Assembly of Multifunctionalized Gold Nanoparticles with Chemiluminescent, Catalytic, and Immune Activity for Label-Free Immunoassays

In this study, we report a universal label-free immunoassay to detect antigen based on multifunctionalized gold nanoparticles (MF-GNPs), which were obtained by successive assembly of <i>N</i>-aminobutyl-<i>N</i>-ethylisoluminol functionalized gold nanoparticles (ABEI-GNPs) with antibody, bovine serum albumin (BSA) and Co²⁺. MF-GNPs exhibited excellent chemiluminescent (CL), catalytic and immune activity. It was demonstrated that the CL signal of MF-GNPs decreased in the presence of antigens via antigen–antibody specific binding using human immunoglobulin G (hIgG) and corresponding antibody goat anti-human IgG (anti-hIgG) as a model system, due to that immunoreaction led to the aggregation of GNPs. According to the decreased CL intensity, hIgG could be determined in the range of 1.0 fM to 1.0 nM with a low detection limit of 0.13 fM. Furthermore, this CL strategy was also confirmed to be a general one by replacing hIgG with heart-type fatty acid-binding protein (H-FABP), which is a biomarker of early acute myocardial infarction (AMI). The CL strategy could be employed to detect H-FABP ranging from 10.0 fM to 10.0 nM, and the detection limit is 7.8 fM. The CL strategy also showed good selectivity. It might be extended to detect other antigens if their corresponding antibodies are available

Highly Chemiluminescent Magnetic Beads for Label-Free Sensing of 2,4,6-Trinitrotoluene

Until now, despite the great success acquired in scientific research and commercial applications, magnetic beads (MBs) have been used for nothing more than a carrier in most cases in bioassays. In this work, highly chemiluminescent magnetic beads containing <i>N</i>-(4-aminobutyl)-<i>N</i>-ethyl isoluminol (ABEI) and Co²⁺ (Co²⁺/ABEI/MBs) were first synthesized via a facile strategy. ABEI and Co²⁺ were grafted onto the surface of carboxylated MBs by virtue of a carboxyl group and electrostatic interaction. The as-prepared Co²⁺/ABEI/MBs exhibited good paramagnetic properties, satisfactory stability, and intense chemiluminescence (CL) emission when reacted with H₂O₂, which was more than 150 times that of ABEI functionalized MBs. Furthermore, it was found that 2,4,6-trinitrotoluene (TNT) aptamer could attach to the surface of Co²⁺/ABEI/MBs via electrostatic interaction and coordination interaction between TNT aptamer and Co²⁺, leading to a decrease in CL intensity due to the catalytic site Co²⁺ being blocked by the aptamer. In the presence of TNT, TNT would bind strongly with TNT aptamer and detach from the surface of Co²⁺/ABEI/MBs, resulting in partial restoration of the CL signal. Accordingly, label-free aptasensor was developed for the determination of TNT in the range of 0.05–25 ng/mL with a detection limit of 17 pg/mL. This work demonstrates that Co²⁺/ABEI/MBs are easily connected with recognition biomolecules, which are not only magnetic carriers but also direct sensing interfaces with excellent CL activity. It provides a novel CL interface with a magnetic property which easily separates analytes from the sample matrix to construct label-free bioassays

Dynamically Tunable Chemiluminescence of Luminol-Functionalized Silver Nanoparticles and Its Application to Protein Sensing Arrays

It is still a great challenge to develop an array-based sensing system that can obtain only multiparameters, according to a single experiment and device. The role of conventional chemiluminescence (CL) in biosensing has been limited to a signal transducer in which a single signal (CL intensity) can be obtained for quantifying the concentrations of analytes. In this work, we have developed an dynamically tunable CL system, based on the reaction of luminol-functionalized silver nanoparticles (luminol–AgNPs) with H₂O₂, which could be tunable via adjusting various conditions such as the concentration of H₂O₂, pH value, and addition of protein. A single experiment operation could obtain multiparameters including CL intensity, the time to appear CL emission and the time to reach CL peak value. The tunable, low-background, and highly reproducible CL system based on luminol–AgNPs is applied, for the first time, as a sensing platform with trichannel properties for protein sensing arrays by principal component analysis. Identification of 35 unknowns demonstrated a success rate of >96%. The developed sensing arrays based on the luminol–AgNPs provide a new way to use nanoparticles-based CL for the fabrication of sensing arrays and hold great promise for biomedical application in the future

Highly Chemiluminescent Graphene Oxide Hybrids Bifunctionalized by <i>N</i>‑(Aminobutyl)‑<i>N</i>‑(Ethylisoluminol)/Horseradish Peroxidase and Sensitive Sensing of Hydrogen Peroxide

<i>N</i>-aminobutyl-<i>N</i>-ethylisoluminol and horseradish peroxidase bifunctionalized graphene oxide hybrids (ABEI-GO@HRP) were prepared through a facile and green strategy for the first time. The hybrids exhibited excellent chemiluminescence (CL) activity over a wide range of pH from 6.1 to 13.0 when reacted with H₂O₂, whereas ABEI functionalized GO had no CL emission at neutral pH and showed more than 2 orders of magnitude lower CL intensity than ABEI-GO@HRP at pH 13.0. Such strong CL emission from ABEI-GO@HRP was probably due to that HRP and GO facilitated the formation of O₂^•–, – CO₄^•2–, HO^•, and π-CC^• in the CL reaction, and GO as a reaction interface promoted the electron transfer of the radical-involved reaction. By virtue of ABEI-GO@HRP as a platform, an ultrasensitive, selective, and reagentless CL sensor was developed for H₂O₂ detection. The CL sensor exhibited a detection limit of 47 fM at physiological pH, which was more than 2 orders of magnitude lower than previously reported methods. This work reveals that bifunctionalization of GO by ABEI and HRP leads to excellent CL feature and enzyme selectivity, which can be used as an ideal platform for developing novel analytical methods

Acridinium Ester-Functionalized Carbon Nanomaterials: General Synthesis Strategy and Outstanding Chemiluminescence

In this work, three different kinds of acridinium ester (AE)-functionalized carbon nanomaterials, including AE-functionalized carbon nanoparticles (AE-CNPs), AE-functionalized graphene oxide (AE-GO), and AE-functionalized multiwalled carbon nanotubes (AE-MCNTs), were synthesized for the first time via a simple, general, and noncovalent strategy. AE molecules were assembled on the surface of carbon nanomaterials by electrostatic interaction, π–π stacking interaction, and amide bond. The synthesized AE-CNPs, AE-GO, and AE-MCNTs with 5.0 × 10^–8 mol·L^–1 of synthetic AE concentration, which was very low compared with other chemiluminescence (CL) reagents such as luminol, <i>N</i>-(aminobutyl)-<i>N</i>-(ethylisoluminol), and lucigenin at the concentration of 3.3 × 10^–4 to 5.0 × 10^–6 mol·L^–1 used for the synthesis of CL-functionalized nanomaterials, exhibited outstanding CL activity and good stability. It was found that carbon nanomaterials as nanosized platforms could efficiently immobilize AE molecules and facilitate the formation of OH^• and O₂^•–, leading to strong light emission. Moreover, the CL intensity of AE-GO was the highest, which was about 8.7 and 3.7 times higher than that of AE-CNPs and AE-MCNTs, respectively. This mainly resulted from a difference in the amount of adsorbed AE molecules on the surface of different carbon nanomaterials. Additionally, the prepared AE-CNPs demonstrated excitation-dependent fluorescence property and good fluorescence stability against photobleaching. On the basis of the excellent CL and special fluorescence properties of AE-CNPs, a dual-mode array strategy has been proposed for the first time and seven kinds of transition-metal ions could be successfully discriminated

A General Chemiluminescence Strategy for Measuring Aptamer–Target Binding and Target Concentration

Although much effort has been made for studies on aptamer–target interactions due to promising applications of aptamers in biomedical and analytical fields, measurement of the aptamer–target binding constant and binding site still remains challenging. Herein, we report a sensitive label-free chemiluminescence (CL) strategy to determine the target concentration and, more importantly, to measure the target–aptamer binding constant and binding site. This approach is suitable for multiple types of targets, including small molecules, peptides, and proteins that can enhance the CL initiated by <i>N</i>-(amino­butyl)-<i>N</i>-ethyliso­luminol functionalized gold colloids, making the present method a general platform to investigate aptamer–target interactions. This approach can achieve extremely high sensitivity with nanogram samples for measuring the target–aptamer binding constant. And the measurement could be rapidly performed using a simple and low-cost CL system. It provides an effective tool for studying the binding of biologically important molecules to nucleic acids and the selection of aptamers. Besides, we have also discovered that the 14-mer aptamer fragment itself split from the ATP-binding aptamer could selectively capture ATP. The binding constant, site, and conformation between ATP and the 14-mer aptamer fragment were obtained using such a novel CL strategy and molecular dynamic simulation

Micro-Nanostructured Polyaniline Assembled in Cellulose Matrix via Interfacial Polymerization for Applications in Nerve Regeneration

Conducting polymers have emerged as frontrunners to be alternatives for nerve regeneration, showing a possibility of the application of polyaniline (PANI) as the nerve guidance conduit. In the present work, the cellulose hydrogel was used as template to in situ synthesize PANI via the limited interfacial polymerization method, leading to one conductive side in the polymer. PANI sub-micrometer dendritic particles with mean diameter of ∼300 nm consisting of the PANI nanofibers and nanoparticles were uniformly assembled into the cellulose matrix. The hydrophobic PANI nanoparticles were immobilized in the hydrophilic cellulose via the phytic acid as “bridge” at presence of water through hydrogen bonding interaction. The PANI/cellulose composite hydrogels exhibited good mechanical properties and biocompatibility as well as excellent guiding capacity for the sciatic nerve regeneration of adult Sprague–Dawley rats without any extra treatment. On the basis of the fact that the pure cellulose hydrogel was an inert material for the neural repair, PANI played an indispensable role on the peripheral nerve regeneration. The hierarchical micro-nanostructure and electrical conductivity of PANI could remarkably induce the adhesion and guiding extension of neurons, showing its great potential in biomedical materials

Shackling Effect Induced Property Differences in Metallo-Supramolecular Polymers

We demonstrate here the synthesis of a novel class of metallo-supramolecular polymers with shackled structure, via the coordination of cyclic di­(bis-terpyridine-triphenyl ether ester) ligands with ruthenium­(II) ions. The constraint from the ring topology via the shackling of ligands provides novel properties to these metallo-supramolecular polymers, including the formation of dendritic crystals, red-shift of absorption bands in the UV–vis spectra from interchain charge-transfer transitions, and a typical flash-type memory behavior