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1.
A 3D ordered macroporous (3DOM) ionic‐liquid‐doped polyaniline (IL‐PANI) inverse opaline film is fabricated with an electropolymerization method and gold nanoparticles (AuNPs) are assembled on the film by electrostatic adsorption, which offers a promising basis for biomolecular immobilization due to its satisfactory chemical stability, good electronic conductivity, and excellent biocompatibility. The AuNP/IL‐PANI inverse opaline film could be used to fabricate an electrochemical impedance spectroscopy (EIS) immunosensor for the determination of Hepatitis B surface antigen (HBsAg). The concentration of HBsAg is measured using the EIS technique by monitoring the corresponding specific binding between HBsAg and HBsAb (surface antibody). The increased electron transfer resistance (Ret) values are proportional to the logarithmic value of the concentration of HBsAg. This novel immunoassay displays a linear response range between 0.032 pg mL?1 and 31.6 pg mL?1 with a detection limit of 0.001 pg mL?1. The detection of HBsAg levels in several sera showed satisfactory agreement with those using a commercial turbidimetric method.  相似文献   

2.
A streptavidin‐functionalized silver‐nanoparticle‐enriched carbon nanotube (CNT/Ag NP) is designed as trace tag for ultrasensitive multiplexed measurements of tumor markers using a disposable immunosensor array. The CNT/Ag NP nanohybrid is prepared by one‐pot in situ deposition of Ag NPs on carboxylated CNTs. The nanohybrid is functionalized with streptavidin via the inherent interaction between the protein and Ag NPs for further linkage of biotinylated signal antibodies to obtain tagged antibodies. The functionalization process greatly improves the dispersibility of the nanohybrid in water. The immunosensor array is prepared by covalently immobilizing capture antibodies on chitosan‐modified screen‐printed carbon electrodes. Through a sandwich‐type immunoreaction on the immunosensor array, numerous Ag NPs are captured onto every single immunocomplex and are further amplified by a subsequent Ag NP‐promoted deposition of silver from a silver enhancer solution to obtain the sensitive electrochemical‐stripping signal of the Ag NPs. Using carcinoembryonic antigen and α‐fetoprotein as model analytes, this proposed multiplexed immunoassay method shows acceptable precision and wide linear ranges over four orders of magnitude with detection limits down to 0.093 and 0.061 pg mL?1, respectively. The assay results of serum samples with the proposed method are in acceptable agreement with the reference values. The newly designed strategy and the functionalized tag avoid cross‐talk and the requirement of deoxygenation for electrochemical immunoassay, and thus provide a promising potential in clinical application.  相似文献   

3.
Titanium nitride (TiN), as an excellent alternative plasmonic supporting material compared to gold and silver, exhibits tunable plasmonic properties in the visible and near‐infrared spectra. However, label‐free surface plasmon resonance biosensing with TiN is seldom reported due to lack of proper surface functionalization protocols. Herein, this study reports biotinylated antibody‐functionalized TiN (BAF‐TiN) for high‐performance label‐free biosensing applications. The BAF‐TiN biosensor can quantitatively detect exosomes of 30–200 nm extracellular vesicles, isolated from a human glioma cell line. The limit of detection for an exosomal membrane protein with the BAF‐TiN biosensor is found to be 4.29 × 10?3µg mL?1 for CD63, an exosome marker, and 2.75 × 10?3µg mL?1 for epidermal growth factor receptor variant‐III, a glioma specific mutant protein, respectively. In conclusion, combining the biocompatibility, high stability, and excellent label‐free sensing performance of TiN, the BAF‐TiN biosensor could have great potential for the detection of cancer biomarkers, including exosomal surface proteins.  相似文献   

4.
A rapid microwave‐hydrothermal method has been developed to prepare monodisperse colloidal carbon nanospheres from glucose solution, and gold nanoparticles (AuNPs) are successfully assembled on the surface of the colloidal carbon nanospheres by a self‐assembly approach. The resulting AuNP/colloidal carbon nanosphere hybrid material (AuNP/C) has been characterized and is expected to offer a promising template for biomolecule immobilization and biosensor fabrication because of its satisfactory chemical stability and the good biocompatibility of AuNPs. Herein, as an example, it is demonstrated that the as‐prepared AuNP/C hybrid material can be conjugated with horseradish peroxidase‐labeled antibody (HRP‐Ab2) to fabricate HRP‐Ab2‐AuNP/C bioconjugates, which can then be used as a label for the sensitive detection of protein. The amperometric immunosensor fabricated on a carbon nanotube‐modified glass carbon electrode was very effective for antibody immobilization. The approach provided a linear response range between 0.01 and 250 ng mL?1 with a detection limit of 5.6 pg mL?1. The developed assay method was versatile, offered enhanced performances, and could be easily extended to other protein detection as well as DNA analysis.  相似文献   

5.
A novel strategy is reported for the fabrication of poly(diallyldimethylammonium chloride) (PDDA)‐protected graphene–CdSe (P‐GR‐CdSe) composites. An advanced electrogenerated chemiluminescence (ECL) immunosensor is proposed for the sensitive detection of human IgG (HIgG) by using the as‐prepared P‐GR‐CdSe composites. The P‐GR‐CdSe composite film shows high ECL intensity, good electronic conductivity, fast response, and satisfactory stability, all of which holds great promise for the fabrication of ECL biosensors with improved sensitivity. After two successive steps of amplification via the conjugation of PDDA and gold nanoparticles (GNPs) in the film, high ECL intensity is observed. The ECL immunosensor has an extremely sensitive response to HIgG in a linear range of 0.02–2000 pg mL?1 with a detection limit of 0.005 pg mL?1. The proposed sensor exhibits high specificity, good reproducibility, and long‐term stability, and may become a promising technique for protein detection.  相似文献   

6.
Current immunosensors have an insufficient number of binding sites for the recognition of biomolecules, which leads to false positive or negative results. In this research, a facile, cost‐effective, disposable, and highly selective electrochemical immunosensing platform is developed based on cationic polyelectrolyte polyallylamine (PAAMI) anchored laser‐ablated graphene (LAG). Here, for the first time, PAAMI is introduced to stabilize LAG flakes, while retaining the intrinsic thermal and electronic properties of the substrate by noncovalent π–π interaction and electrostatic physical absorption. The sensing platform offers a suitable number of anchoring sites for the immobilized antibodies by providing ? NH2 functional groups. The proper grafting of PAAMI is confirmed through X‐ray photoelectron spectroscopy and Raman spectroscopy. The immunosensing platform is applied to detect immunoglobulin (IgG) biomarkers as a proof of concept. Under optimized conditions, the sensing platform exhibits a linear range of 0.012–15 and 15–352 ng mL?1 with a limit of detection of 6 pg mL?1 for IgG detection with high selectivity. Based on the analysis, the developed immunosensing platform can be used for point‐of‐care detection of IgG in clinical diagnostic centers. Furthermore, the developed strategy is well suited for the detection of other cancer biomarkers after immobilizing the relevant antibodies.  相似文献   

7.
Surface plasmon resonance imaging (SPRi) is a unique microarray method for label‐free and multiplexed bio‐assays. However, it currently cannot be used to detect human serum samples due to its low sensitivity and poor specificity. A poly[oligo(ethylene glycol) methacrylate‐co‐glycidyl methacrylate] (POEGMA‐co‐GMA) brush was synthesized by surface‐initiated atom transfer radical polymerization (SI‐ATRP) and used as a unique supporting matrix for SPRi arrays to efficiently load probe proteins for high sensitivity while reducing nonspecific adsorptions for good selectivity. Results indicate that the polymer brush has a high protein loading capacity (1.8 protein monolayers), low non‐specific protein adsorption (below the SPR detection limit), and high immobilization stability. Three model biomarkers, α‐fetoprotein, carcinoembryonic antigen, and hepatitis B surface antigen were simultaneously detected in human serum samples by a SPRi chip for the first time, showing detection limits of 50, 20, and 100 ng mL?1, respectively. This work demonstrates great potential for a SPRi biochip as a powerful label‐free and high‐throughput detection tool in clinical diagnosis and biological research. Since the SPR detection is limited by the sensing film thickness, this approach particularly offers a unique way to significantly improve the sensitivity in the SPR detecting thickness range.  相似文献   

8.
The functionalization of a hydrogel with target molecules is one of the key steps in its various applications. Here, a versatile approach is demonstrated to functionalize a micropatterned hydrogel, which is formed by “thiol‐yne” photo‐click reaction between the yne‐ended hyperbranched poly(ether amine) (hPEA‐yne) and thiol‐containing polyhedral oligomeric silsesquioxane (PEG‐POSS‐SH). By controlling the molar ratio between hPEA‐yne and PEG‐POSS‐SH, patterned hydrogels containing thiol or yne groups are obtained. A series of thiol‐based click chemistry such as “thiol‐epoxy”, “thiol‐halogen”, “thiol‐ene”, and “thiol‐isocyanate” are used to functionalize the thiol‐containing hydrogel (Gel‐1), while the yne‐containing hydrogel (Gel‐2) is functionalized through a typical copper‐catalysed alkyne‐azide reaction (CuAAC). FTIR, UV‐vis spectra and confocal laser scanning microscopy (CLSM) are used to trace these click reactions. Due to the selective adsorption to the hydrophilic dyes, the obtained patterned hydrogel of hPEA modified with fluorescence dye is further demonstrated in application for the recognition of guest molecules.  相似文献   

9.
In immunoassays, non‐specific bindings to biosensing surfaces can be effectively prevented by formation of biocompatible and hydrophilic self‐assembled monolayer (SAM) on the surfaces. A thin gold (Au) layer on magnetic microspheres, 15 μm in diameter, enables facile SAM formation and thereby accepts second layer of filamentous virus scaffolds for the immobilization of functional proteins. The merger of the virus and SAM‐Au protected microspheres not only provides exceptionlly dense antibody loading, but also resembles biological cellular structures that enhance ligand‐receptor interactions. Site‐specific biotinylation of filamenous viruses allows formation of free‐standing virus threads (>1.0 × 1010) on streptavidin‐modified SAM‐Au microspheres. The augmented yield of antibody loading, due to the increased surface to volume ratio, on virus‐modified Au microspheres is confirmed by measuring fluorescence intensities. The bead‐based immunoassays for the detection of cardiac marker proteins exhibit increased sensitivity of virus‐Au microspheres, as low as 20 pg mL?1 of cardiac troponin I in serum, and extremely low non‐specific adsorption when compared with bare polymer beads. This increased sensitivity due to filamentous morphology and SAM‐Au layer demonstrates the feasibility of merging viruses with non‐biological materials to yield biomimetic tools for the enhanced bead‐based immunoassays.  相似文献   

10.
A novel nonenzymatic immunosensor for sensitive detection of Microcystin‐LR (MC‐LR) is constructed using a graphene platform combined with mesoporous PtRu alloy as a label for signal amplification. Primary antibody‐Microcystin‐LR (Ab1) is immobilized onto the surface of a graphene sheet (GS) through an amidation reaction between the carboxylic acid groups attached to the GS and the available amine groups of Ab1. Mesoporous PtRu alloy, prepared by corrosion PtRuAl alloys, is employed as a label to immobilize secondary antibody (Ab2). The resulting nanoparticles, PtRu‐Ab2, are used as labels for the immunosensor to detect MC‐LR. Under optimal conditions, the immunosensor exhibits a wide linear response to MC‐LR that ranges from 0.01 to 28 ng·mL?1, with a low detection limit of 9.63 pg·mL?1 MC‐LR. The proposed immunsensor shows good reproducibility, selectivity, and stability. The assayed results of polluted water with the sandwich‐type sensor are acceptable. Importantly, this methodology may provide a promising ultrasensitive assay strategy for other environmental pollutants.  相似文献   

11.
A unique strategy for effective, versatile, and facile surface biofunctionalization employing a recombinant spider silk protein genetically functionalized with the antibody‐binding Z domain (Z‐4RepCT) is reported. It is demonstrated that Z‐silk can be applied to a variety of materials and platform designs as a truly one‐step and chemical‐free surface modification that site specifically captures antibodies while simultaneously reducing nonspecific adsorption. As a model surface, SiO2 is used to optimize and characterize Z‐silk performance compared to the Z domain immobilized by a standard silanization method. First, Z‐silk adsorption is investigated and verified its biofunctionality in a long‐term stability experiment. To assess the binding capacity and protein–protein interaction stability of Z‐silk, the coating is used to capture human antibodies in various assay formats. An eightfold higher binding capacity and 40‐fold lower detection limit are obtained in the immunofluorescence assay, and the complex stability of captured antibodies is shown to be improved by a factor of 20. Applicability of Z‐silk to functionalize microfluidic devices is demonstrated by antibody detection in an electrokinetic microcapillary biosensor. To test Z‐silk for biomarker applications, real‐time detection and quantification of human immunoglobulin G are performed in a plasma sample and C1q capture from human serum using an anti‐C1q antibody.  相似文献   

12.
For biomedical application of nanoparticles, the surface chemical functionality is very important to impart additional functions, such as solubility and stability in a physiological environment, and targeting specificity as an imaging probe and a drug carrier. Although polyethylene glycol (PEG) has been used extensively, here, it is proposed that hyperbranched polyglycerol (PG) is a good or even better alternative to PEG. Superparamagnetic iron oxide nanoparticles (SPIONs) prepared using a polyol method are directly functionalized with PG through ring‐opening polymerization of glycidol. The resulting SPION‐PG is highly soluble in pure water (>40 mg mL?1) and in a phosphate buffer solution (>25 mg mL?1). Such high solubility enables separation of SPION‐PG according to size using size exclusion chromatography (SEC). The size‐separated SPION‐PG shows a gradual increase in transverse relaxivity (r2) with increasing particle size. For biological application, SPION‐PG is functionalized through multistep organic transformations (–OH → –OTs (tosylate) → –N3 → –RGD) including click chemistry as a key step to impart targeting specificity by immobilization of cyclic RGD peptide (Arg‐Gly‐Asp‐D ‐Tyr‐Lys) on the surface. The targeting effect is demonstrated by the cell experiments; SPION‐PG‐RGD is taken up by the cells overexpressing αvβ3‐integrin such as U87MG and A549.  相似文献   

13.
This study demonstrates a novel approach towards the development of advanced protein assay systems based on physically functionalized, magnetic‐core/porous‐shell CoFe2O4/SiO2 composite nanoparticles. The preparation, characterization, and measurement of the relevant properties of the protein assay system is discussed, and the system is used for the detection of cancer antigen 15‐3 (CA 15‐3, used as a model here) in clinical immunoassays. The protein assay system, based on nanometer‐sized magnetic cores and silica shells, shows good adsorption properties for the selective attachment of CA 15‐3 antibodies specific to CA 15‐3. The core/shell nanostructures exhibit good magnetic properties, which enables their integration into a quartz crystal microbalance (QCM) detection cell with the help of a permanent magnet. Under optimal conditions, the resulting immunoassay system presents a good QCM response for the detection of CA 15‐3, and allows the detection of CA 15‐3 at concentrations as low as 1.5 U mL–1 (U: units). Importantly, the proposed protein assay system can be extended to the detection of other antigens and biological compounds.  相似文献   

14.
Nanostructured porous silicon (PSi) is a promising material for the label‐free detection of biomolecules, but it currently suffers from limited applicability due to poor sensitivity, typically in micromolar range. This work presents the design, operation concept, and characterization of a novel microfluidic device and assay that integrates an oxidized PSi optical biosensor with electrokinetic focusing for a highly sensitive label‐free detection of nucleic acids. Under proper oxidation conditions, the delicate nanostructure of PSi can be preserved, while providing sufficient dielectric insulation for application of high voltages. This enables the use of signal enhancement techniques, which are based on electric fields. Here, the DNA target molecules are focused using an electric field within a finite and confined zone, and this highly concentrated analyte is delivered to an on‐chip PSi Fabry–Pérot optical transducer, prefunctionalized with capture probes. Using reflective interferometric Fourier transform spectroscopy real‐time monitoring, a 1000‐fold improvement in limit of detection is demonstrated compared to a standard assay, using the same biosensor. Thus, a measured limit of detection of 1 × 10?9 m is achieved without compromising specificity. The concepts presented herein can be readily applied to other ionic targets, paving way for the development of other highly sensitive chemical and biochemical assays.  相似文献   

15.
A low‐cost and easy‐to‐fabricate microchip remains a key challenge for the development of true point‐of‐care (POC) diagnostics. Cellulose paper and plastic are thin, light, flexible, and abundant raw materials, which make them excellent substrates for mass production of POC devices. Herein, a hybrid paper–plastic microchip (PPMC) is developed, which can be used for both single and multiplexed detection of different targets, providing flexibility in the design and fabrication of the microchip. The developed PPMC with printed electronics is evaluated for sensitive and reliable detection of a broad range of targets, such as liver and colon cancer protein biomarkers, intact Zika virus, and human papillomavirus nucleic acid amplicons. The presented approach allows a highly specific detection of the tested targets with detection limits as low as 102 ng mL?1 for protein biomarkers, 103 particle per milliliter for virus particles, and 102 copies per microliter for a target nucleic acid. This approach can potentially be considered for the development of inexpensive and stable POC microchip diagnostics and is suitable for the detection of a wide range of microbial infections and cancer biomarkers.  相似文献   

16.
Conventional lithography methods of gold patterning are based on deposition and lift‐off or deposition and etching. In this letter, we demonstrate a novel method of gold patterning using spin‐coatable gold electron‐beam resist which is functionalized gold nanocrystals with amine ligands. Amine‐stabilized gold electron beam resist exhibits good sensitivity, 3.0 mC/cm2, compared to that of thiol‐stabilized gold electron beam resists. The proposed method reduces the number of processing steps and provides greater freedom in the patterning of complex nanostructures.  相似文献   

17.
A real‐time, sensitive, and selective detection device to monitor the healing status of chronic wounds at the point of care is urgently required to render the management of this disease more effective. The photonic properties of porous silicon resonant microcavity (pSiRM) afford an excellent opportunity to be developed as a highly sensitive optical biosensor to monitor the presence of specific biomarkers found in the wound exudate, such as matrix metalloproteinases (MMPs). In this study, the pSiRM is designed, fabricated, and functionalized using a fluorogenic MMP peptide substrate featuring both a fluorophore and a quencher. The peptide‐functionalized pSiRM is then employed as a fluorescence‐based optical biosensor for MMPs. Active MMPs recognize and cleave the peptide sequence of the substrate, producing an immobilized peptide fragment carrying the fluorophore. The fluorescence intensity of the fluorophore embedded within the pSiRM matrix is enhanced by the photonic structure of the pSiRM compared to other pSi photonic structures. This fluorescence enhancement translates into high sensitivity, enabling detection of MMP‐1 at a limit of detection as low as 7.5 × 10?19 m after only 15 min incubation time. Finally, the biosensor also allows the detection and quantification of the concentration of MMPs in human wound fluid.  相似文献   

18.
Highly sensitive and selective chemiresistive sensors based on graphene functionalized by metals and metal oxides have attracted considerable attention in the fields of environmental monitoring and medical assessment because of their ultrasensitive gas detecting performance and cost‐effective fabrication. However, their operation, in terms of detection limit and reliability, is limited in traditional applications because of ambient humidity. Here, the enhanced sensitivity and selectivity of single‐stranded DNA‐functionalized graphene (ssDNA‐FG) sensors to NH3 and H2S vapors at high humidity are demonstrated and their sensing mechanism is suggested. It is found that depositing a layer of ssDNA molecules leads to effective modulation of carrier density in graphene, as a negative‐potential gating agent and the formation of an additional ion conduction path for proton hopping in the layer of hydronium ions (H3O+) at high humidity (>80%). Considering that selectively responsive chemical vapors are biomarkers associated with human diseases, the obtained results strongly suggest that ssDNA‐FG sensors can be the key to developing a high‐performance exhaled breath analyzer for diagnosing halitosis and kidney disorder.  相似文献   

19.
Circulating tumor cells (CTCs) play key roles in the development of tumor metastasis. It remains a significant challenge to capture and detect CTCs with high purity and sensitivity from blood samples. Herein, a nanoplatform is developed for the efficient isolation and ultrasensitive detection of CTCs by combining near‐infrared (NIR) multivalent aptamer functionalized Ag2S nanodots with hybrid cell membrane‐coated magnetic nanoparticles. Multivalent aptamer functionalized Ag2S nanodots are synthesized using a one‐pot method under mild conditions (60 °C). White blood cell and tumor cell membranes are fused as the hybrid membrane and coated with magnetic nanoparticles, which are further modified with streptavidin (SA). Through the specific interaction of SA‐biotin, the multivalent aptamer‐Ag2S nanodots are grafted with hybrid cell membrane‐magnetic nanoparticles. Due to the features of hybrid cell membrane modification, multivalent aptamer functionalization, magnetic separation, and NIR fluorescence measurements, the nanoplatform shows sensitive recognition, efficient capture, easy isolation, and sensitive detection of CTCs due to its great enhancement in anti‐interference from background and improvement on binding ability toward CTCs. The capture efficiency and purity for CTCs is as high as 97.63% and 96.96%, respectively. Furthermore, the nanoplatform is successfully applied to the detection of CTCs in blood samples.  相似文献   

20.
Mesoporous CoNi@Au core@shell nanorods are synthesized as magnetic drug nanocarriers by electrodeposition using ionic liquid‐in‐aqueous microemulsions. Mesoporous nanorods present a highly effective area (186 m2 g?1) and magnetic character that allows their manipulation, concentration, and retention by applying a magnetic field. The nanorods have been functionalized with thiol‐poly(ethyleneglycol) molecules, and molecules of Irinotecan, a drug used in chemotherapy, are retained in both the lattice of the linked thiol‐poly(ethyleneglycol) molecules and inside the interconnected nanorods pores. The nanorods' mesoporous character allows a high drug‐loading capability and magnetic behavior that allows the drug's controlled release. A high cellular viability of HeLa cells is obtained after their incubation with the nanorods functionalized with thiol‐poly(ethyleneglycol). However, when the nanorods function as carriers for Irinotecan, significant cell death is occurred when HeLa cells are incubated with the functionalized, drug‐loaded nanorods. Cell death is also produced by applying an alternating magnetic field due to both the effect of the release of Irinotecan from the carrier as to mechanical damage of cells by nanorods subjected to the effect of a magnetic field. The proposal to used mesoporous magnetic nanorods as drug carriers can thus dramatically reduce the amounts of both nanocarrier and drug needed to efficiently destroy cancer cells.  相似文献   

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