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1.
Anal Chem ; 2023 Jan 06.
Article in English | MEDLINE | ID: mdl-36608282

ABSTRACT

Point-of-care (POC) diagnostics in particular focuses on the timely identification of harmful conditions close to the patients' needs. For future healthcare these diagnostics could be an invaluable tool especially in a digitalized or telemedicine-based system. However, while paper-based POC tests, with the most prominent example being the lateral flow assay (LFA), have been especially successful due to their simplicity and timely response, the COVID-19 pandemic highlighted their limitations, such as low sensitivity and ambiguous responses. This perspective discusses strategies that are currently being pursued to evolve such paper-based POC tests toward a superior diagnostic tool that provides high sensitivities, objective result interpretation, and multiplexing options. Here, we pinpoint the challenges with respect to (i) measurability and (ii) public applicability, exemplified with select cases. Furthermore, we highlight promising endeavors focused on (iii) increasing the sensitivity, (iv) multiplexing capability, and (v) objective evaluation to also ready the technology for integration with machine learning into digital diagnostics and telemedicine. The status quo in academic research and industry is outlined, and the likely highly relevant role of paper-based POC tests in future healthcare is suggested.

2.
Mikrochim Acta ; 189(3): 117, 2022 02 23.
Article in English | MEDLINE | ID: mdl-35195801

ABSTRACT

Gold nanoparticle-catalyzed chemiluminescence (CL) of luminol is an attractive alternative to strategies relying on enzymes, as their aggregation leads to significantly enhanced CL signals. Consequently, analytes disturbing such aggregation will lead to an easy-to-quantify weakening of the signal. Based on this concept, a homogeneous aptamer-based assay for the detection of sulfadimethoxine (SDM) has been developed as a microfluidic CL flow-injection platform. Here, the efficient mixing of gold nanoparticles, aptamers, and analyte in short channel distances is of utmost importance, and two-dimensional (2D) and three-dimensional (3D) mixer designs made via Xurography were investigated. In the end, since 2D designs could not provide sufficient mixing, a laminated 3D 5-layer microfluidic mixer was developed and optimized with respect to mixing capability and observation by the charge-coupled device (CCD) camera. Furthermore, the performance of standard luminol and its more hydrophilic derivative m-carboxy luminol was studied identifying the hydrophilic derivative to provide tenfold more signal enhancement and reliable results. Finally, the novel detection platform was used for the specific detection of SDM via its aptamer and yielded a stunning dynamic range over 5 orders of magnitude (0.01-1000 ng/ml) and a limit of detection of 4 pg/ml. This new detection concept not only outperforms other methods for SDM detection, but can be suggested as a new flow-injection strategy for aptamer-based rapid and cost-efficient analysis in environmental monitoring and food safety.


Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , Metal Nanoparticles , Biosensing Techniques/methods , Gold , Luminescence , Luminescent Measurements/methods , Microfluidics , Sulfadimethoxine
3.
Anal Bioanal Chem ; 414(10): 3231-3241, 2022 Apr.
Article in English | MEDLINE | ID: mdl-34773470

ABSTRACT

Recent years have confirmed the ubiquitous applicability of lateral flow assays (LFA) in point-of-care testing (POCT). To make this technology available for low abundance analytes, strategies towards lower limits of detections (LOD), while maintaining the LFA's ease of use, are still being sought. Here, we demonstrate how liposomes can significantly improve the LOD of traditional gold nanoparticle (AuNP)-based assays while fully supporting a ready-to-use system for commercial application. We fine-tuned liposomes towards photometric and fluorescence performance on the synthesis level and applied them in an established interleukin 6 (IL-6) immunoassay normally using commercial AuNP labels. IL-6's low abundance (< 10 pg mL-1) and increasing relevance as prognostic marker for infections make it an ideal model analyte. It was found that liposomes with a high encapsulant load (150 mmol L-1 sulforhodamine B (SRB)) easily outperform AuNPs in photometric LFAs. Specifically, liposomes with 350 nm in diameter yield a lower LOD even in complex matrices such as human serum below the clinically relevant range (7 pg mL-1) beating AuNP by over an order of magnitude (81 pg mL-1). When dehydrated on the strip, liposomes maintained their signal performance for over a year even when stored at ambient temperature and indicate extraordinary stability of up to 8 years when stored as liquid. Whereas no LOD improvement was obtained by exploiting the liposomes' fluorescence, an extraordinary gain in signal intensity was achieved upon lysis which is a promising feature for high-resolution and low-cost detection devices. Minimizing the procedural steps by inherently fluorescent liposomes, however, is not feasible. Finally, liposomes are ready for commercial applications as they are easy to mass-produce and can simply be substituted for the ubiquitously used AuNPs in the POCT market.


Subject(s)
Gold , Metal Nanoparticles , Humans , Immunoassay , Interleukin-6 , Liposomes
4.
Anal Chim Acta ; 1188: 339161, 2021 Dec 15.
Article in English | MEDLINE | ID: mdl-34794566

ABSTRACT

Chemiluminescence (CL) provides outstanding analytical performance due to its independence from external light sources, background-free nature and exceptional sensitivity and selectivity. Yet, ultra-sensitive (bio)analysis is impeded by low hydrophilicity, poor quantum yields, fast kinetics or instability of most CL reagents such as luminol, acridinium esters, dioxetanes or peroxyoxalic derivatives. Photophysical studies show that m-carboxy luminol overcomes these limitations as its hydrophilic design provides a 5-fold increase in relative quantum yield resulting in superior performance in H2O2-dependent bioassays with 18-fold higher sensitivity for the quantification of its co-reactant H2O2, and 5-times lower detection limits for the luminophore. Studies with CL enhancers suggest its significance for mechanistic investigations in tandem with peroxidases. Finally, its integration into enzymatic and immunoassay applications demonstrates that m-carboxy luminol will provide signal enhancement, lower detection limits, and increased dynamic ranges for any other luminol-based CL assay, thus comprising the potential to replace luminol as benchmark probe.


Subject(s)
Luminescence , Luminol , Benchmarking , Hydrogen Peroxide , Luminescent Measurements
5.
Biosens Bioelectron ; 164: 112272, 2020 Sep 15.
Article in English | MEDLINE | ID: mdl-32553348

ABSTRACT

Highly porous laser-induced graphene (LIG) is easily generated in complex electrode configurations such as interdigitated electrodes (IDEs). Here, we demonstrate that their superior capacitive response at low frequencies can be exploited in affinity biosensors using thrombin aptamers as model biorecognition elements. Of specific interest was the effect of electrode surface area on capacitance detection, and the comparison between a label-free format and enhancement strategies afforded by carboxy group bearing polymeric nanoparticles or liposomes. Electrochemical impedance spectroscopy (EIS) was used to investigate the LIG performance and optimize the biosensor design. Interestingly, the label-free strategy performed extremely well and additional labels decreased the limit of detection or increased the sensitivity only minimally. It is assumed that the highly porous nature of the LIG structures dominates the capacitive response so that labels removed from the surface have only limited influence Also, while slight performance changes can be observed for smaller vs. larger electrode structures, the performance of a LIG IDE is reasonably independent of its size. In the end, a dynamic range of 5 orders of magnitude was obtained (0.01 nM-1000 nM) with a limit of detection as low as 0.12 pM. When measured in serum, this increased to 1.3 pM. The good reproducibility (relative standard deviation (RSD), 4.90%) and repeatability (RSD, 2.59%) and good long-term stability (>7 weeks at 4 °C) prove that a LIG-based capacitance sensor is an excellent choice for affinity-based biosensor. The ease-of-production, the simplicity of modification and the superior performance even in a label-free format indicate that LIG-based biosensors should be considered in point-of-care diagnostics in the future.


Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , Graphite , Electrodes , Lasers , Reproducibility of Results
6.
Anal Chem ; 91(20): 13080-13087, 2019 10 15.
Article in English | MEDLINE | ID: mdl-31524378

ABSTRACT

Luminol is a major probe for chemiluminescence (CL) and electrochemiluminescence (ECL) detection technologies in (bio)analysis. Surfactants are added to ECL assay cocktails to enhance signals or are present, owing to given bioassay protocols, yet little is known regarding their effects on luminol ECL. In-depth understanding is provided here through a broad study with bioanalytically relevant surfactants (cationic, anionic, and nonionic), four common electrode materials, and two luminol derivatives. Naturally, in ECL, surface effects are dominant; however, bulk solution, diffusion, and luminescence-stabilization processes also contribute significantly to the overall reaction. It was found that in contrast to CL the effect surfactants have on luminol ECL cannot be linked to general surfactant characteristics such as ionic nature, hydrophilic lipophilic balance (HLB) value, and critical micellar concentration (CMC). Instead, surfactants act in an all-encompassing mechanism, including surface electrochemistry, their solution and interfacial phases, and the chemical luminescence pathway. This leads to dramatic differences in signals obtained, ranging from 5-fold increases to total quenching. Within this complexity, we defined six guiding principles that are extrapolated from the underlying mechanisms and selection guides for surfactant, electrode, and environmental condition combinations. Those will now assist in developing highly sensitive luminol-ECL-based bioassays, because the surfactant selection can be based not only on properties needed for the assay protocol but also on identifying the optimal electrode-surfactant pair to maximize detection efficiency.


Subject(s)
Luminol/chemistry , Surface-Active Agents/chemistry , Adsorption , Electrochemical Techniques/instrumentation , Electrochemical Techniques/methods , Electrodes , Luminescence , Luminescent Measurements/methods
7.
Angew Chem Int Ed Engl ; 57(2): 408-411, 2018 01 08.
Article in English | MEDLINE | ID: mdl-29119667

ABSTRACT

The most efficient and commonly used electrochemiluminescence (ECL) emitters are luminol, [Ru(bpy)3 ]2+ , and derivatives thereof. Luminol stands out due to its low excitation potential, but applications are limited by its insolubility under physiological conditions. The water-soluble m-carboxy luminol was synthesized in 15 % yield and exhibited high solubility under physiological conditions and afforded a four-fold ECL signal increase (vs. luminol). Entrapment in DNA-tagged liposomes enabled a DNA assay with a detection limit of 3.2 pmol L-1 , which is 150 times lower than the corresponding fluorescence approach. This remarkable sensitivity gain and the low excitation potential establish m-carboxy luminol as a superior ECL probe with direct relevance to chemiluminescence and enzymatic bioanalytical approaches.


Subject(s)
Electrochemical Techniques/methods , Luminol/chemistry , Fluorescence , Limit of Detection , Liposomes , Luminescence , Solubility , Water
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