Gold Nanoflowers Research Articles

A durable hydrophobic photothermal membrane based on a honeycomb structure MXene for stable and efficient solar desalination.

Solar powered water evaporation is a green and environmentally friendly water treatment technology, which is a hot research topic for water purification at present. Advanced structural design and hydrophilic photothermal materials have achieved efficient solar evaporation of pure water, but the long-term stability of high salinity desalination has become a problem that cannot be ignored in practical applications. In order to solve this problem, a hydrophobic honeycomb structure MXene/AuNFs composite membrane was proposed in this paper, which used the three-dimensional highly porous microstructure of MXene and multibranched structure of gold nanoflowers particles to improve the light absorption and photothermal conversion efficiency of MXene/AuNFs. At the same time, the surface of the composite membrane was modified with hydrophobic fluorosilane 1H,1H,2H,2H-perfluorodecyltriethoxysilane (PFTE). The hydrophobic layer can prevent the accumulation of salt particles on the surface of the membrane, so that the composite film can continue to produce water vapor in a high salt environment. With high utilization rate of light energy, multiple-level geometrical structures of MXene for rapid water transport on the filter membrane and salt barrier on the membrane good stability, the hydrophobic MXene/AuNFs achieves solar evaporation rate of 1.59 kg m-2 h-1 and solar conversion efficiency is 97.8%, and stable operation under simulated sea water conditions under one sun irradiation over more than 10 cycles. The hydrophobic MXene/AuNFs membrane proved to be an efficient and stable photothermal material for solar desalination.

Controllable synthesis of multi-tip spatial gold nanostructures to facilitate SPR enhancement for exosomal PD-L1 assay

A surface plasmon resonance (SPR) biosensor was developed utilizing gold nanoflowers with multi-tip tiny petals (Au@Au mNFs) and multifunctional peptides for the selective detection of exosomal programmed cell death 1 ligand 1 (ExoPD-L1). In the presence of different salt concentrations, DNA-functionalized gold seeds underwent in situ gold growth, resulting in Au@Au NFs with varying numbers of petals or Au@Au nanospheres (Au@Au NSs). The growth orientation of the gold shells was influenced by the “upright”, “tilted” or “lying” state of the DNA on the gold core surface, depending on the salt concentration solution. The finally prepared Au@Au mNFs exhibited a multi-tip spatial structure, which served as a highly efficient SPR sensitized layer with abundant hot spots on the gold film surface. The enhanced SPR sensitivity was attributed to the substantial electromagnetic field enhancement generated at the tips of the mNFs. By achieving suitable surface coverage of the multifunctional peptide at the sensing interface constructed by Au@Au mNFs, efficient capture of ExoPD-L1 was ensured. The analysis results demonstrated that the developed SPR sensor successfully detected ExoPD-L1 within a concentration range of 10–107 particles/mL, with a detection limit of 4.95 particles/mL. This work highlights the significant improvement in SPR sensing and detection performance achieved by utilizing Au@Au mNFs with multi-tip spatial nanostructures, providing a straightforward approach for achieving ultra-sensitive detection of target substances through SPR.

Planting gold nanoflower for harvesting reproducible SERS substrate

Surface-enhanced Raman scattering (SERS) is an ultrasensitive analytical method which has been applied in many fields, and the reproducibility of the substrate is important for reliable SERS analysis. In present work, an innovative method inspired by the flower planting process is put forward to acquire gold nanoflower (AuNF) SERS substrate. Three steps (digging holes, sowing the gold nanoseeds and seeds grow into gold nanoflowers) are included in the substrate fabrication process, and the influence of preparing conditions (like reacting time and Na3Au(SO3)2 concentration) on the substrate morphology and SERS performance are investigated. The acquired AuNF substrate not only exhibits good SERS performance but also possesses excellent reproducibility while being used to detect the rhodamine 6G (R6G) molecular. The relative standard deviation (RSD) of Raman signals among substrates acquired in distinct batches (substrate-to-substrate) is as low as 6.67 %. Since the AuNF substrate is prepared by the wet chemistry route based on seed-mediated growth and there are no expensive reagents or complicated process used, the new process to obtain AuNF substrate is cost-effective and easy to scale up.

A sensitive signal off photoelectrochemical aptasensor based on CuO nanosheets @Au nanoflowers for the detection of chloramphenicol

A signal off type photoelectrochemical (PEC) aptasensor was constructed for the sensitive detection of chloramphenicol based on CuO nanosheets (CuO NSs) and gold nanoflowers (Au NFs) modified indium tin oxide (ITO) electrode. CuO NSs was p-type photoactive semiconductor materials, which had narrow band gap and high absorption rate of visible light. After combining with Au NFs, the composite CuO NSs@Au NFs produced a strong cathodic photocurrent for the excellent electron conductivity, large specific surface area and Surface Plasmon Resonance effect (SPR) of Au NFs. At the same time, Au NFs provided a binding site for aptamer, which greatly improved the sensitivity of the constructed sensing platform. After the target chloramphenicol (CAP) was specifically captured by the aptamers on the electrode, the formed bioconjugates seriously hindered the transfer of electrons and the absorption of light and caused a reduced photocurrent. For the detection of CAP, this signal off type PEC aptasensor had a linear range of 5 pM-300 nM and a limit of detection of 49.9 fM (S/N = 3) with good stability, selectivity and reproducibility. Meanwhile, the method had satisfactory recovery results in the detection of CAP in real-life samples such as milk, tap water, honey and chloramphenicol eye drops.

Pt@AuNF nanozyme and horseradish peroxidase-based lateral flow immunoassay dual enzymes signal amplification strategy for sensitive detection of zearalenone

Lateral flow immunoassay (LFIA) has been employed extensively for the rapid, accurate, and portable detection of foodborne toxins. Here, the platinum gold nanoflower core-shell (Pt@AuNF) nanozyme with excellent optical properties, good catalytic ability and controllable reaction conditions were prepared to effectively improve the performance of lateral flow immunoassay (LFIA) strips. The Pt@AuNF nanozyme and horseradish peroxidase (HRP) combined with monoclonal antibody were used as signal probes based on the dual enzymes catalytic signal amplification strategy to detect Zearalenone sensitively. Dual enzymes catalyze the decomposition of hydrogen peroxide into hydroxyl radicals, and under the influence of hydroxyl radicals, colorless 3,3′,5,5′ -tetramethylbenzidine (TMB) is oxidized to blue ox-TMB, which is superimposed on the strips for signal amplification to broaden the detection range. The limit of detection (LOD) of the Pt@AuNF-HRP labeled LFIA strips after signal amplification was 0.052 ng/mL, and the detection range was 0.052–7.21 ng/mL. Compared with the Pt@AuNF labeled strips, while reducing the probes amount by half to achieve antibody conservation, the detection range was expanded by 5-fold based on achieving improved sensitivity. The study provided a meaningful reference for expanding the detection range based on immunoassay.

Development of a Rapid Gold Nanoflowers Immunochromatographic Test Strip Based on the Nanobody for Detection of Aflatoxin B1

AbstractA rapid gold nanoflowers (AuNFs) immunochromatographic test strip (ICTS) based on the nanobody (Nb) was developed and used for rapid detection of Aflatoxin B1 (AFB1). Thanks to remarkable physicochemical stability, structural adaptability, ease of genetic manipulation, expression in the prokaryotic system with high yield and tolerance to harsh environments of nanobodies, the G8‐DIG was produced via expression and purification, which was labeled with AuNFs by electrostatic adsorption as signal probes in the ICTS. The AuNFs‐anti‐G8‐DIG nanobodies as a probe of ICTS could specifically recognize not only AFB1 but also DIG‐BSA. In this study, the DIG‐BSA replaced the anti‐6×His‐tag antibody of traditional nanobody‐based ICTS was coated on the control line, which was easier to synthesize, cheaper, and more stable. Under the optimal conditions, the AuNFs‐based nanobody ICTS system accurately detected AFB1 linearly and dynamically over the range (IC20~IC80) of 1.02~27.86 ng/mL, with the IC10 and IC50 of 0.1 ng/mL and 5.46 ng/mL respectively. In addition, these developed nanobodies ICTS had a universal application in the analysis of AFB1 in corn samples.

Machine learning-powered electrochemical aptasensor for simultaneous monitoring of di(2-ethylhexyl) phthalate and bisphenol A in variable pH environments

Plastic waste is a pernicious environmental pollutant that threatens ecosystems and human health by releasing contaminants including di(2-ethylhexyl) phthalate (DEHP) and bisphenol A (BPA). Therefore, a machine-learning (ML)-powered electrochemical aptasensor was developed in this study for simultaneously detecting DEHP and BPA in river waters, particularly to minimize the electrochemical signal errors caused by varying pH levels. The aptasensor leverages a straightforward and effective surface modification strategy featuring gold nanoflowers to achieve low detection limits for DEHP and BPA (0.58 and 0.59 pg/mL, respectively), excellent specificity, and stability. The least-squares boosting (LSBoost) algorithm was introduced to reliably monitor the targets regardless of pH; it employs a layer that adjusts the number of multi-indexes and the parallel learning structure of an ensemble model to accurately predict concentrations by preventing overfitting and enhancing the learning effect. The ML-powered aptasensor successfully detected targets in 12 river sites with diverse pH values, exhibiting higher accuracy and reliability. To our knowledge, the platform proposed in this study is the first attempt to utilize ML for the simultaneous assessment of DEHP and BPA. This breakthrough allows for comprehensive investigations into the effects of contamination originating from diverse plastics by eliminating external interferent-caused influences.

Facile synthesis of gold nanoflowers and their application in glycerol electro-oxidation

Gold nanoflowers were prepared using the Satureja macrostema aqueous extract. The size of the nanoflowers was determined to be an average of 730 nm in length, with a thickness of 10 nm, as measured by scanning electron microscopy (SEM) and DLS. Transmission electron microscopy (TEM) and X-ray diffraction (XRD) confirmed the nanoscale size and the FCC cubic crystalline structure of the flowers, with a crystallite size of 10 nm, and the specific surface area using the BET method was 36.53 m2g−1. Infrared (FTIR) and gas chromatography coupled to mass spectrometry (GC–MS) were used to identify the reducing compounds responsible for forming the nanoflowers from HAuCl4. The nanoflowers exhibited a maximum current density of 1510 mA mg−1 and 819 mA mg−1 for the electro-oxidation of analytical and crude glycerol in an alkaline medium, respectively. The nanoflowers were found to be stable after 60 min of chronoamperometry testing, which may be attributed to their morphology, preferential orientation, and the remaining surface molecules of the plant extract.

Resveratrol-Coated Gold Nanoflowers for CT Imaging and Apoptosis/Photothermal Synergistic Therapy of Malignant Melanoma.

In the past decade, photothermal therapy (PTT) of tumors based on gold nanomaterials has been widely studied because of their strong extinction ability and high photothermal conversion ability in the near-infrared (NIR) region. However, related research still faces two problems: First, the biosafety of the surface ligands on gold nanomaterials is not ideal and even has strong toxicity, so the surface modification or shell coating is very necessary; second, gold nanomaterials only have a single PTT function, which requires high temperature to achieve better treatment effect. Therefore, it is necessary to enrich the antitumor function of gold nanomaterials and realize synergistic therapy. Natural polyphenols can combine with each other or other substances through various supramolecular forces, forming shells on the surface of nanomaterials and reducing biotoxicity. In addition, natural polyphenols represented by resveratrol have antitumor activity and can induce apoptosis of tumor cells. Therefore, the surface coating method of gold nanomaterials with natural polyphenols with antitumor activity can effectively solve the above problems. In this work, we prepared resveratrol-coated gold nanoflowers (Au@Res NFs) and applied them to the treatment of malignant melanoma. Resveratrol in Au@Res NFs can induce the apoptosis of tumor cells, and Au@Res NFs can play a role in PTT under an NIR laser. In cell experiments, the synergistic effect of apoptosis/PTT on the A375 cells was extremely strong. In animal experiments, Au@Res NFs enriched in tumor sites identified the location and boundary of tumors by computed tomography (CT). The apoptosis induced by resveratrol had a certain inhibitory effect on tumor growth. Further applying the NIR laser, under the synergistic effect of apoptosis and PTT, the tumors were completely eliminated without recurrence during the experimental period.

Aptamer-based carbohydrate antigen 125 sensor with molybdenum disulfide functional hybrid materials

Epithelial ovarian cancer is a malignant tumor of the female reproductive system with insidious symptoms, aggressiveness, risk of metastasis, and high mortality. Carbohydrate antigen 125 (CA125), a standard biomarker for screening epithelial ovarian cancer, can be applied to track cancer progression and treatment response. Here, we constructed an aptamer-based electrochemical biosensor to achieve sensitive detection of CA125. Molybdenum disulfide (MoS2) was used as the stable layered substrate, combined with the irregular branched structure of gold nanoflowers (AuNFs) to provide the sensing interface with a large specific surface area by one-step electrodeposition AuNFs@MoS2. The simplified electrode modification step increased the stability of the electrode while ensuring excellent electrochemical performance and providing many sulfhydryl binding sites. Then, AuNFs@MoS2/CA125 aptamer/MCH sensor was designed for CA125 detection. Based on AuNFs@MoS2 electrode, CA125 aptamer with sulfhydryl as the sensitive layer was fixed on the electrode by gold sulfur bonds. 6-Mercapto-1-hexanol (MCH) was used to block the electrode and reduce the non-specific adsorption. Finally, DPV analysis was applied for CA125 detection with the range of 0.0001 U/mL to 500 U/mL. Our designed aptamer sensor showed reasonable specificity, reproducibility, and stability. Clinical sample testing also proved the consistency of our sensor with the gold standard in negative/positive judgment. This work demonstrated a novel strategy for integrating nanostructures and biocompatibility to build advanced cancer biomarker sensors with promising applications.

A gold nanoflower based dual mode aptasensor for aflatoxin B1 detection using SERS and fluorescence effect simultaneously

Aflatoxin B1 (AFB1) is usually the major aflatoxin produced by toxigenic strains and has been identified the most potent natural carcinogen. Here, a SERS/fluorescence dual-mode nanosensor has been designed while gold nanoflowers (AuNFs) was used as substrate for the detection of AFB1. AuNFs exhibited excellent SERS enhancement effect as well as the good fluorescence quenching effect which made the dual signal detection possible. First, the surface of AuNFs was modified with AFB1 aptamer via Au-SH group. Then, the complementary sequence functionalized with Cy5 (the signal molecule) was attached to AuNFs based on the base complementary pairing principle. On this case, Cy5 was close to AuNFs, the SERS intensity was greatly enhanced and the fluorescence intensity was quenched. After incubation with AFB1, the aptamer was preferentially combined to its target AFB1. Thus, the complementary sequence detached from AuNFs which caused the SERS intensity of Cy5 decreased while its fluorescence effect recovered. Then, the quantitative detection was realized with two optical properties. The LOD was calculated to be 0.03 ng/mL. It was a convenient and fast detection method which expanded the application of nanomaterials based multi-signal simultaneous detection.

Stable gold nanoflowers modified by zwitterionic sulfhydryl sulfobetaine for enhanced antitumor efficiency

Gold nanoflowers are flower-like gold nanoparticles with high photothermal conversion efficiency. Unfortunately, many gold nanoflowers have complicated synthesis methods using toxicity molecules and lack colloidal stability. In this work, in order to improve the long-term colloidal stability of existing gold nanoflowers, sulfhydryl sulfobetaine (SB-SH) was modified on the surface of gold nanoflowers by S-Au bonds to produce SB-SH stabilized gold nanoflowers (SB-SH@AuNFs). SB-SH@AuNFs did not generate any precipitation within 24 h and had a high photothermal conversion efficiency at 38.9%. In addition, the relative cell viability of A549 cells was only 11% with 0.193 mM SB-SH@AuNFs under laser irradiation. More importantly, SB-SH@AuNFs had the best antitumor effect in vivo under 808 nm laser irradiation. Thus, stable SB-SH@AuNFs with high photothermal conversion efficient were synthesized by using zwitterionic molecules. The modification of gold nanoflowers using zwitterionic molecules here is a new method to improve the antitumor efficiency of noble metal photothermal agents.

Sensitive and rapid detection of tetrodotoxin based on gold nanoflower-and latex microsphere-labeled monoclonal antibodies.

Tetrodotoxin (TTX) could result in serious diseases due to its extremely high neurotoxicity. Thus, it is of great importance to measure TTX for food safety. In this study, an anti-TTX monoclonal antibody with good specificity and high affinity was used to develop the immunochromatographic test strips (ICTS). Gold nanoflower (AuNF) with multiple branches and latex microsphere (LM) with large particle size as signal reporters were employed for improving the sensitivity of test strips. Both AuNF and LM probes are stable, and the developed ICTS were specific to TTX, demonstrating no cross-reactivity with other marine toxins. The linear range of AuNF- and LM-based strips for TTX was 9.49-330.98ng/mL and 5.40-443.19ng/mL, respectively. The limit of detection (LOD) of AuNF- and LM-based strips was determined to be 9.49ng/mL and 5.40ng/mL, respectively. In summary, the developed ICTS based on AuNF and LM signal probes displayed enhancement of sensitivity and provided rapid and specific detection of TTX.

Two-color lateral flow nucleic acid assay combined with double-tailed recombinase polymerase amplification for simultaneous detection of chicken and duck adulteration in mutton

We propose a visual strategy for simultaneous detection of chicken and duck ingredients in mutton by integration of a double tailed multiplex recombinase polymerase amplification (mRPA) assay with a multicolor nucleic acid hybridization lateral flow strip (mNAH-LFS) assay. Two colored gold nanoparticles (AuNPs), including gold nanospheres (red) and gold nanoflowers (blue), were synthesized and functionalized with the corresponding nucleic acid probe to target each analyte. The RPA product was analyzed by this two-colored mNAH-LFS assay which includes a red T1 line for chicken adulteration and a blue T2 line for duck adulteration. In this RPA reaction, special Spacer9 and tail nucleotide sequence-tagged primers (Tag+Spacer9+F-primer, Tag+Spacer9+R-primer) were used to obtain double-stranded amplicons with specific wobbled sequences that can directly bind with AuNPs-labeled probes (AuNSs-cT1 hybridized with T1, AuNFs-cT4 hybridized with T4) and solid-phase probes (biotin-cT2 hybridized with T2, biotin-cT3 hybridized with T3) on the mNAH-LFS producing a red or/and cyan color on the T lines. The detection limits of this mRPA-mNAH-LFS assay for chicken and duck were 0.01% and 0.01%, respectively. Finally, this mRPA-mNAH-LFS assay was successfully used to authenticate 59 commercial meat products. In conclusion, this mRPA-mNAH-LFS is a sensitive, accurate, and cost-effective approach for meat quality assurance and traceability.

Zebrafish (Danio rerio) TRβ- and TTR-based electrochemical biosensors: Construction and application for the evaluation of thyroid-disrupting activity of bisphenols

Thyroid-disrupting chemicals (TDCs) have received increasing concerns because of their negative health impacts on both wildlife and humans. This study aimed to develop in vitro screening assays for TDCs based on thyroid hormone receptor β (TRβ) and transthyretin (TTR) proteins. Firstly, the recombinant ligand-binding domain of TRβ (TRβ-LBD) and TTR proteins of zebrafish were produced by eukaryotic expression system and then used as bio-recognition components to construct electrochemical biosensors. In the biosensors, the supported bilayer lipid membrane (s-BLM) was used as a matrix to immobilize proteins, and gold nanoflowers (AuNFs) were used to improve the sensitivity by increasing electroactive surface area. Under the optimizing conditions, the zfTRβ-LBD/AuNFs/s-BLM/GCE biosensor had a detection range of 0.23 nM–1.92 μM and a detection limit of 0.07 nM for triiodothyronine (T3), while the zfTTR/AuNFs/s-BLM/GCE biosensor had a detection range of 0.46 nM–3.84 μM, with a detection limit of 0.13 nM. Based on the constructed biosensors, the order of T3 equivalent concentrations of bisphenols was BPA ≈ BPS > BPF > BPAF ≈ BPAP > BPZ, which was similar to the results of recombinant TRβ two-hybrid yeast assay. Furthermore, the reliability of the biosensors was validated by molecular docking, in which BPA and BPS showed higher binding affinity to zfTRβ-LBD. Therefore, this study provided a valuable tool for efficiently screening TDCs.

A novel label-free graphene oxide nano-wall surface decorated with gold nano-flower biosensor for electrochemical detection of brucellosis antibodies in human serum

Conventional serological methods to detect brucellosis are time-consuming, expensive, and require special equipment, but they are less sensitive than newly developed approaches. To overcome these drawbacks, in addition to the ultrasensitive, rapid, easy, and early detection of brucellosis biomarkers, more emphasis on novel methods through electrochemical nanostructured platforms is needed. A novel nanostructured platform made of graphene oxide (GO) nanosheet and gold nanoparticles (AuNPs), with field emission scanning electron microscopy (FESEM) scanning revealing that the GO is vertically arranged on the surface like nano-walls (GONW) and the AuNPs are decorated in the shape of nanoflowers. FESEM, energy dispersive spectroscopy (EDS), and Fourier transform infrared (FTIR) were used to characterize the successfully fabricated nanostructured surface. The electrochemical impedance spectroscopy (EIS), differential pulse voltammetry (DPV), and cyclic voltammetry (CV) techniques applied to the fabricated graphene oxide nanowall-gold nanoflower (GONW@Au-NF) platform indicated excellent electroactivity (more than a 40% increase) and valuable antifouling properties, which make it more sensitive than bare fluorine-doped tin oxide (FTO) electrodes. The limit of detection (LOD) using different concentrations of Brucella–against antibody (Br-Ab) is estimated at 9.3 fg mL−1. The selectivity of the biosensor was tested using E.coli antibody and bovine serum albumin (BSA). The results revealed good selectivity (significant difference) and acceptable reproducibility (not exceeding 12% RSD). Rapid and early detection of Br-Ab in both PBS and real samples (serum), with excellent sensitivity and selectivity, proposed our fabricated biosensor as an ideal candidate for brucellosis biomarker detection.

Ultrasensitive colorimetric detection of tetracyclines based on in-situ growth of gold nanoflowers.

A colorimetric method based on in-situ generation of gold nanoflowers for the detection of tetracyclines (TCs) was proposed. We found that gold nanoflowers could be formed in the HAuCl4-NH2OH redox reaction directly without the addition of small-sized gold nanoparticles (Au NPs) as seeds when an alkaline borax buffer solution was employed as the reaction medium. Interestingly, the shape and size of the generated gold nanoflowers were regulated with TC. Briefly, large flower-like gold nanoparticles were formed with a low concentration of TC while small spherical gold nanoparticles were generated with a high concentration of TC. The generated gold nanoflowers exhibited different surface plasmon absorption (SPR) properties. Thus, a simple and rapid colorimetric method was established for the detection of TC antibiotics. This method exhibited high sensitivity for the detection of TC, oxytetracycline (OTC), and doxycycline (DC) with detection limits of 2.23nM, 1.19nM, and 5.81nM, respectively. The proposed colorimetric method was applied to the determination of TC in both milk samples and water samples.

Detection of plasma miR-223 by a novel label-free graphene oxide/gold nanocomposite immunosensor in colorectal cancer patients: An electrochemical biosensor approach

MicroRNAs (miRNAs) are one group of the pertinent molecular biomarkers in the early detection of cancers, especially colorectal cancer. A simple and label-free nanostructured electrochemical biosensor was fabricated to detect miR-223, a colorectal cancer miRNA, by a vertical arrangement of graphene oxide (GO) decorated with gold (Au) Nano-flower nanostructures (GO@Au-NS). The use of high surface-to-volume aspect ratio nanostructures with excellent electroactivity that contain thiolated DNA probes (Cap-223) could increase the accessibility, chance, and recognition performance of miR-223. The substrate characteristics are scanned by HRSEM, EDX, and electrochemical techniques. Furthermore, the successful immobilization of Cap-223 on GO@Au-NS and their hybridization with targeted miR-223 macro-molecules were investigated by electrochemical techniques (CV, EIS, and DPV). The electroactivity of the working electrode before and after GO@Au-NS fabrication showed a significant difference. A wide range of linear detection of miR-223 by a fabricated biosensor was from zM to nM, with a lower detection limit (LOD) of 0.012 aM. The selectivity of the nanostructured miR-223 biosensor was also investigated by studying the discrimination ability of mismatched miRNAs sequences, which resulted in high selectivity for the biosensor. In addition, its efficiency in the detection of miR-223 in a real sample (human serum) showed a remarkable ability of the biosensor to detect it with a low percent of relative standard deviation (RSD = 5.7%). The developed electrochemical biosensor exhibited great potential for miRNA-based early detection of colorectal cancer and had excellent sensitivity, and selectivity, making it helpful for use in translational medicine research and clinical applications.

Duplex Electrochemical Microfluidic Sensor for COVID-19 Antibody Detection: Natural versus Vaccine-Induced Humoral Response.

The rapid transmission and resilience of coronavirus disease 2019 (COVID-19) have led to urgent demands in monitoring humoral response for effective vaccine development, thus a multiplex co-detection platform to discriminate infection-induced from vaccine-induced antibodies is needed. Here a duplex electrochemical immunosensor for co-detection of anti-nucleocapsid IgG (N-IgG) and anti-spike IgG (S-IgG) is developed by using a two-working electrode system, via an indirect immunoassay, with antibody quantification obtained by differential pulse voltammetry. The screen-printed electrodes (SPEs) are modified by carbon black and electrodeposited gold nanoflowers for maximized surface areas, enabling the construction of an immunological chain for S-IgG and N-IgG electrochemical detection with enhanced performance. Using an optimized immunoassay protocol, a wide linear range between 30-750 and 20-1000ngmL-1 , and a limit of detection of 28 and 15ngmL-1 are achieved to detect N-IgG and S-IgG simultaneously in serum samples. This duplex immunosensor is then integrated in a microfluidic device to obtain significantly reduced detection time (≤ 7min) while maintaining its analytical performance. The duplex microfluidic immunosensor can be easily expanded into multiplex format to achieve high throughput screening for the sero-surveillance of COVID-19 and other infectiousdiseases.

Ultrasensitive amperometric determination of hand, foot and mouth disease based on gold nanoflower modified microelectrode

Given the widespread use of point-of-care testing for diagnosis of disease, micro-scale electrochemical deoxyribonucleic acid (DNA) biosensors have become a promising area of research owing to its fast mass transfer, high current density and rapid response. In this study, a gold nanoparticles modified gold microelectrode (AuNPs/Au–Me) was constructed to determine the hand, foot and mouth disease (HFMD)-related gene. The noble metal nanoparticles modification yielded ca. 7.4-fold increase in electroactive surface area of microelectrode, and the signal for HFMD-related gene was largely magnified. Under optimal conditions, the biosensor exhibited salient selectivity and sensitivity with a low detection limit of 0.3 fM (S/N = 3), which is sufficient for clinical diagnosis of HFMD. Additionally, the developed AuNPs/Au–Me was successfully applied to determining the polymerase chain reaction (PCR) amplified products of target gene. Thus, the electrochemical DNA biosensor possesses great potential in early-stage diagnosis and long-term monitoring of various disease.