Conjugation behaviours of CdTe quantum dots and antibody by a novel immunochromatographic method

Three water-soluble CdTe quantum dots (QDs) (green-emitting, yellow-emitting and red-emitting) were synthesised for different refluxing time with 3-mercaptopropionic acid (MPA) as stabiliser. Then the red-emitting CdTe QDs and mouse immunoglobulin G (IgG) were taken as the representative to study the conjugation behaviour of QDs and antibody by a novel immunochromatographic method. After comparing with several methods, that is, direct conjugation, 1-ethyl-3(3-dimethylaminopropyl) carbodiimides hydrochloride (EDC)-mediated conjugation, N-hydroxysuccinimide (NHS)-mediated conjugation, EDC/NHS-mediated conjugation by immunochromatographic strips, EDC and NHS were selected together as coupling agents to conjugate QDs with antibody efficiently. Finally, the K562 leukaemia cells were incubated with the EDC/NHS-mediated conjugates to evaluate the performance in practical application, and the result from fluorescence images showed that it was successfully applied to label cells. The immunochromatographic strip was a superior method to study the conjugation of the fluorophore and antibody.     

Characterization of the functional binding properties of antibody conjugated quantum dots

Antibody conjugated quantum dots are an emerging technology for high-resolution labeling of biological systems. In this work we determined the number of functional antibodies (i.e., antibodies that are sterically available for functional binding to target proteins) conjugated to semiconductor quantum dots. This is critical for the interpretation of biological data labeled with these methods. We found that the number of available functional antibodies varied significantly for different conjugation methods and are lower than previously estimated. These results may suggest potential strategies for improving quantum dot labeling of biological preparations.     

有缺陷和无缺陷量子点内应变分布的比较与分析

阐述了量子点内能级结构与应变的关系,用ANSYS7．1计算了有缺陷和无缺陷两种情况下：InAs／GaAs量子点的应变分布,通过对计算结果的比较,讨论了两种情况下不同的应变分布对量子点电子结构影响的不同结果,指出有缺陷时量子点各能级的改变量都与无缺陷时不同,无缺陷时应变的作用只是使能级平行移动;有缺陷时,缺陷将使量子点内能级结构复杂化,有缺陷时发光波长和发光光谱都比无缺陷时复杂。     

Conjugation of luminescent quantum dots with antibodies using an engineered adaptor protein to provide new reagents for fluoroimmunoassays

We describe the preparation and characterization of bioinorganic conjugates made with highly luminescent semiconductor CdSe-ZnS core-shell quantum dots (QDs) and antibodies for use in fluoroimmunoassays. The conjugation strategy employs an engineered molecular adaptor protein, attached to the QDs via electrostatic/hydrophobic self-assembly, to link the inorganic fluorophore with antibodies. In this method, the number of antibodies conjugated to a single QD can be varied. In addition, we have developed a simple purification strategy based on mixed-composition conjugates of the molecular adaptor and a second two-domain protein that allows the use of affinity chromatography. QD-antibody conjugates were successfully used in fluoroimmunoassays for detection of both a protein toxin (staphylococcal enterotoxin B) and a small molecule (2,4,6-trinitrotoluene).     

Single-domain antibody bioconjugated near-IR quantum dots for targeted cellular imaging of pancreatic cancer

Successful targeted imaging of BxPC3 human pancreatic cancer cells is feasible with near-IR CdTeSe/CdS quantum dots (QDs) functionalized with single-domain antibody (sdAb) 2A3. For specific targeting, sdAbs are superior to conventional antibodies, especially in terms of stability, aggregation, and production cost. The bright CdTeSe/CdS QDs were synthesized to emit in the diagnostic window of 650-900 nm with a narrow emission band. 2A3 was derived from llama and is small in size of 13 kDa, but with fully-functional recognition to the target carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6), a possible biomarker as a therapeutic target of pancreatic cancer. For compelling imaging, optical may be the most sensible among the various imaging modalities, regarding the sensitivity and cost. This first report on sdAb-conjugated near-IR QDs with high signal to background sensitivity for targeted cellular imaging brings insights into the development of optical molecular imaging for early stage cancer diagnosis.     

Luminescent quantum dots fluorescence resonance energy transfer-based probes for enzymatic activity and enzyme inhibitors

The paper describes the development and characterization of analytical properties of quantum dot-based probes for enzymatic activity and for screening enzyme inhibitors. The luminescent probes are based on fluorescence resonance energy transfer (FRET) between luminescent quantum dots that serve as donors and rhodamine acceptors that are immobilized to the surface of the quantum dots through peptide linkers. Peptide-coated CdSe/ZnS quantum dots were prepared using a one-step ligand exchange process in which RGDC peptide molecules replace trioctylphosphine oxide (TOPO) molecules as the capping ligands of the quantum dots. The peptide molecules were bound to the surface of the CdSe/ZnS quantum dots through the thiol group of the peptide cysteine residue. The peptide-coated quantum dots were labeled with rhodamine to form the FRET probes. The emission quantum yield of the quantum dot FRET probes was 4-fold lower than the emission quantum yield of TOPO-capped quantum dots. However, the quantum dot FRET probes were sufficiently bright to enable quantitative enzyme and enzyme inhibition assays. The probes were used first to test the enzymatic activity of trypsin in solution based on FRET signal changes of the quantum dot-based enzymatic probes in the presence of proteolytic enzymes. For example, exposure of the quantum dot FRET probes to 500 microg/mL trypsin for 15 min resulted in 60% increase in the photoluminescence of the quantum dots and a corresponding decrease in the emission of the rhodamine molecules. These changes resulted from the release of rhodamine molecules from the surface of the quantum dots due to enzymatic cleavage of the peptide molecules. The quantum dot FRET-based probes were used to monitor the enzymatic activity of trypsin and to screen trypsin inhibitors for their inhibition efficiency.     

Critical parameters for the scale-up synthesis of quantum dots

The optical properties of quantum dots strongly depend on the synthesis conditions including the reagent grade of the chemicals. We developed a synthesis set-up allowing the reproducible preparation of quantum dots with a high control over chemical and physical parameters. Here, we report on the effects of the injection speed, the stirring velocity and the reagent grade of the chemicals on the photoluminescence behaviour of CdSe quantum dots. Additionally we compared the commonly used hot-injection method with a novel one-pot approach leading to a slightly larger particle size distribution of the resulting high quality quantum dots in the latter case.     

Capillary electrophoresis method for the characterization and separation of CdSe quantum dots

This paper presents a simple and rapid methodology to separate and characterize free CdSe quantum dots (QDs) in aqueous medium by capillary electrophoresis (CE). First, we describe a controlled derivatization procedure to obtain water-soluble QDs through noncovalent interactions. This derivatization methodology was based on the formation of a complex between the QDs and several types of surfactants to enhance the hydrophilicity and stability of the CdSe QDs. The surfactants used to achieve the surface functionalization were trioctylphosphine oxide/trioctylphosphine (TOPO/TOP) and sodium dodecyl sulfate (SDS). Different CdSe QDs core sizes were synthesized as function of the nanocrystals growing time and then subjected to controlled coating. These free QDs were separated by capillary zone electrophoresis (CZE) based on the differences in the charge-to-mass ratio of the QDs-TOPO/TOP-SDS complexes, and the detection was carried out with UV-vis and laser-induced fluorescence (LIF) techniques obtaining detection limits 5 times lower with CE-LIF. Under the optimal working conditions, four different-sized QDs were successfully separated whose average sizes were 3.1, 3.6, 4.3, and 4.9 nm, and the size distribution was less than 7% for all of them [calculated from the full width at half-maximum (fwhm) of the fluorescence spectra and confirmed by high-resolution transmission electron microscopy (HTEM)]. Therefore, we were able to separate QDs that differ in only 0.5 nm in diameter and 19 nm in fluorescence emission maximum. This corresponds to the better resolution achieved in the analysis of these kinds of nanoparticles. Finally, a correlation between the migration times plus or minus peak width and the core sizes plus or minus size distribution was established.     

Preparation of EGFR monoclonal antibody conjugated nanoparticles and targeting to hepatocellular carcinoma

This study aims to determine the sensitivity, specificity and accuracy of epidermal growth factor receptor monoclonal antibody (EGFRmAb) modified poly(lactic acid-co-l-lysine) nanoparticles (PLA-PLL-EGFRmAb) NPs delivery system to EGFR positive cancer cells. In the study, a new PLA-PLL-EGFRmAb NPs was prepared. The cellular cytotoxicity, cellular uptake, and the targeted effect for hepatocellular carcinoma of PLA-PLL-EGFRmAb NPs were investigated. In vitro, the findings of Flow cytometry and Confocal Laser scanning Biological Microscopy showed that PLA-PLL-EGFRmAb NPs can bind to hepatocellular carcinoma cells and were uptaken effectively. In vivo in the SMMC-7721 xenograft mouse model, PLA-PLL-EGFRmAb NPs could target to the tumor effectively, which demonstrated a better targeting. These results showed that the PLA-PLL-EGFRmAb NPs have the potential to be used as a target delivery carrier for tumor therapies.     

Charge transport through a set of quantum dots with oscillating electron tunnelling rates between the quantum dots and the drain electron reservoir

For nanometer size electronic circuits utilizing mechanical elements such as beams or cantilevers with a single oscillating element, time-dependent electron tunnelling events can appear requiring a knowledge of the different characteristics of these systems. In this paper one such system has been studied. This is a set of six quantum dots (QDs) placed in parallel configuration between two electron reservoirs. The drain electrode is coupled with the quantum dots through time-dependent tunnelling matrix elements. Applying the equation of motion method for appropriate correlation functions, the current flowing in this system was calculated and its dependence on the parameters describing the quantum dots discussed.     

Synthesis and conductivity mapping of SnS quantum dots for photovoltaic applications

Quantum dots (QDs) are considered a possible solution to overcome the Shockley–Queisser efficiency limit of 31% for single junction solar cells by efficiently absorbing above band gap energy photons through Multiple Exciton Generation (MEG) or sub band gap energy photons using an Intermediate Band Solar Cell structure (IBSC). For the latter absorption process, we consider tin sulphide (SnS) as a promising candidate, having several advantages compared to the other nanoparticles studied extensively so far, such as CdS, CdSe, PbS, and PbSe; namely it is non-toxic and environmentally benign and thus will be most suitable in consumer products such as solar panels.In this work we propose a new colloidal synthesis method for SnS QDs. We have obtained mono-dispersive SnS and SnS/In₂S₃ core–shell nanoparticles with a size of ～4 nm. Energy dispersive X-ray spectroscopy (EDX) elemental analysis revealed that the particles are indeed SnS and not SnS₂. Furthermore, the conductive nature of the nanoparticles has been inferred by conductivity mapping using a relatively new contactless technique, Torsional Resonance Tunneling AFM (TR-TUNA). These results confirm that the SnS QDs possess all the requirements to be applied as photoactive layers in photovoltaic devices.     

Biofunctionalised quantum dots for sensing and identification of waterborne bacterial pathogens

Harmful bacteria are the most common cause of food- and waterborne illnesses. Infection often leads to bloody diarrhoea, and occasionally to kidney failure. Several strains of the bacteria Escherichia coli produce a powerful toxin which causes serious illness. Food and water can be contaminated with other bacteria like Salmonella, Coliform, Pseudomonas, etc. Hence, it has become important to rapidly detect and identify infectious bacteria. Colloidal luminescent semiconductor nanocrystals or quantum dots (QDs) have elicited a great deal of interest in the biosensing community due to their unique fluorescent properties. Here ZnS?:?Mn²⁺ QDs are synthesised and biofunctionalised with chitosan. They are attached to the anionic cell wall of E. coli bacteria and different properties of this compound system are studied. These nanocrystals may offer cost effective and quicker alternative to detect single bacterium compared to other conventional methods. The process of the synthesis of QDs, biofunctionalisation and detection of bacteria have been characterised by XRD, UV-Vis spectroscopy, FTIR, photoluminescence spectroscopy, AFM, high-resolution transmission electron microscopy and confocal microscopy. The particle size calculated is approximately 8–10?nm. The blue shift of PL peak has been observed after the bacteria get attached.     

CdSe and CdSe/ZnS quantum dots for the detection of C-reactive protein

In this study, a method for the detection of C-reactive protein (CRP) using CdSe and CdSe/ZnS quantum dots (QDs) is proposed. CdSe and CdSe/ZnS core-shell QDs are synthesised by using 2-mercaptosuccinic acid (MSA) as a capping agent. These QDs were then subjected to various characterisation studies, namely X-ray diffraction and transmission electron microscope for size and structure, Fourier transform infrared spectroscopy for the confirmation of functional groups, ultraviolet–visible absorption and fluorescence spectroscopy for optical characteristics and dynamic light scattering for hydrodynamic changes of QDs. Two biochemical mixtures were developed: one by mixing blood serum containing CRP and CdSe-phosphorylethanolamine (PEA) and the other by mixing blood serum with CdSe/ZnS-PEA. When these mixtures are observed for fluorescence due to interaction of QDs with CRP, a correlation between changes in fluorescence for different concentrations of CRP is noted. The result demonstrates that CRP can be detected with the help of QDs without using any antibodies.     

A novel method for the preparation of water-soluble and small-size CdSe quantum dots

A novel method for the preparation of water-soluble and small-size CdSe quantum dots has been reported under high-intensity ultrasonic irradiation. The as-prepared products have been characterized by absorption spectra, X-ray powder diffraction (XRD) and high-resolution transmission electron microscopy (HRTEM).     

Label-free fluorescent detection of protein kinase activity based on the aggregation behavior of unmodified quantum dots

Herein, we present a novel label-free fluorescent assay for monitoring the activity and inhibition of protein kinases based on the aggregation behavior of unmodified CdTe quantum dots (QDs). In this assay, cationic substrate peptides induce the selective aggregation of unmodified QDs with anionic surface charge, whereas phosphorylated peptides do not. Phosphorylation by kinase alters the net charge of peptides and subsequently inhibits the aggregation of unmodified QDs, causing an enhanced fluorescence with a 45 nm blue-shift in emission and a yellow-to-green emission color change. Hence the fluorescence response allows this QD-based method to easily probe kinase activity by a spectrometer or even by the naked eye. The feasibility of the method has been demonstrated by sensitive measurement of the activity of cAMP-dependent protein kinase (PKA) with a low detection limit (0.47 mU μL(-1)). On the basis of the fluorescence response of QDs on the concentration of PKA inhibitor H-89, the IC(50) value, the half maximal inhibitory concentration, was estimated, which was in agreement with the literature value. Moreover, the system can be applicable to detect the Forskolin/3-isobutyl-1-methylxantine (IBMX)-stimulated activation of PKA in cell lysate. Unlike the existing QD-based enzyme activity assays in which the modification process of QDs is essential, this method relies on unmodified QDs without the requirement of peptide labeling and QDs' modification, presenting a promising candidate for cost-effective kinase activity and inhibitor screening assays.     

CdSe-ZnS quantum dots for selective and sensitive detection and quantification of hypochlorite

Four types of water-soluble quantum dots (QDs) grafted with different organic coating layers were fabricated, and their sensitivities for hypochlorite/hypochlorous acid (HClO) were examined. It was found that QDs with HClO reactive (methylamino and sulphide groups) coating layers exhibited a protective effect on HClO quenching of QD fluorescence, whereas QDs with hydrocarbon and carboxylate coating layers showed least protection to QD fluorescence quenching by HClO and, thus, has the highest sensitivity for the detection of HClO. The QDs with carboxylate coating layers (QDs-poly-CO(2)(-)) was successfully applied to the quantification of HClO in tap water. The excellent selectivity of the QDs-poly-CO(2)(-) toward hypochlorite against other reactive oxygen species allowed us to monitor myeloperoxidase activity. Finally, the QDs-poly-CO(2)(-) was also used for the detection of hypochlorite in HL60 cells by fluorescent imaging. Hence, QD-poly-CO(2)(-) exhibits great promise as a selective and sensitive HClO probe in chemical and biological systems.     

Bio-templated CdSe quantum dots green synthesis in the functional protein,lysozyme, and biological activity investigation

Bifunctional fluorescence (CdSe Quantum Dots) – protein (Lysozyme) nanocomposites were synthesized at room temperature by a protein-directed, solution-phase, green-synthetic method. Fluorescence (FL) and absorption spectra showed that CdSe QDs were prepared successfully with Lyz. The average particle size and crystalline structure of QDs were investigated by high-resolution transmission electron microscopy (HRTEM) and X-ray diffraction (XRD), respectively. With attenuated total reflection-fourier transform infrared (ATR-FTIR) spectra and thermogravimetric (TG) analysis, it was confirmed that there is interaction between QDs and amide I, amide II groups in Lyz. FL polarization was measured and FL imaging was done to monitor whether QDs could be responsible for possible changes in the conformation and activity of Lyz. Interestingly, the results showed Lyz still retain the biological activity after formation of QDs, but the secondary structure of the Lyz was changed. And the advantage of this synthesis method is producing excellent fluorescent QDs with specifically biological function.     

Method for determining the elemental composition and distribution in semiconductor core-shell quantum dots

In the biological sciences, the use of core-shell quantum dots (QDs) has gained wide usage but analytical challenges still exist for characterizing the QD structure. The application of energy-dispersive X-ray spectroscopy and X-ray photoelectron spectroscopy (XPS) to bulk materials is relatively straightforward; however, for meaningful applications of surface science techniques to multilayer nanoparticles requires novel modifications and analysis methods. To experimentally characterize the elemental composition and distribution in CdSe/CdS/ZnS QDs, we first develop a XPS signal subtraction technique capable of separating the overlapped selenium 3s (core) and sulfur 2s (shell) peaks (both peaks have binding energies near 230 eV) with higher precision than is typically reported in the nanoparticle literature. This method is valid for any nanoparticle containing selenium and sulfur. Then we apply a correction formula to the XPS data and determine that the 2 nm stoichiometric CdSe core is surrounded by 2 CdS layers and a stoichimetric ZnS monolayer. These findings and the multiapproach methodology represent a significant advancement in the detailed surface science study of multilayer nanoparticles. In agreement with recent surprising findings, the time-of-flight secondary mass spectrometry measurements suggest that the surface sites of the QDs used in this study are primarily covered with a mixture of octadecylphosphonic acid and trioctylphophine oxide.     

Preparation of AgInS2 quantum dots and their application for trypsin detection

Journal of Materials Science: Materials in Electronics - Water-soluble AgInS2 quantum dots were prepared by hot injection method using glutathione (GSH) as the stabilizer. The obtained AIS QDs were...     

Surface functionalized cadmium telluride quantum dots for the optical detection and determination of herbicides

An optical sensor for detection of herbicides was developed through the functionalization of CdTe quantum dots (CdTe-QDs) with cysteamine hydrochloride. The functionalized CdTe-QDs was characterized with various physicochemical methods such as X-ray diffraction, transmission electron microscopy, Fourier transform infrared spectroscopy, energy dispersive X-ray analysis, Ultraviolet–visible and photoluminescence spectroscopies. The optical band gap of the functionalized CdTe-QDs as calculated by using Tauc plot was 3.75 eV. It was found that the fluorescence intensity of the functionalized CdTe-QDs quenched linearly in the presence of different herbicides according to the Stern–Volmer equation. Thus, the functionalized CdTe-QDs can be used as simple, rapid, inexpensive, and optical sensitive sensor for practical detection of herbicides.