

Ratiometric Fluorescence Detection of Tyrosinase Activity and Dopamine Using Thiolate-Protected Gold Nanoclusters

Neurotransmitters are chemicals that act as messengers in the synaptic transmission process. They are essential for human health and any imbalance in their activities can cause serious mental disorders such as Parkinson’s disease, schizophrenia, and Alzheimer’s disease. Hence, monitoring the concentrations of various neurotransmitters is of great importance in studying and diagnosing such mental illnesses. Recently, many researchers have explored the use of unique materials for developing biosensors for both in vivo and ex vivo neurotransmitter detection. A combination of nanomaterials, polymers, and biomolecules were incorporated to implement such sensor devices. For in vivo detection, electrochemical sensing has been commonly applied, with fast-scan cyclic voltammetry being the most promising technique to date, due to the advantages such as easy miniaturization, simple device architecture, and high sensitivity. However, the main challenges for in vivo electrochemical neurotransmitter sensors are limited target selectivity, large background signal and noise, and device fouling and degradation over time. Therefore, achieving simultaneous detection of multiple neurotransmitters in real time with long-term stability remains the focus of research. The purpose of this review paper is to summarize the recently developed sensing techniques with the focus on neurotransmitters as the target analyte, and to discuss the outlook of simultaneous detection of multiple neurotransmitter species. This paper is organized as follows: firstly, the common materials used for developing neurotransmitter sensors are discussed. Secondly, several sensor surface modification approaches to enhance sensing performance are reviewed. Finally, we discuss recent developments in the simultaneous detection capability of multiple neurotransmitters.

Recent Advances in the Detection of Neurotransmitters

Facile green synthesis of stable silver nanoparticles in flower extract of Acmella oleracea and its dopamine sensing properties are reported. UV–visible spectroscopy, Fourier transform infrared (FTIR) spectroscopy, transmission electron microscopy (TEM) and particle size analysis were performed to ascertain the formation of silver nanoparticles. Aqueous dopamine sensing study of green synthesized silver nanoparticles was carried out. Further surface plasmon resonance (SPR) based fiber optic sensor has been fabricated using green synthesized silver nanoparticles as sensing material. The response time of the sensor is 6 min and the detection limit is 2 × 10−7 M.

Surface plasmon resonance based fiber optic dopamine sensor using green synthesized silver nanoparticles

Electrochemical detection of carcinoembryonic antigen

A new mercapto-terminated binuclear Cu(II) complex, [Cu2Cl2(mtboc)2(C12H8N2)2] [mtboc =2-(methylthio)benzo[d]oxazole-5-carboxylic acid] (Cu2), was synthesized and characterized by single-crystal X-ray diffraction studies. The Cu2 complex was then immobilized on the surface of Au electrodes through S Au bonds to form a self-assembled monolayer. The electrochemical properties of the as-prepared Cu2- modified Au electrode (Au/Cu2) were investigated using cyclic voltammetry, electrochemical impedance spectroscopy, and differential pulse voltammetry (DPV). Moreover, as a biomimetic sensor for the determination of dopamine (DA) and uric acid (UA), the Au/Cu2 electrode exhibited excellent electrocatalytic activity for the oxidation of DA with a faster electron-transfer rate (6.61 s−1). The DPV technique was used for the trace determination of DA. The dependence of current vs. concentration was linear from 0.2 to 30 μM with a regression coefficient of 0.9975, and the detection limit of DA was ∼ 0.08 μM. Furthermore, the modified electrode clearly separated the DA signal from the interference effect of UA, thus realizing the simultaneous detection of DA and UA. Finally, the method was applied to the selective and precise analysis of DA in commercial injection and urine samples.

Application of a mercapto-terminated binuclear Cu(II) complex modified Au electrode to improve the sensitivity and selectivity for dopamine detection

A simple and sensitive method, based on surface-enhanced Raman scattering (SERS), for immunoassay and label-free protein detection is reported. A series of bowl-shaped silver cavity arrays were fabricated by electrodeposition using a self-assembled polystyrene spheres template. The reflection spectra of these cavity arrays were recorded as a function of film thickness, and then correlated with SERS enhancement using sodium thiophenolate as the probe molecule. The results reveal that SERS enhancement can be maximized when the frequency of both the incident laser and the Raman scattering approach the frequency of the localized surface plasmon resonance. The optimized array was then used as the bottom layer of a silver nanoparticle–protein–bowl-shaped silver cavity array sandwich. The second layer of silver was introduced by the interactions between the proteins in the middle layer of the sandwich architecture and silver nanoparticles. Human IgG bound to the surface of this microcavity array can retain its recognition function. With the Raman reporter molecules labeled on the antibody, a detection limit down to 0.1 ng mL−1 for human IgG is easily achieved. Furthermore, the SERS spectra of label-free proteins (catalase, cytochrome C, avidin and lysozyme) from the assembled sandwich have excellent reproducibility and high quality. The results reveal that the proposed approach has potential for use in qualitative and quantitative detection of biomolecules.

Using a silver-enhanced microarray sandwich structure to improve SERS sensitivity for protein detection

Detection of dopamine on a mercapto-terminated hexanuclear Fe(III) cluster modified gold electrode.

A surface-enhanced Raman scattering (SERS)-based competitive immunoassay for detecting β-adrenergic agonists (salbutamol, SAL; brombuterol, BRO) was developed. Highly ordered gold cavity arrays (GCA) with adjustable localized surface plasmon resonance and great Raman enhancement were prepared by electrodepositing gold nanoparticles (NPS) to the interstices of polystyrene sphere templates, and then covered by a pre-punched sticker with many holes as detection areas in the assay. The immuno-probe was prepared by immobilizing antibody against SAL (or BRO) and DNA concatamers labelled with SERS reporter on gold NPS. After immunoreaction, the characteristic bands of SERS reporters captured on GCA were measured for quantification of SAL and BRO. The sensitivity was improved by introduction of long DNA concatamers and silver staining due to DNA amplification and strong plasmon coupling between Au and Ag NPS. The LODs for SAL and BRO were 2.0 pg mL−1 and 1.0 pg mL−1, respectively. The recoveries of the assay for SAL and BRO in spiked samples were 96.7∼105.3 % with RSD of 4.9–8.7 %. The results demonstrated that this approach was a sensitive, specific and high-throughput method for simultaneous detection of SAL and BRO, and might be a new platform for the detection of multiple compounds in food and biological fields.

A SERS-based competitive immunoassay using highly ordered gold cavity arrays as the substrate for simultaneous detection of β-adrenergic agonists

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Application of a mercapto-terminated binuclear Cu(II) complex modified Au electrode to improve the sensitivity and selectivity for dopamine detection

Using a silver-enhanced microarray sandwich structure to improve SERS sensitivity for protein detection

Detection of dopamine on a mercapto-terminated hexanuclear Fe(III) cluster modified gold electrode.

A SERS-based competitive immunoassay using highly ordered gold cavity arrays as the substrate for simultaneous detection of β-adrenergic agonists