Detection Of C-reactive Protein Research Articles

Polyaromatic Hydrocarbon-Functionalized 2D MXene-Based 3D Porous Antifouling Nanocomposite with Long Shelf Life for High-Performance Electrochemical Immunosensor Applications.

Affinity-based electrochemical (AEC) biosensors have gained more attention in the field of point-of-care management. However, AEC sensing is hampered by biofouling of the electrode surface and degradation of the antifouling material. Therefore, a breakthrough in antifouling nanomaterials is crucial for the fabrication of reliable AEC biosensors. Herein, for the first time, we propose 1-pyrenebutyric acid-functionalized MXene to develop an antifouling nanocomposite to resist biofouling in the immunosensors. The nanocomposite consisted of a 3D porous network of bovine serum albumin cross-linked with glutaraldehyde with functionalized MXene as conductive nanofillers, where the inherited oxidation resistance property of functionalized MXene improved the electrochemical lifetime of the nanocomposite. On the other hand, the size-extruded porous structure of the nanocomposite inhibited the biofouling activity on the electrode surface for up to 90 days in real samples. As a proof of concept, the antifouling nanocomposite was utilized to fabricate a multiplexed immunosensor for the detection of C-reactive protein (CRP) and ferritin biomarkers. The fabricated sensor showed good selectivity over time and an excellent limit of detection for CRP and ferritin of 6.2 and 4.2 pg/mL, respectively. This research successfully demonstrated that functionalized MXene-based antifouling nanocomposites have great potential to develop high-performance and low-cost immunosensors.

Electrochemical biosensor based on RNA aptamer and ferrocenecarboxylic acid signal probe for C-reactive protein detection

Monitoring health-related biomarkers using fast and facile detection techniques provides key physicochemical information for disease diagnosis or reflects body health status. Among them, electrochemical detection of various bio-macromolecules, e.g., the C-reactive protein (CRP), is of great interest in offering potential diagnosis for acute inflammation caused by infections, heart diseases, etc. Herein, a novel electrochemical aptamer biosensor was constructed from Ti3C2Tx MXene and in-situ reduced Au NPs for thiolated-RNA aptamer immobilization and CRP protein detection using Fc(COOH) as the signal probe. The sensory performances for CRP detection were optimized based on working conditions, including the incubation times and the pH. The large surface area offered by Ti3C2Tx MXene and high electrical conductivity originating from Au NPs endowed the as-fabricated aptamer biosensor with a decent sensitivity for CRP in a wide linear range of 0.05–80.0 ng/mL, good selectivity over interfering substances, and a low detection limit of 0.026 ng/mL. Such aptamer biosensors also detected CRP in serum samples using the spike & recovery method with reasonable recovery rates. The results demonstrated the potential of the as-fabricated electrochemical aptamer biosensor for fast and facile CRP detection in practical applications.

Label-free electrochemical detection of glycated hemoglobin (HbA1c) and C-reactive protein (CRP) to predict the maturation of coronary heart disease due to diabetes

The pathophysiological link between diabetes and heightened propensity for the development of coronary heart disease (CHD) is well-established. Prevailing evidence confirms that small increases in low concentrations of high-sensitivity C reactive protein (hs-CRP) in the human body can determine the tendency of developing CHD. Additionally, glycated hemoglobin (HbA1c) is a well-recognized biomarker to evaluate diabetes progression. Given the positive correlation between diabetes and CHD, this research presents a notably unprecedented label-free electrochemical approach for the dual detection of %HbA1c regarding Total Hb and hs-CRP, facilitating early CHD prediction and cost-effective point-of-care diagnostics. Furthermore, a novel redox probe O-(4-Nitrophenylphosphoryl)choline (C11H17N2O6P) was used for the electrochemical detection of CRP, a method not documented in scientific literature before. The calibration curves demonstrate a limit of detection (LOD) of 5 mg/mL in PBS (pH 8) and 6 mg/mL in simulated blood (SB) for a linear range of 0–30 mg/mL of HbA1c. Conjointly, a LOD of 0.007 mg/mL and 0.008 mg/mL for measurement in PBS (pH 7.4) and SB are reported for a linear range of 0–0.05 mg/mL of CRP. The electrochemical systems presented could accurately quantify HbA1c and CRP in mixed samples, demonstrating reasonable specificity and practical applicability for complex biological samples.

Aptamer-conjugated gold nanoparticles for portable, ultrasensitive naked-eye detection of C-reactive protein based on the Tyndall effect

BackgroundC-reactive protein (CRP) represents an early clinical biomarker that indicates the presence of inflammatory or infectious conditions in the human body. Today's procedures approved by the Food and Drug Administration (FDA) imply expensive equipment and highly trained personnel to perform the test. Therefore, a new diagnostic method with high detection efficiency and less cost is urgently needed for delivering rapid and timely results in point-of-care (POC) service. ResultsHerein, we propose a new, equipment-free, and portable sensing method for the future POC detection of CRP based on the Tyndall effect (TE). In our study, aptamer-conjugated citrate-stabilized gold nanoparticles (apta-AuNPs) are exploited as the sensing platform. The apta-AuNPs’ interaction with CRP in a saline environment leads to their aggregation, thus enhancing the scattering of light when the solution is exposed to a 640 nm pointer laser line. Firstly, the enhancement of the scattering light as a function of increasing concentration of CRP in solution is measured spectroscopically using a typical 90-degree angle spectrofluorometer and then the measurements are compared to the classic colorimetric detection using an UV–Vis spectrophotometer. Finally, to achieve high portability and accessibility, we demonstrate that the measurement of CRP concentration can be performed with similar accuracy but in a more direct and inexpensive way by using a laser pointer pen as the excitation source and a camera of a low-budget smartphone as a quantitative reader instead of most expensive spectrofluorometer. SignificanceThe portable TE-based assay exhibits a wide linear dynamic range (1–60 μg/mL) for the detection of CRP with a limit of detection (LOD) of 92 ng/mL The proposed method is capable to integrate both standard and high-sensitivity CRP analysis in a single procedure with increased sensitivity and prompt delivery of analysis results. Moreover, the sensing procedure is significantly faster than the FDA approved ones with a detection time of only 10 min. Finally, as a proof-of-concept, our findings demonstrate excellent recovery for CRP detection in spiked and diluted urine samples, highlighting the strong potential of this sensing method for POC applications.

Development and performance of NLISA for C-reactive protein detection based on Prussian blue nanoparticle conjugates.

Prussian blue nanoparticles (PBNPs), also called nanozymes, are very attractive as an alternative to horseradish peroxidase in immunoassay development due to their simple and low-cost synthesis, stability and high catalytic activity. Today, there is a method for highly effective PBNP synthesis based on the reduction of an FeCl3/K3[Fe(CN)6] mixture by hydrogen peroxide. However, there is a lack of research showcasing the use of these highly effective PBNPs for specific target detection in clinical settings, as well as a lack of comprehensive comparisons with conventional methods. To address this gap, we prepared diagnostic reagents based on highly effective PBNPs by modifying them using gelatin and attaching anti-C-reactive protein (CRP) monoclonal antibodies through cross-linking with glutaraldehyde. As a result, a solid-phase colorimetric immunoassay in a sandwich format (nanozyme-linked immunosorbent assay [NLISA]) using highly effective PBNPs as a label for CRP detection has been demonstrated for the first time. The assay demonstrated a detection limit of 21.8pg/mL, along with acceptable selectivity, precision (CV < 25%) and accuracy (the recovery index was within acceptable limits (75-125%) for LLOQ /ULOQ range. The analytical performance of this method is on par with sensitive assays developed in the last 5 years. Notably, the results obtained from NLISA align with those from an immunofluorescence assay conducted by a certified clinical laboratory. Furthermore, this study underscores the technological challenges involved in constructing an analysis that necessitate further exploration.

Scorpion-Inspired, Hydrophobic, Highly Sensitive, and Paper-Based Magnetoelastic Biosensor for C-Reactive Protein Detection

Scorpion-Inspired, Hydrophobic, Highly Sensitive, and Paper-Based Magnetoelastic Biosensor for C-Reactive Protein Detection

Magnetic Polystyrene Nanoparticles Prepared by Emulsion Solvent-Evaporation for the Chemiluminescent Immunoassay.

In clinical diagnosis, magnetic polystyrene nanoparticles (MPS NPs) are commonly applied to, e.g., the chemiluminescent immunoassay (CLEIA). However, the conventional preparation method of MPS NPs requires a long duration of heating to form polymer particles, which is inefficient. In this study, we prepared MPS NPs by emulsion solvent-evaporation without heating. We evaluated the effect of the solvent in the water and organic phases on the magnetic particle content. MPS NPs prepared by 4% (v/v) MeOH aqueous solution and adding stearic acid (SA) (4MeSA-MPS NPs) exhibited the highest magnetic particle content. Furthermore, CLEIA analysis indicates that the C-reactive protein detection limit is 80 pg/mL. Thus, 4MeSA-MPS NPs are promising for clinical diagnoses.

Graphene Field-Effect Transistors with Surface-Charge Modulation for C-Reactive Protein Detection in Artificial Saliva

Graphene Field-Effect Transistors with Surface-Charge Modulation for C-Reactive Protein Detection in Artificial Saliva

Development of a double antibody sandwich ELISA method for the quantitative detection of serum C-reactive protein based on nanobody

In this study, we successfully developed a nanobody-based double antibody sandwich ELISA kit for the detection of clinical serum C-reactive protein (CRP) by using two novel CRP specific nanobodies.The developed method exhibited a linear detection range of approximately 6–200 ng/mL, with a detection limit of 1 ng/mL. Furthermore, the method demonstrated excellent specificity, as there was no cross-reactivity with interfering substances such as total bilirubin and hemoglobin and so on. To assess reproducibility, independent measurements of the samples were conducted under experimental conditions, resulting in intra- and inter-batch coefficients of variation below 10% and a recovery rate of 93%–102%. These results indicate robust reproducibility of the method.To evaluate the performance of the developed kit, we collected 90 clinical samples for correlation analysis with commercial kits. The results showed a high correlation coefficient value (R2) of 0.98, indicating accurate concordance between the developed and commercial kits.In conclusion, our study successfully developed a nanobody-based double antibody sandwich ELISA kit to detect clinical serum CRP. The utilization of nanobodies represents a significant advancement in the field of CRP immunoassay development. The developed kit demonstrates excellent performance characteristics and holds promise for clinical applications.

Structure-Switching Electrochemical Aptasensor for Rapid, Reagentless, and Single-Step Nanomolar Detection of C-Reactive Protein.

C-reactive protein (CRP) is an acute-phase reactant and sensitive indicator for sepsis and other life-threatening pathologies, including systemic inflammatory response syndrome. Currently, clinical turn-around times for established CRP detection methods take between 30 min to hours or even days from centralized laboratories. Here, we report the development of an electrochemical biosensor using redox probe-tagged DNA aptamers, functionalized onto inexpensive, commercially available screen-printed electrodes. Binding-induced conformational switching of the CRP-targeting aptamer induces a specific and selective signal-ON event, which enables single-step and reagentless detection of CRP in as little as 1 min. The aptasensor limit of detection spans approximately 20-60 nM in 50% human serum with dynamic response windows spanning 1-200 or 1-500 nM (R = 0.97/R = 0.98 respectively). The sensor is stable for at least 1 week and can be reused numerous times, as judged from repeated real-time dosing and dose-response assays. By decoupling binding events from the signal induction mechanism, structure-switching electrochemical aptamer-based sensors provide considerable advantages over their adsorption-based counterparts. Our work expands on the retinue of such sensors reported in the literature and is the first instance of structure-switching electrochemical aptamer-based sensors (SS-EABs) for reagentless, voltammetric CRP detection. We hope this study inspires further investigations into the suitability of SS-EABs for diagnostics, which will aid translational R&D toward fully realized devices aimed at point-of-care applications or for broader use by the public.

Rapid and Convenient Single-Chain Variable Fragment-Employed Electrochemical C-Reactive Protein Detection System.

Although IgG-free immunosensors are in high demand owing to ethical concerns, the development of convenient immunosensors that alternatively integrate recombinantly produced antibody fragments, such as single-chain variable fragments (scFvs), remains challenging. The low affinity of antibody fragments, unlike IgG, caused by monovalent binding to targets often leads to decreased sensitivity. We improved the affinity owing to the bivalent effect by fabricating a bivalent antibody-enzyme complex (AEC) composed of two scFvs and a single glucose dehydrogenase, and developed a rapid and convenient scFv-employed electrochemical detection system for the C-reactive protein (CRP), which is a homopentameric protein biomarker of systemic inflammation. The development of a point-of-care testing (POCT) system is highly desirable; however, no scFv-based CRP-POCT immunosensors have been developed. As expected, the bivalent AEC showed higher affinity than the single scFv and contributed to the high sensitivity of CRP detection. The electrochemical CRP detection using scFv-immobilized magnetic beads and the bivalent AEC as capture and detection antibodies, respectively, was achieved in 20 min without washing steps in human serum and the linear range was 1-10 nM with the limit of detection of 2.9 nM, which has potential to meet the criteria required for POCT application in rapidity, convenience, and hand-held detection devices without employing IgGs.

Multiplexed Capillary-Flow Driven Immunoassay for Respiratory Illnesses.

Multiplexed analysis in medical diagnostics is widely accepted as a more thorough and complete method compared to single-analyte detection. While analytical methods like polymerase chain reaction and enzyme-linked immunosorbent assay (ELISA) exist for multiplexed detection of biomarkers, they remain time-consuming and expensive. Lateral flow assays (LFAs) are an attractive option for point-of-care testing, and examples of multiplexed LFAs exist. However, these devices are limited by spatial resolution of test lines, large sample volume requirements, cross-reactivity, and poor sensitivity. Recent work has developed capillary-flow microfluidic ELISA platforms as a more sensitive alternative to LFAs; however, multiplexed detection on these types of devices has yet to be demonstrated. In the aftermath of the initial SARS-CoV-2 pandemic, the need for rapid, sensitive point-of-care devices has become ever clearer. Moving forward, devices that can distinguish between diseases with similar presenting symptoms would be the ideal home diagnostic. Here, the first example of a multiplexed capillary-flow immunoassay device for the simultaneous detection of multiple biomarkers is reported. From a single sample addition step, the reagents and washing steps required for two simultaneous ELISAs are delivered to spatially separated test strips. Visual results can be obtained in <15 min, and images captured with a smartphone can be analyzed for quantitative data. This device was used to distinguish between and quantify H1N1 hemagglutinin (HA) and SARS-CoV-2 nucleocapsid protein (N-protein). Using this device, analytical detection limits of 840 and 133 pg/mL were obtained for hemagglutinin and nucleocapsid protein, respectively. The presence of one target in the device did not increase the signal on the other test line, indicating no cross-reactivity between the assays. Additionally, simultaneous detection of both N-protein and HA was performed as well as simultaneous detection of N-protein and human C-reactive protein (CRP). Elevated levels of CRP in a patient infected with SARS-CoV-2 have been shown to correlate with more severe outcomes and a greater risk of death as well. To further expand on the simultaneous detection of two biomarkers, CRP and N-protein were detected simultaneously, and the presence of SARS-CoV-2 N-protein did not interfere with the detection of CRP when both targets were present in the sample.

Fabrication and sensing properties of a molecularly imprinted polymer on a photonic PDMS substrate for the optical detection of C-reactive protein

This work presents the development of a novel photonic-based sensor on a nanoscale patterned polydimethylsiloxane (PDMS) substrate acting as transducer, combined with a molecularly imprinted polymer for the rapid and sensitive detection of C-reactive protein (CRP). A silicon wafer was patterned using electron-beam lithography with a one-dimensional (1D) reflective grating (periodicity of about 400 nm) and then replicated using nanoimprint lithography. The latter silicon mould was used for replica moulding to generate a patterned PDMS. The unique 1D design gave PDMS photonic properties that were suitable as a substrate to assemble the biorecognition layer. The surface of the photonic PDMS was modified to become hydrophilic and additionally functionalized with vinyl silane groups to bind the imprinted polymer, resulting in a photonic molecularly imprinted polymer substrate (PMIPS). The developed method was successfully used to detect CRP in serum samples. The reflectance intensity decreased with increasing CRP concentrations, and the sensing PMIPS showed a linear response between 0.005 and 1.215 µg mL−1, a limit of detection of 0.003 µg mL−1, and a selective response to CRP when tested against serum calprotectin. Overall, this novel optical sensor can be used to detect CRP, a serum biomarker of inflammation, particularly in inflammatory bowel diseases.

Highly Sensitive, Stable InP Quantum Dot Fluorescent Probes for Quantitative Immunoassay Through Nanostructure Tailoring and Biotin-Streptavidin Coupling.

Nontoxic, highly sensitive InP quantum dot (QD) fluorescent immunoassay probes are promising biomedical detection modalities due to their unique properties. However, InP-based QDs are prone to surface oxidation, and the stability of InP QD-based probes in biocompatible environments remains a crucial challenge. Although the thick shell can provide some protection during the phase transfer process of hydrophobic QDs, the photoluminescence quantum yield (PLQY) is generally decreased because of the contradiction between lattice stress relaxation and thick shell growth. Herein, we developed thick-shell InP-based core/shell QDs by inserting a ZnSeS alloy layer. The ternary ZnSeS intermediate shell could effectively facilitate lattice stress relaxation and passivate the defect states. The synthesized InP/ZnSe/ZnSeS/ZnS core/alloy shell/shell QDs (CAS-InP QDs) with nanostructure tailoring revealed a larger size, high PLQY (90%), and high optical stability. After amphiphilic polymer encapsulation, the aqueous CAS-InP QDs presented almost constant fluorescence attenuation and stable PL intensity under different temperatures, UV radiation, and pH solutions. The CAS-InP QDs were excellent labels of the fluorescence-linked immunosorbent assay (FLISA) for detecting C-reactive protein (CRP). The biotin-streptavidin (Bio-SA) system was first introduced in the FLISA to further improve the sensitivity, and the CAS-InP QDs-based SA-Bio sandwich FLISA realized the detection of CRP with an impressive limit of detection (LOD) of 0.83 ng/mL. It is believed that the stable and sensitive InP QD fluorescent probes will drive the rapid development of future eco-friendly, cost-effective, and sensitive in vitro diagnostic kits.

Multiplex Assay of Cytokines with Tunable Detection Ranges for the Precise Diagnosis of Breast Cancer.

Precise profiling of the cytokine panel consisting of different levels of cytokines can provide personalized information about several diseases at certain stages. In this study, we have designed and fabricated an "all-in-one" diagnostic tool kit to bioassay multiple inflammatory cytokines ranging from picograms per milliliter to μg/mL in a small cytokine panel. Taking advantage of the kit fabricated by the DNA-encoded assembly of nanocatalysts in dynamic regulation and signal amplification, we have demonstrated the multiplex, visual, and quantitative detection of C-reactive protein (CRP), procalcitonin (PCT), and interleukin-6 (IL-6) with limits of detection of 1.6 ng/mL (61.54 pM), 20 pg/mL (1.57 pM), and 4 pg/mL (0.19 pM), respectively. This diagnostic tool kit can work well with commercial kits for detecting serum cytokines from breast cancer patients treated with immunotherapies. Furthermore, a small cytokine panel composed of CRP, PCT, and IL-6 is revealed to be significantly heterogeneous in each patient and highly dynamic for different treatment courses, showing promise as a panel of quantitative biomarker candidates for individual treatments. So, our work may provide a versatile diagnostic tool kit for the visual detection of clinical biomarkers with an adjustable broad detection range.

Multiplex signal amplification for ultrasensitive CRP assay via integrated electrochemical biosensor array using MOF-derived carbon material and aptamers

Accurate and precise detection of disease-associated proteins, such as C-reactive protein (CRP), remains a challenge in biosensor development. Herein, we present a novel approach－an integrated disposable aptasensor array－designed for precise, ultra-sensitive, and parallel detection of CRP in plasma samples. This integrated biosensing array platform enables multiplex parallel testing, ensuring the accuracy and reliability in sample analysis. The ultra-sensitivity of this biosensor is achieved through multiplex signal amplification. Leveraging the superior conductivity and extensive surface area of MOF-derived nanoporous carbon material (CMOF), the biosensor enhances recognition elements (aptamers) by catalyzing the horseradish peroxidase (HRP) label enzyme reaction to multiply the number of probe molecules. Optimized conditions yielded exceptional performance, exhibiting high accuracy (relative standard deviation, RSD≤10.0 %), a low detection limit (0.3 pg/mL, S/N = 3), ultra-sensitivity (0.16 μA/ng mL−1 mm−2), and a rapid response (seven parallel tests within 60 min). Importantly, this multi-unit integrated disposable aptasensor array accurately quantified CRP in human serum, demonstrating comparable results to commercial enzyme–linked immunosorbent assay (ELISA). This technology showcases promise for detecting various biomarkers using a unified approach, presenting an appealing strategy for early disease diagnosis and biological analysis.

Three-Dimensional Printed Flexible Magnetoelastic Biosensor for C-Reactive Protein Detection Based on Patterned PDMS/MnFe2O4 Film

Three-Dimensional Printed Flexible Magnetoelastic Biosensor for C-Reactive Protein Detection Based on Patterned PDMS/MnFe₂O₄ Film

Mesoporous silica-supported platinum nanocatalysts for colorimetric detection of glucose, cholesterol, and C-reactive protein.

Noble metal nanoparticles decorated on a catalyst support with a large specific surface area can exhibit enhanced catalytic activity. To this end, a synthetic method to heterogeneously and evenly nucleate platinum nanoparticles (Pt NPs) onto mesoporous silica nanoparticles (MSNs) is developed. The obtained Pt NP-modified MSNs (Pt-MSNs) are characterized as a thin layer of 3 nm-sized Pt NPs densely assembled on the MSN surface, by which the throughput of the peroxidase-like activity of Pt-MSNs is greatly improved. The utility of Pt-MSNs in colorimetric detection of analytes is validated for two different assay schemes. Firstly, colloidally dispersed Pt-MSNs are employed as a peroxidase-mimic in a two-step cascade reaction to quantitate glucose/cholesterol based on the amount of H2O2 produced by glucose/cholesterol oxidase. Secondly, detection of C-reactive protein (CRP) is conducted on a solid substrate by adopting a sandwich immunoassay format. Detection limits are estimated to be 20 μM, 55 μM, and 3.9 pM for glucose, cholesterol, and CRP, respectively.

Mesoporous silica-supported platinum nanocatalysts for colorimetric detection of glucose, cholesterol, and C-reactive protein

Noble metal nanoparticles decorated on a catalyst support with a large specific surface area can exhibit enhanced catalytic activity. To this end, a synthetic method to heterogeneously and evenly nucleate...

Gold single atom-based aptananozyme as an ultrasensitive and selective colorimetric probe for detection of thrombin and C-reactive protein.

Anultra-efficient biocatalytic peroxidase-like Au-based single-atom nanozyme (Au-SAzymes) has been synthesizedfrom isolated Au atoms on black nitrogen doped carbon (Au-N-C) using a simple complexation-adsorption-pyrolysis method.The atomic structure of AuN4centers in black carbon was revealed bycombined high-resolution transmission electron microscopy/high-angle annular dark-field scanning transmission electron microscopy. The Au-SAzymes showed a remarkable peroxidase activity with 1.7nM as Michaelis-Menten constant, higher than most previously reported SAzyme activity. Density functional theory and Monte Carlo calculations revealed the adsorption of H2O2 on AuN4 with formation of OH* and O*. Molecular recognition was greatly enhanced via label-free integration of thiol-terminal aptamers on the surface of single Au atoms (Aptamer/Au-SAzyme) to design off-on ultrasensitive aptananozyme-based sensor for detecting thrombin and CRP with 550pM and 500pgmL-1 limits of detection, respectively. The Aptamer/Au-SAzyme showed satisfactory accuracy and precision when applied to the serum and plasma of COVID-19 patients. Due to the maximum Au atom utilization, approximately 3636 samples can be run per 1mg of gold, highlighting the commercialization potential of the developed Aptamer/Au-SAzyme approach.