Overview of the Design and Application of Photothermal Immunoassays.

Developing powerful immunoassays for sensitive and real-time detection of targets has always been a challenging task. Due to their advantages of direct readout, controllable sensing, and low background interference, photothermal immunoassays have become a type of new technology that can be used for various applications such as disease diagnosis, environmental monitoring, and food safety. By modification with antibodies, photothermal materials can induce temperature changes by converting light energy into heat, thereby reporting specific target recognition events. This article reviews the design and application of photothermal immunoassays based on different photothermal materials, including noble metal nanomaterials, carbon-based nanomaterials, two-dimensional nanomaterials, metal oxide and sulfide nanomaterials, Prussian blue nanoparticles, small organic molecules, polymers, etc. It pays special attention to the role of photothermal materials and the working principle of various immunoassays. Additionally, the challenges and prospects for future development of photothermal immunoassays are briefly discussed.

Gold Nanoparticles-Based Colorimetric Immunoassay of Carcinoembryonic Antigen with Metal-Organic Framework to Load Quinones for Catalytic Oxidation of Cysteine.

This work reported gold nanoparticles (AuNPs)-based colorimetric immunoassay with the Cu-based metal-organic framework (MOF) to load pyrroloquinoline quinone (PQQ) for the catalytic oxidation of cysteine. In this method, both Cu2+ and PQQ in the MOF could promote the oxidation of inducer cysteine by redox cycling, thus limiting the cysteine-induced aggregation of AuNPs and achieving dual signal amplification. Specifically, the recombinant carcinoembryonic antigen (CEA) targets were anchored on the MOF through the metal coordination interactions between the hexahistidine (His6) tag in CEA and the unsaturated Cu2+ sites in MOF. The CEA/PQQ-loaded MOF could be captured by the antibody-coated ELISA plate to catalyze the oxidation of cysteine. However, once the target CEA in the samples bound to the antibody immobilized on the plate surface, the attachment of CEA/PQQ-loaded MOF would be limited. Cysteine remaining in the solution would trigger the aggregation of AuNPs and cause a color change from red to blue. The target concentration was positively related to the aggregation and color change of AuNPs. The signal-on competitive plasmonic immunoassay exhibited a low detection limit with a linear range of 0.01-1 ng/mL. Note that most of the proteins in commercial ELISA kits are recombinant with a His6 tag in the N- or C-terminal, so the work could provide a sensitive plasmonic platform for the detection of biomarkers.

Half-decomposition of salt-bearing dolomite.

Half-calcined dolomites (HCDs) have been widely used in environmental remediation, medicine, and construction. However, advanced calcination technologies are required to modify their microstructure and thus improve their working performance. Herein, we investigated the effects of a variety of inorganic salts on the decomposition of dolomite based on thermogravimetric, compositional, and morphological analysis. The thermogravimetric data showed that certain salts significantly lowered the half-decomposition temperature of dolomite, which included LiCl, CaCl2, MgCl2, AlCl3, LiNO3, KNO3, K2CO3, Li2CO3, Li2SO4, Na3PO4, and K3PO4. Compositional analysis demonstrated that only half-decomposition occurred when salt-bearing dolomite was calcined at a temperature of 723-923 K, leading to the formation of CaO-free HCDs composed of periclase and Mg-calcite having a Mg level of 2.0-10.5 mol%. Morphological analysis showed that porous HCDs were feasibly obtained by calcining salt-bearing dolomite at 723-923 K. MgO coarsening occurred at a temperature above 873 K, but it could be avoided by controlling the calcination time. The mechanism of salts may be related to the heterovalent doping effect, which may lead to an increase in the concentration of vacancies in the dolomite lattice.

Advancements in Biosensors Based on the Assembles of Small Organic Molecules and Peptides.

Over the past few decades, molecular self-assembly has witnessed tremendous progress in a variety of biosensing and biomedical applications. In particular, self-assembled nanostructures of small organic molecules and peptides with intriguing characteristics (e.g., structure tailoring, facile processability, and excellent biocompatibility) have shown outstanding potential in the development of various biosensors. In this review, we introduced the unique properties of self-assembled nanostructures with small organic molecules and peptides for biosensing applications. We first discussed the applications of such nanostructures in electrochemical biosensors as electrode supports for enzymes and cells and as signal labels with a large number of electroactive units for signal amplification. Secondly, the utilization of fluorescent nanomaterials by self-assembled dyes or peptides was introduced. Thereinto, typical examples based on target-responsive aggregation-induced emission and decomposition-induced fluorescent enhancement were discussed. Finally, the applications of self-assembled nanomaterials in the colorimetric assays were summarized. We also briefly addressed the challenges and future prospects of biosensors based on self-assembled nanostructures.

Simple, sensitive and selective detection of dopamine using dithiobis(succinimidylpropionate)-modified gold nanoparticles as colorimetric probes.

In this paper, we report a simple, sensitive and selective colorimetric visualization of dopamine (DA) using dithiobis(succinimidylpropionate) (DSP)-modified gold nanoparticles (AuNPs) as probes and ferric ions as cross-linkers. Via the standard amine coupling reaction between the amino groups of DA and activated carboxyl groups of DSP, DA molecules can be assembled onto the surface of DSP-AuNPs. Accordingly, Fe(3+) ions induce a change of DSP-AuNPs in color and UV-vis absorbance by coordinating to the catechol groups of the anchored DA. The pH dependence and mechanism of this method are discussed. A detection limit of 2 nM was obtained, which is lower than those achievable with currently used chromatographic and electrochemical techniques. The feasibility for the detection of DA in artificial cerebrospinal fluid has been demonstrated.

Simple, fast and selective detection of adenosine triphosphate at physiological pH using unmodified gold nanoparticles as colorimetric probes and metal ions as cross-linkers.

We report a simple, fast and selective colorimetric assay of adenosine triphosphate (ATP) using unmodified gold nanoparticles (AuNPs) as probes and metal ions as cross-linkers. ATP can be assembled onto the surface of AuNPs through interaction between the electron-rich nitrogen atoms and the electron-deficient surface of AuNPs. Accordingly, Cu2+ ions induce a change in the color and UV/Vis absorbance of AuNPs by coordinating to the triphosphate groups and a ring nitrogen of ATP. A detection limit of 50 nM was achieved, which is comparable to or lower than that achievable by the currently used electrochemical, spectroscopic or chromatographic methods. The theoretical simplicity and high selectivity reported herein demonstrated that AuNPs-based colorimetric assay could be applied in a wide variety of fields by rationally designing the surface chemistry of AuNPs. In addition, our results indicate that ATP-modified AuNPs are less stable in Cu2+, Cd2+ or Zn2+-containing solutions due to the formation of the corresponding dimeric metal-ATP complexes.

Overview on the Design of Magnetically Assisted Electrochemical Biosensors.

Electrochemical biosensors generally require the immobilization of recognition elements or capture probes on the electrode surface. This may limit their practical applications due to the complex operation procedure and low repeatability and stability. Magnetically assisted biosensors show remarkable advantages in separation and pre-concentration of targets from complex biological samples. More importantly, magnetically assisted sensing systems show high throughput since the magnetic materials can be produced and preserved on a large scale. In this work, we summarized the design of electrochemical biosensors involving magnetic materials as the platforms for recognition reaction and target conversion. The recognition reactions usually include antigen-antibody, DNA hybridization, and aptamer-target interactions. By conjugating an electroactive probe to biomolecules attached to magnetic materials, the complexes can be accumulated near to an electrode surface with the aid of external magnet field, producing an easily measurable redox current. The redox current can be further enhanced by enzymes, nanomaterials, DNA assemblies, and thermal-cycle or isothermal amplification. In magnetically assisted assays, the magnetic substrates are removed by a magnet after the target conversion, and the signal can be monitored through stimuli-response release of signal reporters, enzymatic production of electroactive species, or target-induced generation of messenger DNA.

Automatic discrimination of the geographical origins of milks by excitation-emission fluorescence spectrometry and chemometrics.

This paper presents the automatic discrimination of geographical origins of milks from Western Yunnan Plateau areas and eastern China by excitation-emission fluorescence spectrometry and chemometrics. Genuine plateau milks (n = 60) and milks from eastern China (n = 89) are scanned in the regions of 180-300 nm for excitation and 200-800 nm for emission. Different options of data analysis are investigated and compared in terms of their performance in discriminating milks of different geographical origins: (1) two-way partial least squares discriminant analysis (PLSDA) based on excitation and emission spectra, respectively; (2) two-way PLSDA based on fusion of excitation and emission spectra; (3) three-way PLSDA based on excitation-emission matrix spectra. The two-way PLSDA methods with excitation spectra, emission spectra, and fusion of excitation and emission spectra correctly classify 91.3%, 88.6%, and 95.3% of the milk samples, respectively; while the total accuracy of three-way PLSDA is 96.0%. The results demonstrate the two-way data combining excitation and emission spectra are sufficient to characterize and identify the plateau milks. Considering both model accuracy and the analytical time required, two-way PLS-DA with fusion of excitation and emission spectra is recommended as a reliable and quick method to discriminate plateau milks from ordinary milks.

Electrochemical Immunosensors with PQQ-Decorated Carbon Nanotubes as Signal Labels for Electrocatalytic Oxidation of Tris(2-carboxyethyl)phosphine.

Nanocatalysts are a promising alternative to natural enzymes as the signal labels of electrochemical biosensors. However, the surface modification of nanocatalysts and sensor electrodes with recognition elements and blockers may form a barrier to direct electron transfer, thus limiting the application of nanocatalysts in electrochemical immunoassays. Electron mediators can accelerate the electron transfer between nanocatalysts and electrodes. Nevertheless, it is hard to simultaneously achieve fast electron exchange between nanocatalysts and redox mediators as well as substrates. This work presents a scheme for the design of electrochemical immunosensors with nanocatalysts as signal labels, in which pyrroloquinoline quinone (PQQ) is the redox-active center of the nanocatalyst. PQQ was decorated on the surface of carbon nanotubes to catalyze the electrochemical oxidation of tris(2-carboxyethyl)phosphine (TCEP) with ferrocenylmethanol (FcM) as the electron mediator. With prostate-specific antigen (PSA) as the model analyte, the detection limit of the sandwich-type immunosensor was found to be 5 pg/mL. The keys to success for this scheme are the slow chemical reaction between TCEP and ferricinum ions, and the high turnover frequency between ferricinum ions, PQQ. and TCEP. This work should be valuable for designing of novel nanolabels and nanocatalytic schemes for electrochemical biosensors.

Duplex-specific nuclease-based electrochemical biosensor for the detection of microRNAs by conversion of homogeneous assay into surface-tethered electrochemical analysis.

We proposed a simple and sensitive strategy for the detection of microRNAs (miRNAs) by converting homogeneous assay into surface-tethered electrochemical analysis. Specifically, the biotinylated detection probes (biotin-DNA-biotin) can trigger the in-situ assembly of tetrameric streptavidin (SA) proteins on an electrode surface via the SA-biotin interactions. The (SA-biotin-DNA-biotin)n assemblies electrically insulated the electrode interface, thereby blocking the electron transfer of [Fe(CN)6]3-/4-. When the probe was hybridized with the target miRNA, it would be cleaved into small fragments (denoted as biotin-DNA) by duplex-specific nuclease (DSN). The released target miRNA can enter into the next hybridization-enzymolysis cycle, thus leading to the generation of considerable amounts of biotin-DNA fragments. The released biotin-DNA competed with the detection probe to bind SA, thus limiting the in-situ formation of (SA-biotin-DNA-biotin)n assemblies. The surface-tethered electrochemical analysis by the dual signal amplification of DSN and (SA-biotin-DNA-biotin)n assemblies has been used for the determination of miRNAs in cell lysate with a satisfactory result. The method showed a detection limit down to 10 aM. The "one-step" immobilization-free strategy can be used to design novel biosensors for the detection of other biomarkers.

Nanomaterials-Based Colorimetric Immunoassays.

Colorimetric immunoassays for tumor marker detection have attracted considerable attention due to their simplicity and high efficiency. With the achievements of nanotechnology and nanoscience, nanomaterials-based colorimetric immunoassays have been demonstrated to be promising alternatives to conventional colorimetric enzyme-linked immunoassays. This review is focused on the progress in colorimetric immunoassays with the signal amplification of nanomaterials, including nanomaterials-based artificial enzymes to catalyze the chromogenic reactions, analyte-induced aggregation or size/morphology change of nanomaterials, nanomaterials as the carriers for loading enzyme labels, and chromogenic reactions induced by the constituent elements released from nanomaterials.

Gold nanoparticle-based colorimetric method for the detection of prostate-specific antigen.

BACKGROUND: Prostate-specific antigen (PSA), a serine protease, is a biomarker for preoperative diagnosis and screening of prostate cancer and monitoring of its posttreatment. METHODS: In this work, we reported a colorimetric method for clinical detection of PSA using gold nanoparticles (AuNPs) as the reporters. The method is based on ascorbic acid (AA)-induced in situ formation of AuNPs and Cu2+-catalyzed oxidation of AA. Specifically, HAuCl4 can be reduced into AuNPs by AA; Cu2+ ion can catalyze the oxidation of AA by O2 to inhibit the formation of AuNPs. In the presence of the PSA-specific peptide (DAHSSKLQLAPP)-modified gold-coated magnetic microbeads (MMBs; denoted as DAHSSKLQLAPP-MMBs), complexation of Cu2+ by the MMBs through the DAH-Cu2+ interaction depressed the catalyzed oxidation of AA and thus allowed for the formation of red AuNPs. However, once the peptide immobilized on the MMB surface was cleaved by PSA, the DAHSSKLQ segment would be released. The resultant LAPP fragment remaining on the MMB surface could not sequestrate Cu2+ to depress its catalytic activity toward AA oxidation. Consequently, no or less AuNPs were generated. RESULTS: The linear range for PSA detection was found to be 0~0.8 ng/mL with a detection limit of 0.02 ng/mL. Because of the separation of cleavage step and measurement step, the interference of matrix components in biological samples was avoided. CONCLUSION: The high extinction coefficient of AuNPs facilitates the colorimetric analysis of PSA in serum samples. This work is helpful for designing of other protease biosensors by matching specific peptide substrates.

A Colorimetric Enzyme-Linked Immunosorbent Assay with CuO Nanoparticles as Signal Labels Based on the Growth of Gold Nanoparticles In Situ.

A colorimetric immunoassay has been reported for prostate-specific antigen (PSA) detection with CuO nanoparticles (CuO NPs) as signal labels. The method is based on Cu2+-catalyzed oxidation of ascorbic acid (AA) by O2 to depress the formation of colored gold nanoparticles (AuNPs). Specifically, HAuCl4 can be reduced by AA to produce AuNPs in situ. In the presence of target, CuO NPs-labeled antibodies were captured via the sandwich-type immunoreaction. After dissolving CuO nanoparticles with acid, the released Cu2+ catalyzed the oxidation of AA by O2, thus depressing the generation of AuNPs. To demonstrate the accuracy of the colorimetric assay, the released Cu2+ was further determined by a fluorescence probe. The colorimetric immunoassay shows a linear relationship for PSA detection in the range of 0.1~10 ng/mL. The detection limit of 0.05 ng/mL is comparable to that obtained by other CuO NPs-based methods. The high throughput, simplicity, and sensitivity of the proposed colorimetric immunoassay exhibited good applicability for assays of serum samples.

Sandwich-type electrochemical biosensor for glycoproteins detection based on dual-amplification of boronic acid-gold nanoparticles and dopamine-gold nanoparticles.

Glycoproteins play important roles in a wide variety of biological processes. The change in the concentration levels has been associated with many cancers, as well as other diseases. Thus, rapid, sensitive and selective determination of glycoproteins is much preferred. In this work, we reported a sandwich-type electrochemical biosensor based on dual-amplification of 4-mercaptophenylboronic acid (MBA)-capped gold nanoparticles (MBA-AuNPs) and dopamine (DA)-capped AuNPs (DA-AuNPs). Biological recognition elements such as synthetic receptor and aptamer immobilized onto gold electrodes were used to capture glycoproteins. The captured glycoproteins were then derivatized with MBA-AuNPs through the formation of tight covalent bonds between the boronic acids of MBA-AuNPs and diols of glycoproteins. Electroactive DA-AuNPs were attached by the anchored MBA-AuNPs via the interaction of boronic acids with DA tags, which facilities the amplified voltammetric detection of glycoproteins. With avidin and prostate specific antigen (PSA) as model analytes, we demonstrated the feasibility and sensitivity of the proposed method. The results indicated that sub-picomolar avidin/PSA can be readily measured. We believe that this strategy will be valuable for the electrochemical detection of other glycoproteins.

Predicting the age and type of tuocha tea by fourier transform infrared spectroscopy and chemometric data analysis.

Fourier transform infrared (FTIR) spectroscopy combined with chemometric multivariate methods was proposed to discriminate the type (unfermented and fermented) and predict the age of tuocha tea. Transmittance FTIR spectra ranging from 400 to 4000 cm(-1) of 80 fermented and 98 unfermented tea samples from Yunnan province of China were measured. Sample preparation involved finely grinding tea samples and formation of thin KBr disks (under 120 kg/cm(2) for 5 min). For data analysis, partial least-squares (PLS) discriminant analysis (PLSDA) was applied to discriminate unfermented and fermented teas. The sensitivity and specificity of PLSDA with first-derivative spectra were 93 and 96%, respectively. Multivariate calibration models were developed to predict the age of fermented and unfermented teas. Different options of data preprocessing and calibration models were investigated. Whereas linear PLS based on standard normal variate (SNV) spectra was adequate for modeling the age of unfermented tea samples (RMSEP = 1.47 months), a nonlinear back-propagation-artificial neutral network was required for calibrating the age of fermented tea (RMSEP = 1.67 months with second-derivative spectra). For type discrimination and calibration of tea age, SNV and derivative preprocessing played an important role in reducing the spectral variations caused by scattering effects and baseline shifts.