Lateral flow immunoassay (LFIA) is a widely used express method and offers advantages such as a short analysis time, simplicity of testing and result evaluation. However, an LFIA based on gold nanospheres lacks the desired sensitivity, thereby limiting its wide applications. In this study, spherical nanogold labels along with new types of nanogold labels such as gold nanopopcorns and nanostars were prepared, characterized, and applied for LFIA of model protein antigen procalcitonin. It was found that the label with a structure close to spherical provided more uniform distribution of specific antibodies on its surface, indicative of its suitability for this type of analysis. LFIA using gold nanopopcorns as a label allowed procalcitonin detection over a linear range of 0.5–10 ng mL−1 with the limit of detection of 0.1 ng mL−1, which was fivefold higher than the sensitivity of the assay with gold nanospheres. Another approach to improve the sensitivity of the assay included the silver enhancement method, which was used to compare the amplification of LFIA for procalcitonin detection. The sensitivity of procalcitonin determination by this method was 10 times better the sensitivity of the conventional LFIA with gold nanosphere as a label. The proposed approach of LFIA based on gold nanopopcorns improved the detection sensitivity without additional steps and prevented the increased consumption of specific reagents (antibodies).
The growing interest in the development of new platforms for the application of Raman spectroscopy techniques in biosensor technologies is driven by the potential of these techniques in identifying chemical compounds, as well as structural and functional features of biomolecules. The effect of Raman scattering is a result of inelastic light scattering processes, which lead to the emission of scattered light with a different frequency associated with molecular vibrations of the identified molecule. Spontaneous Raman scattering is usually weak, resulting in complexities with the separation of weak inelastically scattered light and intense Rayleigh scattering. These limitations have led to the development of various techniques for enhancing Raman scattering, including resonance Raman spectroscopy (RRS) and nonlinear Raman spectroscopy (coherent anti-Stokes Raman spectroscopy and stimulated Raman spectroscopy). Furthermore, the discovery of the phenomenon of enhanced Raman scattering near metallic nanostructures gave impetus to the development of the surface-enhanced Raman spectroscopy (SERS) as well as its combination with resonance Raman spectroscopy and nonlinear Raman spectroscopic techniques. The combination of nonlinear and resonant optical effects with metal substrates or nanoparticles can be used to increase speed, spatial resolution, and signal amplification in Raman spectroscopy, making these techniques promising for the analysis and characterization of biological samples. This review provides the main provisions of the listed Raman techniques and the advantages and limitations present when applied to life sciences research. The recent advances in SERS and SERS-combined techniques are summarized, such as SERRS, SE-CARS, and SE-SRS for bioimaging and the biosensing of molecules, which form the basis for potential future applications of these techniques in biosensor technology. In addition, an overview is given of the main tools for success in the development of biosensors based on Raman spectroscopy techniques, which can be achieved by choosing one or a combination of the following approaches: (i) fabrication of a reproducible SERS substrate, (ii) synthesis of the SERS nanotag, and (iii) implementation of new platforms for on-site testing.
The current COVID-19 pandemic has increased the demand for pathogen detection methods that combine low detection limits with rapid results. Despite the significant progress in methods and devices for nucleic acid amplification, immunochemical methods are still preferred for mass testing without specialized laboratories and highly qualified personnel. The most widely used immunoassays are microplate enzyme-linked immunosorbent assay (ELISA) with photometric detection and lateral flow immunoassay (LFIA) with visual results assessment. However, the disadvantage of ELISA is its considerable duration, and that of LFIA is its low sensitivity. In this study, the modified LFIA of a specific antigen of the causative agent of COVID-19, spike receptor-binding domain, was developed and characterized. This modified LFIA includes the use of gold nanoparticles with immobilized antibodies and 4-mercaptobenzoic acid as surface-enhanced Raman scattering (SERS) nanotag and registration of the nanotag binding by SERS spectrometry. To enhance the sensitivity of LFIA-SERS analysis, we determined the optimal compositions of SERS nanotags and membranes used in LFIA. For benchmark comparison, ELISA and conventional colorimetric LFIA were used with the same immune reagents. The proposed method combines a low detection limit of 0.1 ng/mL (at 0.4 ng/mL for ELISA and 1 ng/mL for qualitative LFIA) with a short assay time equal to 20 min (at 3.5 h for ELISA and 15 min for LFIA). The results obtained demonstrate the promise of using the SERS effects in membrane immuno-analytical systems.
In this study, new types of nanogold labels for lateral flow immunoassay of model antigen procalcitonin based on photometric and surface-enhanced Raman scattering methods of detection were obtained. The linear range of procalcitonin determination applying gold nanoflowers as a label was between 0.5 and 10 ng/mL. The limit of photometric detection was achieved at 0.1 ng/mL that was five times lower than the sensitivity of traditional lateral flow immunoassay with spherical gold nanoparticles as a label. In addition, the conjugate of polyclonal antibodies against procalcitonin with spherical gold nanoparticles labeled with 4-mercaptobenzoic acid as Raman reporter molecule was prepared and used as an immunoprobe in lateral flow immunoassay based on surface-enhanced Raman spectroscopy to improve assay sensitivity.
In this study, tannic acid-modified gold nanoparticles were found to have superior nanozyme activity and catalyze the oxidation reaction of 3,3′,5,5′-tetramethylbenzidine in the presence of hydrogen peroxide. Enhancing the catalytic activity of the nanozyme by Pb2+ ions caused by selectively binding metal ions by the tannic acid-capped surface of gold nanoparticles makes them an ideal colorimetric probe for Pb2+. The parameters of the reaction, including pH, incubation time, and concentration of components, were optimized to reach maximal sensitivity of Pb2+ detection. The absorption change is directly proportional to the Pb2+ concentration and allows the determination of Pb2+ ions within 10 min. The colorimetric sensor is characterized by a wide linear range from 25 to 500 ng×mL−1 with a low limit of detection of 11.3 ng×mL−1. The highly sensitive and selective Pb2+ detection in tap, drinking, and spring water revealed the feasibility and applicability of the developed colorimetric sensor.
The development of reliable and highly sensitive methods for heavy metal detection is a critical task for protecting the environment and human health. In this study, a qualitative colorimetric sensor that used mercaptosuccinic-acid-functionalized gold nanoparticles (MSA-AuNPs) to detect trace amounts of Fe(III) ions was developed. MSA-AuNPs were prepared using a one-step reaction, where mercaptosuccinic acid (MSA) was used for both stabilization, which was provided by the presence of two carboxyl groups, and functionalization of the gold nanoparticle (AuNP) surface. The chelating properties of MSA in the presence of Fe(III) ions and the concentration-dependent aggregation of AuNPs showed the effectiveness of MSA-AuNPs as a sensing probe with the use of an absorbance ratio of A530/A650 as an analytical signal in the developed qualitative assay. Furthermore, the obvious Fe(III)-dependent change in the color of the MSA-AuNP solution from red to gray-blue made it possible to visually assess the metal content in a concentration above the detection limit with an assay time of less than 1 min. The detection limit that was achieved (23 ng/mL) using the proposed colorimetric sensor is more than 10 times lower than the maximum allowable concentration for drinking water defined by the World Health Organization (WHO). The MSA-AuNPs were successfully applied for Fe(III) determination in tap, spring, and drinking water, with a recovery range from 89.6 to 126%. Thus, the practicality of the MSA-AuNP-based sensor and its potential for detecting Fe(III) in real water samples were confirmed by the rapidity of testing and its high sensitivity and selectivity in the presence of competing metal ions.
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