Subnanomolar Detection Limits Research Articles

Systematic evaluation of benzoylation for liquid chromatography-mass spectrometry analysis of different analyte classes

LC-MS is an indispensable tool for small molecule analysis in many fields; however, many small molecules require chemical derivatization to improve retention on commonly used reversed-phase columns and increase ionization. Benzoyl chloride (BzCl) derivatization is commonly used for derivatization of primary and secondary amines and phenolic alcohols, though evidence exists that with proper reaction conditions (i.e., specific bases), other hydroxyl groups may be derivatized too. Previous studies have examined BzCl concentration, reaction times, and reaction temperatures for derivatization of amines and phenols for LC-MS analysis; however, use of different bases, base concentration, and extending to conditions to hydroxyl groups for LC-MS analysis has not been well-studied. To address this understudied area and identify reaction conditions for both amino and hydroxyl groups, we performed a systematic study of reaction conditions on multiple classes of potential targets. For selected derivatization methods, detection limits and performance in a variety of biological matrices were assessed. Results highlight the importance of tailoring derivatization methods for a given application as they varied by molecule and/or molecule class. Compared to the standard BzCl method commonly used, alternative methods were identified to better derivatize challenging analytes (glucosamine, choline, cortisol, uridine, cytidine) with detection limits reaching 1100, 9, 38, 170, and 67 nM compared to undetectable, 170, 86, 1000, and 86 nM respectively. Sub-nanomolar detection limits were achieved for norepinephrine with alternative derivatization approaches. Improved derivatization methods for several classes and molecules including nucleosides, steroids, and molecules containing hydroxyl groups were also identified.

Portable smartphone-enabled dydrogesterone sensors based on biomimetic polymers for personalized gynecological care.

Dydrogesterone, a frequently prescribed synthetic hormone integral to the treatment of diverse gynecological conditions, necessitates precise quantification in complex human plasma. In this study, the development of a portable, smartphone-based electrochemical sensor employing screen-printed gold electrodes (SPAuEs) modified with a biomimetic, molecularly imprinted poly(methacrylic acid-co-methyl methacrylate) (MIP) is presented for dydrogesterone detection in human plasma. FTIR spectroscopy illustrates the transformation of a pre-polymer mixture into a polymerized matrix, while SEM reveals a uniform MIP/SPAuE surface morphology. The sensor fabrication protocol, encompassing MIP/SPAuE composition, polymerization solvent, incubation time, and scan rate, is optimized to achieve enhanced sensitivity. The MIP/SPAuEs sensor exhibits a linear sensor response to dydrogesterone within the concentration range of 1-500 nM, as evidenced by cyclic and differential pulse voltammetry. The MIP/SPAuE sensor demonstrates exceptional sensitivity, recording 8.2 × 10-3 μA nM-1, with a sub-nanomolar limit of detection (LOD = 370 pM), and low limit of quantification (LOQ = 1.12 nM), along with appreciable selectivity over common interferents. In real-world clinical applications, the designed sensor is effectively employed for the rapid and precise determination of dydrogesterone in human blood plasma, achieving a remarkable recovery of 81%. Furthermore, MIP/SPAuE coatings possess suitable stability over 15 days, indicating the robustness of the sensor material for multiple rounds of analysis. The developed sensor provides a sensitive, selective, and cost-effective solution for monitoring dydrogesterone in plasma during various gynecological disorders, allowing for personalized healthcare applications.

Discerning biomimetic nanozyme electrodes based on g-C3N4 nanosheets and molecularly imprinted polythiophene nanofibers for detecting creatinine in microliter droplets of human saliva

The growing risk of death associated with kidney dysfunction underlines the requirement for a cost-effective and precise point-of-care (POC) diagnostic tool to identify chronic kidney disease (CKD) at an early stage. This work reports the development of a non-invasive POC diagnostic based on cost-efficient, disposable electrodes and in situ-designed biomimetic nanozymes. The nanozymes are composed of graphitic carbon nitride nanosheets (gCN) and creatinine-imprinted polythiophene nanofibers (miPTh). Microscopic analyses reveal porous nanofibrous surface morphology of biomimetic miPTh/gCN nanozymes. Bulk imprinting and the inclusion of conductive gCN nanosheets drastically reduced the charge transfer resistance and improved the electron exchange kinetics at the nanozyme-electrolyte interface. The electrochemical oxidation of creatinine is studied via cyclic voltammetry (CV), and differential pulse voltammetry (DPV), which exhibit excellent creatinine recognition ability of biomimetic miPTh/gCN nanozyme sensors compared to pristine polymeric or non-imprinted nanozymes. The sensor reveals linear response toward 200–1000 nmol L−1 creatinine, high sensitivity (4.27 μA cm−2 nmol−1 L), sub-nanomolar detection limit (340 pmol L−1), and excellent selectivity over common salivary analytes. To corroborate its real-world utility, the miPTh/gCN nanozyme sensor shows an impressive 94.8% recovery of spiked creatinine concentrations in microliter droplets of human saliva samples. This disposable sensor reveals great potential in the realm of reliable and efficient non-invasive POC diagnostics for healthcare delivery.

Aptamer-enhanced particle aggregation inhibition assay for simple homogeneous protein detection using DNA aptamer and thermo-responsive magnetic nanoparticles

A simple and sensitive homogeneous protein detection system is required for the early detection of biomarkers. Thermo-responsive magnetic particles (TM) have already been developed to achieve easy bound/free separation at the homogeneous protein detection system, but they are still limited owing to the requirement of secondary antibodies and negatively charged polymers, and it is challenging to control the TM aggregation behavior because of the size of the TM. Therefore, at new method to control TM aggregation behavior that is simple, easy, and highly sensitive is required. In this study, we developed a DNA aptamer-based TM assay as a simple protein detection system without additional secondary molecular recognition elements or negatively charged polymer. In the first attempt, a DNA aptamer was modified on the TM surface, and its aggregation behavior was monitored depending on the target molecule concentration. The TM aggregation rate during the heating process decreased depending on the amount of the DNA aptamer and increased depending on the target protein level. This suggests that the DNA aptamer prevented TM aggregation owing to its negative charge and achieved target protein detection owing to the cancellation of repulsion. Capturable aptamers were used in the TM assay to improve the sensitivity and limit of detection. The designed Capture DNA was modified on the TM surface, and the aptamer was captured in the presence of the target protein through a conformational change. Eventually, Capturable aptamer-based TM assay achieved a sub-nanomolar limit of detection and higher sensitivity than that of our initial investigation. Through this study and the ease of the DNA aptamer design, it was shown that the DNA aptamer-modified TM assay enabled the development of a simple and sensitive homogeneous protein detection system.

A new eco-friendly electrochemical sensing platform for monitoring of agomelatine in different matrices with sub-nanomolar detection limits

The first eco-friendly, sensitive, and cost-effective electrochemical sensing platform for agomelatine monitoring in human plasma was developed. It was successfully applied also for agomelatine determination in urine and commercial tablets. Cobalt nanoparticles (Co-NPs) and sugar polymer (SP) were utilized in the fabrication of a modified carbon paste electrode (Co-NPs/SP/MCPE). Compared to a bare electrode, Co-NPs/SP/MCPE displayed significant enhancement in the surface area and the anodic peak current with decrease in charge transfer resistance across the electrode interface. Under the optimum conditions, Co-NPs/SP/MCPE showed good linearity over two concentration ranges 3.5 × 10−10 – 1.0 × 10−7 M and 2.0 × 10−7 – 1.0 × 10−4 M using square wave voltammetry. LOD and LOQ were found to be 2.12 × 10−10 and 6.44 × 10−10 M, respectively indicating excellent sensitivity. Good recoveries obtained in different matrices show that Co-NPs/SP/MCPE is a promising analytical tool for routine analysis and point-of-care diagnostics. The proposed sensor proved to be an excellent green analytical tool according to the analytical eco-scale for greenness assessment.

Quantum Dot-Peptide Conjugates as Energy Transfer Probes for Sensing the Proteolytic Activity of Matrix Metalloproteinase-14.

We detail the assembly and characterization of quantum dot (QD)-dye conjugates constructed using a peptide bridge specifically designed to recognize and interact with a breast cancer biomarker─matrix metalloproteinase-14 (MMP-14). The assembled QD conjugates are then used as optically addressable probes, relying on Förster resonance energy transfer (FRET) interactions as a transduction mechanism to detect the activity of MMP-14 in solution phase. The QDs were first coated with dithiolane poly(ethylene glycol) (PEG) bearing a carboxyl group that allows coupling via amide bond formation with different dye-labeled peptides. The analytical capability of the conjugates is enabled by correlating changes in the FRET efficiency with the conjugate valence and/or QD-to-dye separation distance, triggered and modulated by enzymatic proteolysis of surface-tethered peptides. The FRET probe exhibits great sensitivity to enzyme digestion with sub-nanomolar limit of detection. We further analyze the proteolysis data within the framework of the Michaelis-Menten model, which considers the fact that surface-attached peptides have a slower diffusion coefficient than free peptides. This results in reduced collision frequency and lower catalytic efficiency, kcat/KM. Our results suggest that our conjugate design is promising, effective, and potentially useful for in vivo analysis.

Highly sensitive and rapid detection of hypochlorous acid in aqueous media and its bioimaging in live cells and zebrafish using an ESIPT-driven mycophenolic acid-based fluorescent probe.

Excessive production of potent biological oxidants such as HOCl has been implicated in numerous diseases. Thus, it is crucial to develop highly specific and precise methods to detect HOCl in living systems, preferably with molecules that can show a distinct therapeutic effect. Our study introduces the synthesis and application of a highly sensitive fluorescence "turn-on" probe, Myco-OCl, based on the mycophenolic acid scaffold with exceptional water solubility. The ESIPT-driven mechanism enables Myco-OCl to specifically and rapidly detect (<5 s) HOCl with an impressive Stokes shift of 105 nm (λex = 417 nm, λem = 522 nm) and a sub-nanomolar (97.3 nM) detection limit with the detection range of 0 to 50 μM. The potential of Myco-OCl as an excellent biosensor is evident from its successful application for live cell imaging of exogenous and endogenous HOCl. In addition, Myco-OCl enabled us to detect HOCl in a zebrafish inflammatory animal model. These underscore the great potential of Myco-OCl for detecting HOCl in diverse physiological systems. Our findings thus offer a highly promising tool for detecting HOCl in living organisms.

Peptide-Induced Fractal Assembly of Silver Nanoparticles for Visual Detection of Disease Biomarkers.

We report the peptide-programmed fractal assembly of silver nanoparticles (AgNPs) in a diffusion-limited aggregation (DLA) mode, and this change in morphology generates a significant color change. We show that peptides with specific repetitions of defined amino acids (i.e., arginine, histidine, or phenylalanine) can induce assembly and coalescence of the AgNPs (20 nm) into a hyperbranched structure (AgFSs) (∼2 μm). The dynamic process of this assembly was systematically investigated, and the extinction of the nanostructures can be modulated from 400 to 600 nm by varying the peptide sequences and molar ratio. According to this rationale, two strategies of SARS-CoV-2 detection were investigated. The activity of the main protease (Mpro) involved in SARS-CoV-2 was validated with a peptide substrate that can bridge the AgNPs after the proteolytic cleavage. A sub-nanomolar limit of detection (0.5 nM) and the capacity to distinguish by the naked eye in a wide concentration range (1.25-30 nM) were achieved. Next, a multichannel sensor-array based on multiplex peptides that can visually distinguish SARS-CoV-2 proteases from influenza proteases in doped human samples was investigated.

Rational design of a mesoporous silica@ZIF-8 based molecularly imprinted electrochemical sensor with high sensitivity and selectivity for atropine monitoring

Porous materials are of great interest for sensor design, while poor conductivity and single structure hinder their electrochemical application seriously, and the development of an effective way to improve their specificity for targets is another challenge. Herein, novel flower-like core-shell mesoporous silica [email protected] imidazolate framework-8 ([email protected]) hybrid electrocatalyst with hierarchical framework, enhanced porosity, tunable functionality, and excellent stability was firstly attempted. Significantly, the electronic environment and dynamics were greatly enhanced in [email protected], owing to the abundant faradic active sites of MSN inner cores and plentiful pores for electrolyte permeation and analytes adsorption of ZIF-8 outer shell. Thereafter, the molecularly imprinted electrochemical sensor based on [email protected] nanocomposites was prepared with p-aminothiophenol as functional monomer, Au nanoparticles as cross-linker, and atropine as template molecule via electropolymerization technique, which had a wide linear range, subnanomolar detection limit, and specific selectivity for atropine monitoring. Our work will inspire further studies on developing porous electrocatalysts with improved stability and performance, which is highly expected for varieties of sensor applications.

Expanding the codes: The development of density-encoded hydrogel microcarriers for suspension arrays

Although suspension array technology (SAT), which uses encoded microspheres, provides high-quality results with versatile applicability for information-intensive bioanalytic applications, current encoding strategies limit the number of codes that can be distinguished. In this paper, we introduce density-encoded hydrogel microcarriers (DMs), which employ the intrinsic density property of biomaterials as a high-capacity coding dimension. Two hydrogel monomers were employed at different ratios to synthesize microgels with distinctive densities. DMs not only can be simultaneously decoded and separated using density gradient centrifugation, but also are compatible with flow cytometry detection. The size and color of DMs have been used as extra coding parameters, to construct an 8 × 2 × 4 (density × size × color) three-dimensionally encoded hydrogel microcarrier library. With aptamer-functionalized DMs (ADMs), we developed a 4-plex protein quantification method for the label-free detection of plasma biomarkers with sub-nanomolar detection limits and good linearities. Moreover, ADMs can be used for label-free naked-eye detection of tumor-derived exosomes. We believe that the simplicity and functionality of DMs will advance the field of suspension arrays and inspire the development of DM-based diagnostic applications.

In situ monitoring of triclosan in environmental water with subnanomolar detection limits using eco-friendly electrochemical sensors modified with cyclodextrins.

The environmental emergence of unexpected contaminants has gained the attention of the scientific community. A broad spectrum antimicrobial compound named triclosan (TCS) was detected in the environment as an emerging contaminant. Owing to its inherent toxicity, we have proposed eco-friendly potentiometric liquid state sensors to be used for monitoring and quantifying TCS in environmental water samples. The proposed sensors have been optimized by modifying the inner filling solution using hydrophilic 2-hydroxypropyl β-cyclodextrin as a complexing agent to be capable of minimizing the trans-membrane ion flux and hence improving the selective and sensitive determination of TCS in environmental matrices with low LOD values. The obtained linear response of the optimized sensor was (1 × 10-9 to 1 × 10-5 M) compared to the control sensor (1 × 10-8 to 1 × 10-4 M). The obtained limit of detection (LOD) value was found to be 9.86 × 10-10 M compared to 9.78 × 10-9 M of the control sensor. The modification of the inner filling solution of the sensor with 2-hydroxypropyl β-cyclodextrin improves not only its sensitivity but also its response time to be only 5 seconds. The electrical performance of the proposed sensor was evaluated following IUPAC recommendations. Both the pH and temperature effects were studied and optimized. Two different greenness assessment tools, Analytical Eco-scale and Green Procedure Index, were adopted upon the evaluation of the proposed sensors' greenness.

Grafting of reactive dyes onto lignocellulosic material: Application for Pb(II) adsorption and electrochemical detection in aqueous solution

Triplochyton scleroxylon (Ayous) sawdust was modified by grafting two reactive dyes (Reactive Red 120 (RR120) and Reactive Black 5 (RB5)) inspired by the strategy used for cellulosic fibers dyeing. The visual appearance as well as the infrared spectra and the point of zero charge determination confirm the effective attachment of the dyes onto sawdust. The anionic charges resulting from the grafted dyes were exploited for Pb(II) biosorption and electrochemical detection in aqueous solution. The presence of dyes grafted onto Ayous sawdust (SA) greatly improves its affinity for Pb(II) biosorption as confirmed by the increase of the pseudo-second order kinetic rate constant, especially in the case of RR120 modified sawdust (SA-RR120). Moreover, the maximum adsorption capacities obtained from the Langmuir model (207 µmol g−1, 281 µmol g−1 and 298 µmol g−1 respectively for SA, SA-RB5 and SA-RR120) confirmed the improvement in adsorption resulting from sawdust dyeing. Used as glassy carbon electrode (GC) modifiers, the functionalized sawdust improved the electrochemical detection of Pb(II). After optimization of the experimental parameters, sub-nanomolar detection limits were obtained (1.3 nM, 0.7 nM and 0.4 nM for GC/SA, GC/SA-RB5 and GC/SA-RR120 respectively). In addition to such improvements in Pb(II) biosorption and trace-level detection, these materials also showed appreciable selectivity and excellent performance in real media samples.

Newly designed gel-integrated nanostructured gold-based interconnected microelectrode arrays for continuous in situ arsenite monitoring in aquatic systems

This work presents the development, characterization and field validation of newly designed gel-integrated nanostructured gold-basedmicroelectrode arrays (Au-GIMEs) enabling the direct detection of inorganic arsenite (As(III)) in fresh and marine aquatic systems. They consist of renewable Au nanoparticles (AuNP) or Au nanofilaments (AuNF) electroplated on 100- to 500- interconnected iridium (Ir)-based microdisk arrays and covered with a gel. The gel protects the sensor surface from fouling and ensures that mass transport of analytes toward the sensor surface is by diffusion only, and therefore independent of the ill-controlled hydrodynamic conditions of the media. The responses of these sensors to direct Square Wave Anodic Stripping Voltammetry (SWASV) quantification of As(III) at pH 7.6 were investigated first in 0.1 M NaNO3, then in fresh and marine water samples. The analytical responses were found to be correlated to the number of interconnected microelectrodes and the morphology of the nanostructured Au deposits but independent of the media composition. The new interconnected AuNF-GIME have sub-nanomolar detection and quantification limits fulfilling the requirement for direct monitoring of As(III) in fresh and marine aquatic systems. The AuNF-GIME were incorporated in a submersible multi-channel trace metal sensing probe for remote high-resolution monitoring. Field evaluation and validation was performed during a one-week field study in the Elbe Estuary (North Germany), from which environmental data are presented.

Ultratrace Metal Speciation Analysis by Coupling of Sector-Field ICP-MS to High-Resolution Size Exclusion and Reversed-Phase Liquid Chromatography.

Techniques for metal speciation analysis with subnanomolar (ppt) detection limits in complex matrices, with simultaneous quantification of matrix elements, have become a necessity for investigating targets of trace metal binding to macromolecules and pigments at environmentally relevant concentrations. In this work we optimized the analysis of such metal binding in a custom-built HPLC-ICP-MS system. Key elements of the optimization were the choice of components for the metal-free HPLC-DAD system and sector-field ICP-MS detection (ICF-sfMS) with desolvating injection and optimization of sample handling. Protein analysis was done using ammonium bicarbonate buffer and size exclusion chromatography (SEC-ICP-sfMS), with possible addition of anion exchange chromatography. Detection of metal exchange in pigments (chlorophylls and bacteriochlorophylls) was based on reversed-phase chromatography with a methanol-acetone gradient and coupling to the ICP-sfMS via a dedicated organic matrix interface (RPC-ICP-sfMS). The resulting HPLC-DAD-ICP-sfMS system has detection limits in the picomolar range in protein buffer, limited by the maximal achievable purity of buffers/solvents and not by system sensitivity. Tests for method optimization showed that sonication, meant to increase protein solubilization, leads to artifacts of metal loss from metalloproteins. Examples for Cd binding to soybean proteins and chlorophyll, Cr binding to Arabidopsis thaliana proteins, La binding to Desmodesmus quadricauda proteins, and Cu binding to Rhodospirillum rubrum proteins and pigments are shown. These application examples demonstrate that the system is sensitive enough to detect binding of metals to proteins and pigments at background concentration levels of typical nutrient solutions made from analytical grade chemicals, equivalent to ultratrace metal concentrations in nonpolluted environments.

Liposome-Enhanced Polymerization-Based Signal Amplification for Highly Sensitive Naked-Eye Biodetection in Paper-Based Sensors.

Polymerization-based signal amplification (PBA) is a material-based approach to improving the sensitivity of paper-based diagnostic tests. Eosin Y is used as an assay label to photo-initiate free-radical polymerization to produce colored hydrogels in the presence of target analytes captured by bioactive paper. PBA achieves high-contrast and time-independent signals, but its nanomolar detection limit makes it impractical for early diagnosis of many diseases. In this work, we demonstrated efficient localization of large quantities of eosin Y per captured target analyte by incorporating eosin Y-loaded liposomes into PBA. This new "materials approach" allowed 30-fold signal enhancement compared to conventional PBA. To further improve the detection limit of liposome-enhanced PBA, we used a continuous flow-through assay format with 100 μL of analyte solution, achieving sub-nanomolar detection limits with high-contrast signals that were easily discernible to the unaided eye.

Electrochemical Immuno- and Aptasensors for Mycotoxin Determination

Modern analysis of food and feed is mostly focused on development of fast and reliable portable devices intended for field applications. In this review, electrochemical biosensors based on immunological reactions and aptamers are considered in the determination of mycotoxins as one of most common contaminants able to negatively affect human health. The characteristics of biosensors are considered from the point of view of general principles of bioreceptor implementation and signal transduction providing sub-nanomolar detection limits of mycotoxins. Moreover, the modern trends of bioreceptor selection and modification are discussed as well as future trends of biosensor development for mycotoxin determination are considered.

DNA-Templated In Situ Synthesis of Highly Dispersed AuNPs on Nitrogen-Doped Graphene for Real-Time Electrochemical Monitoring of Nitric Oxide Released from Live Cancer Cells.

Dispersion promotion of nanomaterials can significantly enhance their catalytic activities. With a new DNA-templated in situ synthesis approach, we report the preparation of highly dispersed AuNPs on nitrogen-doped graphene sheets (NGS) with significantly improved electrocatalytic ability for the monitoring of nitric oxide (NO) released from live cancer cells. The template DNA is adsorbed on NGS via π-π stacking, and the Au precursor chelates along the DNA lattice through dative bonding. Subsequent introduction of the reducing agent leads to in situ nucleation and growth of AuNPs, eventually resulting in highly dispersed AuNPs on NGS. Because of the synergistic enhancement of the catalytic activities of AuNPs and NGS, as well as the high dispersion of AuNPs, such a nanocomposite shows significant electro-oxidation capability toward NO, leading to a highly sensitive subnanomolar detection limit for NO in vitro. More importantly, the laminin glycoproteins can be readily adsorbed on the surface of the nanomaterials to render excellent biocompatibility for the adhesion and proliferation of live cells, enabling the biointerface for electrochemical detection of NO released from live cancer cells.

Affinity Biosensors for Detection of Mycotoxins in Food.

This chapter reviews recent achievements in methods of detection of mycotoxins in food. Special focus is on the biosensor technology that utilizes antibodies and nucleic acid aptamers as receptors. Development of biosensors is based on the immobilization of antibodies or aptamers onto various conventional supports like gold layer, but also on nanomaterials such as graphene oxide, carbon nanotubes, and quantum dots that provide an effective platform for achieving high sensitivity of detection using various physical methods, including electrochemical, mass sensitive, and optical. The biosensors developed so far demonstrate high sensitivity typically in subnanomolar limit of detection. Several biosensors have been validated in real samples. The sensitivity of biosensors is similar and, in some cases, even better than traditional analytical methods such as ELISA or chromatography. We believe that future trends will be focused on improving biosensor properties toward practical application in food industry.

On-site preconcentration of pesticide residues in a drop of seawater by using electrokinetic trapping, and their determination by surface-enhanced Raman scattering.

The authors have developed a method for simultaneous quantification of several charged pesticides (as shown for amitrole, simazine, trichlorfon and bisultap). It is based on the use of a reduced graphene oxide-modified screen-printed electrode (RGO-SPE) and combines electrokinetic trapping (EKT) and surface-enhanced Raman scattering (SERS). When a 50μL droplet containing negatively charged RGO and positively charged gold nanorods is placed on the SPE, the RGO and gold nanorods are selectively attracted on the surface of the SPE during EKT. This leads to the formation of sandwich-type hybrid substrates. The resulting substrates also contain Raman "hot spots" among the high-density gold nanorods. This, along with the excellent adsorption performance of RGO, makes it an excellent SERS substrate for on-site detection of the charged pesticides. The method is highly reproducible and long-term stable. The spot-to-spot variation of the intensity of the SERS is <15%, and the performance of SERS activity is maintained over a period of 6weeks. The method works over a wide range of concentrations (0.5nM to 4μM) for charged pesticides under optimal conditions, with a sub-nanomolar detection limit (at a signal to noise ratio of 3). The EKT-SERS method requires only microliter volumes and takes only minutes for completion. Therefore, the method provides high sensitivity for detection while preserving the selectivity and stability required for reliable quantitative analysis. Graphical abstract A method combining electrokinetic trapping and SERS can be used for simultaneous detection of charged pesticides in a drop of seawater.

A Rayleigh surface acoustic wave (R-SAW) resonator biosensor based on positive and negative reflectors with sub-nanomolar limit of detection

A label-free sub-nanomolar Rayleigh surface acoustic wave (R-SAW)-based resonator biosensor is demonstrated for biomolecular detection in liquid after drying. The biosensor comprises two interdigital transducers for R-SAW generation and two positive and negative reflectors to confine the acoustic energy in the sensitive area. We benchmark this biosensor against biotin-streptavidin binding, which is a standard, well-known model for a variety of biosensing processes. The experiments demonstrate a limit of detection of 104pM and a normalized sensitivity of −296m2kg−1. As a comparison with similar acoustic-wave based systems, both sensitivity and limit of detection are better than that of standard commercial gravimetric sensors (i.e., quartz-crystal-microbalances) and generally better than that of more common Love-SAW biosensors. Our biosensor has a dynamic range potentially compatible with several health- and safety-related assays, among all cancer biomarker detection.