Fluorescence Resonance Energy Transfer System Research Articles

Evaluation of the Interaction between Bax and Hsp70 in Cells by Using a FRET System Consisting of a Fluorescent Amino Acid and YFP as a FRET Pair

To gain insight into factors that lead to dissociation of Bax from a complex with Hsp70 during apoptosis, we recently constructed a fluorescence resonance energy transfer (FRET) system composed of the Hsp70-YFP (YFP=yellow fluorescent protein) fusion protein and fluorescent amino acid (ANAP=6-acetyl(naphthalen-2-ylamino)-2-aminopropanoic acid)-containing Bax (Bax-ANAP), which was produced by using the genetic code expansion technique. In the current study, the FRET system was employed to elucidate how brefeldin A (an endoplasmic reticulum stress inducer), chlorpromazine and apoptozole (lysosomal membrane destabilizers), bafilomycin A1 (an inhibitor of lysosomal acidification) as well as raptinal and Az-TPP-O3 (mitochondria-targeted apoptosis inducers) affect the interaction between Bax and Hsp70. Analyses of single live cell images together with results of co-immunoprecipitation assays reveal that brefeldin A, chlorpromazine, and apoptozole promote dissociation of the Bax/Hsp70 complex through activation of the activator BH3-only protein. However, the results show that bafilomycin A1, raptinal, and Az-TPP-O3 have no influence on the interaction of Bax with Hsp70. The combined observations made in the current and previous studies demonstrate that the FRET system consisting of Bax-ANAP and Hsp70-YFP is highly useful to understand apoptotic processes associated with the Bax-Hsp70 interaction.

Smart “on-off-on” fluorescent switches for drug visual loading and responsive delivery

The unsatisfactory therapeutic effects and limitations of chemotherapy are urging people to find more effective therapy methods, and drug delivery systems have been attracting increasing research interests. However, most of these studies mainly focused on the therapeutic effects, while little attention was paid to explore new drug loading patterns and improve the efficiency. Herein, a pair of smart “on-off-on” fluorescent switches based on a novel double fluorescence resonance energy transfer (FRET) system was established to realize facilely visual drug loading and responsive delivery. To build this nanosystem, highly luminescent carbon quantum dots (CQDs) were first prepared and further loaded by anticancer drugs of doxorubicin (DOX). The π-π and hydrogen-bonding interactions between them readily formed the first FRET system, which allowed to facilely monitor the drug loading process by detecting the solution color changes with naked eyes. Then, manganese dioxide (MnO2) with high surface area, further modified by polydopamine and folic acid was synthesized and served as the other FRET receptor for CQDs and DOX, facilitating monitor of full drug absorption and loading by total fluorescence quenching. Moreover, when the double-FRET nanosystem was endocytosed into cancer cells, MnO2 was responsively degraded into Mn2+ under intracellular glutathione and acid conditions, which resulted into sustained drug release and gradual fluorescence recovery. More importantly, the constructed nano-system also exhibited selectively drug delivery and showed much finer therapeutic effects than single chemotherapy, holding great potential in future cancer therapy.

Simultaneous quantitative detection of multiple tumor markers in microfluidic nanoliter-volume droplets

We developed a simultaneous detection method for multiple tumor markers (TMs) in microfluidic droplets based on a multiple fluorescence resonance energy transfer (FRET) system. In this system, graphene oxide (GO) was used as the single quencher, while the multi-color quantum dots (QDs) labeled on different aptamers were employed as energy donors. When the aptamers were adsorbed onto GO due to the π-π stacking interaction, QDs were drawn to the surface of GO and quenched by it. Once the TMs were introduced, the corresponding fluorescence of QDs was recovered obviously owing to the preferential interaction of aptamers with the TMs. Here, the multi-FRET system was encapsulated into nanoliter-volume droplets by a simple T-junction microfluidic chip. The targets could be detected rapidly as the generated droplets flew through the integrated on-line detection zone. Three tumor markers, carcinoembryonic antigen (CEA), prostate-specific antigen (PSA) and vascular endothelial growth factor (VEGF165) could be detected simultaneously in 33 nL-volume droplets, which is only 1/3000 of the volume of the sample consumed in the conventional fluorescence spectrophotometer. In addition, the signals corresponding to different TM targets in one nanoliter-volume droplet could be read out at the same time, and the signals could be output continuously owing to the uninterruptible generation of droplets. Even with a signal acquisition frequency of 55 droplets per minute, the multi-FRET biosensing system has linear ranges of 0.50–70 ng mL−1 for CEA, 0.25–70 ng mL−1 for PSA and 0.50–70 ng mL−1 for VEGF165. The detection limits of CEA, PSA and VEGF165 were calculated to be 0.15 ng mL−1, 0.035 ng mL−1 and 0.11 ng mL−1, respectively. The method was also validated by analyzing human serum sample dilutions. The proposed multi-FRET-based system has potential to become a powerful tool for rapid, low-cost and simultaneous detection of multiple tumor markers.

A Versatile Photoinduced Electron Transfer‐Based Upconversion Fluorescent Biosensing Platform for the Detection of Disease Biomarkers and Nerve Agent

AbstractUpconversion nanoparticles (UCNPs) have emerged to be a new family of fluorescent probes for bioanalytical applications. In a typical design, the UCNPs act as the energy donors in a fluorescence resonance energy transfer (FRET) system, in which the target molecules mediate the energy transfer from the UCNPs to the acceptors, and their quantity information is consequently converted into the “on‐off” upconverting signals for readout. However, each UCNP contains thousands of emitting center ions and most of them are beyond the FRET critical distance, which hinders the fluorescence energy transfer efficiency, resulting in a low signal‐to‐background ratio (SBR). Herein, a new design is presented in which the energy of UCNPs is transferred to the o‐quinones on their surface via the photoinduced electron transfer (PET) mechanism. In this system, the quenching efficiency of UCNPs' fluorescence can be up to 94.73%, providing a high SBR. The performance of the PET‐based design is systematically testified, and the high‐sensitivity detection of disease biomarkers (tyrosinase and alkaline phosphatase) is demonstrated. Moreover, this UCNP‐PET platform is also capable of sensing the simulant of nerve agent sarin. This work will pave new ways to the design of UCNP‐based platforms toward bioanalytical applications.

Highly Stable Core-Shell Structured Semiconducting Polymer Nanoparticles for FRET-Based Intracellular pH Imaging.

Highly stable semiconducting polymer nanoparticles (NPs) (poly(9,9-dioctylfluorenyl-2,7-diyl) (PFO)/ poly(fluorene-2,7-ylenevinylene-co-phenylene) (PFV)-dopamine (DA) NPs) with previously unreported core-shell structure are developed for ratiometric sensing of intracellular pH values. PFO/PFV-DA NPs comprise central polyfluorene (PFO) as donor and PFV as acceptor, in which the donor and acceptor are spatially separated into the central core and nanoparticle shell. Specifically, thick PFV shells can not only significantly minimize the quenching interference of dopamine on the emission of standard reference (PFO), but are also able to maximize its accessibility to pH-sensitive dopamine and lead to sensitive response to pH changes. The resulting core-shell PFO/PFV NPs are structurally and optically stable, which can avoid the photobleaching and leakage of materials issues compared to traditional semiconducting polymer nanoparticles (SPNs)-based fluorescence resonance energy transfer (FRET) systems containing small molecules. Additionally, the designed compact PFO/PFV-DA NPs show quantitative response to the pH values in aqueous media and are capable of mapping intracellular pH fluctuations by ratiometric imaging. This work may open up opportunities for the generalizability of the consistent ratiometric emission intensity strategy based on core-shell structured SPNs nanoprobes for highly sensitive biological sensing.

A fluorometric aptamer nanoprobe for alpha-fetoprotein by exploiting the FRET between 5-carboxyfluorescein and palladium nanoparticles.

Alpha-fetoprotein (AFP) is a reliable clinical marker of hepatocellular carcinoma (HCC). A highly sensitive fluorometric aptamer nanoprobe is described for AFP detection. It is based on fluorescence resonance energy transfer (FRET) between AFP aptamer labelled with 5-carboxyfluorescein (FAM) and palladium nanoparticles (PdNPs). The PdNPs quench the green fluorescence of the FAM-AFP aptamer via interactions between nitrogen functional groups of the AFP aptamer and PdNPs. When AFP was introduced into the FAM-AFP aptamer-PdNPs FRET system, the AFP aptamer preferentially combines with AFP. This results in a conformational change and weakens the interaction between the aptamer and the PdNPs. Thus, the fluorescence of FAM recovers. The fluorescence recovery of FAM increases linearly in the 5.0-150ng·mL-1 AFP concentration range and has a 1.4 ng·mL-1 detection limit. The assay was applied to the analysis of spiked diluted human serum. The recovery values ranged from 98.3 to 112.9%, with relative standard deviations of <1.1%. This biosensing strategy provides a reliable and ultrasensitive protocol for the quantification of biomarkers with relevant antigens and aptamers. Graphical abstract Schematic presentation of a fluorometric aptamer nanoprobe for AFP assay based on fluorescence resonance energy transfer (FRET) between AFP aptamer labelled with 5-carboxyfluorescein (FAM) and palladium nanoparticles (PdNPs).

Intracellular Zinc Quantification by Fluorescence Imaging with a FRET System.

Fluorescence imaging of cellular metals is widely reported. However, the quantification of intracellular metals with fluorescence imaging is so far not feasible and highly challenging. In this work, a ratiometric probe with two fluorescently labeled complementary DNA strains is designed for intracellular zinc quantification via fluorescence imaging, based on fluorescence resonance energy transfer (FRET) from carbon dots (CDs) to fluorescein (FAM). The donor CDs are modified with a Zn2+ aptamer, whereas the receptor FAM is conjugated with the complementary DNA sequence to ensure selectivity. MCF-7 cells are cultured sequentially with Zn2+ (20, 40, 55, 70, 85, and 100 μmol L-1) and CDs-FAM (100 μg mL-1), which is used for fluorescence imaging (at λex = 405 nm and λem = 440-490 nm for CDs, λem = 500-550 nm for FAM) to provide a relative fluorescence ratio (( F - F0)/ F0, F = ICDs/ IFAM), followed by quantifying intracellular zinc with ICPMS. A linear correlation is achieved between the relative fluorescence ratio in fluorescence images and the intracellular zinc content derived by ICPMS, which facilitates intracellular zinc quantification via fluorescence imaging. It is especially useful for real-time tracing of intracellular zinc during the cell culturing process or in vivo. The cellular uptake of Zn2+ by MCF-7 cells is further evaluated with this approach by culturing with 100 μmol L-1 of Zn2+ for different times, and a maximum uptake of 60.5 fg per cell is observed at an incubation time of 60 min. This value is further demonstrated well by ICPMS detection.

Analysis of Protein-Protein Interaction in a Single Live Cell by Using a FRET System Based on Genetic Code Expansion Technology.

Hsp70 is known to directly bind to Bax for suppression of apoptosis. However, mechanisms on how Bax is dissociated from its complex with Hsp70 during apoptosis remain largely unknown. In the current study, we developed the efficient fluorescence resonance energy transfer (FRET) system which consisted of Hsp70-YFP and fluorescent amino acid (ANAP)-incorporated Bax, which was generated by using genetic code expansion technology, and applied the FRET system to elucidate mechanisms on how apoptosis-inducing substances dissociate Bax from Hsp70. Time-dependent analysis of single live cell images showed that Bax activators binding to Bax trigger sites inhibited the Bax-Hsp70 interaction but a Bax activator, which blocks phosphorylation of S184 via binding to the C-terminal S184 site, did not affect this interaction. Additionally, an inhibitor for Hsp70-Hsp40 interaction blocked the Bax-Hsp70 interaction. Furthermore, p53 activators promoted the dissociation of Bax from Hsp70 by reactivating p53 which disrupted the Bax-Hsp70 interaction. We also found that death ligands and a Bcl-2 inhibitor enhanced the dissociation of Bax from Hsp70 by activating activator BH3-only proteins. Results from this effort suggest that FRET systems consisting of the ANAP-incorporated protein and the YFP fusion protein will be valuable tools to gain an understanding of other types of protein-protein interactions.

A simple FRET system using two‐color CdTe quantum dots assisted by cetyltrimethylammonium bromide and its application to Hg(II) detection

In this study, we developed a novel simple fluorescence resonance-energy transfer (FRET) system between two-color CdTe quantum dots (QDs) assisted by cetyltrimethylammonium bromide (CTAB). Mercaptopropionic (MPA)-capped CdTe QDs serving as both donors and acceptors were successfully synthesized by changing the refluxing time in aqueous solution. CTAB micelles formed in water and minimized the distance between the donors and acceptors significantly by electrostatic interactions, improving FRET efficiency. Several factors that affected the fluorescence spectra of the FRET system were investigated. The prepared FRET system was feasible as an effective fluorescent probe to detect Hg(II) in aqueous solution. At pH7.0, a linear relationship between the quenched fluorescence intensity of orange-emitting acceptors (QDs(A) ) and Hg(II) concentration was acquired in the range 5-250nmol/L with a detection limit of 1.95nmol/L. The developed method showed excellent analytical performance for Hg(II) with high sensitivity and acceptable selectivity, reproducibility and stability. This finding indicated that the method has a promising potential application for environmental monitoring. This study demonstrated the great promise of QDs for expedient, low-cost and high-sensitivity detection of Hg(II).

Lignosulfonate: A Convenient Fluorescence Resonance Energy Transfer Platform for the Construction of a Ratiometric Fluorescence pH-Sensing Probe.

Lignin is a kind of natural fluorescent polymer material. However, the application based on the fluorescent property of lignin was rarely reported. Herein, a noncovalent lignin-based fluorescence resonance energy transfer (FRET) system was readily constructed by physical blending method with spirolactam rhodamine B (SRhB) and lignosulfonate (LS) as the acceptor and donor groups, respectively. The FRET behavior, self-assembly, and energy transfer mechanism of SRhB/LS composite were systematically studied. It was demonstrated that LS could be used as a convenient aptamer as energy donor to construct water-soluble ratiometric sensors because of its inherent property of intramicelle energy transfer cascades. Our results not only present a facile and general strategy for producing lignin-based functional material but also provide a fundamental understanding about lignin fluorescence to promote the functional and high-valued applications of lignin fluorescence characteristic.

Non-radiometric Cell-free Assay to Measure the Effect of Molecular Chaperones on AMP-activated Kinase Activity.

AMP-activated kinase (AMPK) is a trimeric protein holoenzyme with kinase activity. AMPK plays an important role in cellular metabolism and is thought to function as a fuel sensor within the cell, exerting kinase activity to activate energy-conserving pathways and simultaneously inhibit energy-consuming pathways. Traditional in vitro methods to measure AMPK activity to test potential agonists or antagonists utilize radiolabeled ATP with a peptide substrate. Although radiolabeling provides a high level of sensitivity, this approach is not ideal for medium to high-throughput screening, dose-response curves, or kinetic analyses. Our protocol utilizes Invitrogen's Z'-LYTE™ Kinase Assay Kit (Ser/Thr 23 Peptide) to measure changes in the enzymatic activity of AMPKɑ2β1γ1 in the presence of a molecular chaperone. The Z'-LYTE™ platform is based on Fluorescence Resonance Energy Transfer (FRET). The AMPK peptide substrate (S/T 23 peptide: MRPRKRQGSVRRRV) is a self-contained FRET system, using coumarin as the donor and fluorescein as the acceptor. When the peptide is phosphorylated, it is sensitive to cleavage by a site-specific protease. The cleavage of the phospho-peptide eliminates the FRET pair, and the ratiometric analysis of FRET is used as an indirect measure of AMPK kinase activity. This method does not require the use of radiolabeling or antibodies and is used in a multi-well format, with high reproducibility and throughput capabilities.

A fluorescence turn-on biosensor based on gold nanoclusters and aptamer for alpha fetoprotein detection

Alpha-fetoprotein (AFP), an important tumor marker, is widely used for the diagnosis of patients with germ cell tumors and hepatocellular carcinoma. Herein, a highly sensitive aptasensor for AFP detection was constructed based on fluorescence resonance energy transfer (FRET) between 5-carboxyfluorescein (FAM) and gold nanoclusters (AuNCs). AuNCs can effectively quench the fluorescence of FAM-AFP aptamer via strong coordination interactions. When AFP was introduced into the FAM-AFP aptamer-AuNCs FRET system, the AFP aptamer preferentially combined with AFP accompanied by conformational change, which greatly weakened the coordination interaction between the AFP aptamer and AuNCs. Thus, fluorescence recovery of FAM was observed. In the range of 10.0 -100.0 ng/mL, a linear relationship between the fluorescence recovery rate and the concentration of AFP was obtained with the detection limit of 6.631ng/mL. The highly sensitive FRET aptasensor with simple configuration showed promising prospect in detecting a variety of biomarkes.

An efficient fluorescence resonance energy transfer system from quantum dots to graphene oxide nano sheets: Application in a photoluminescence aptasensing probe for the sensitive detection of diazinon

In this paper a new aptasensor with high-sensitivity, and high-specificity for detection of diazinon, was discovered, based on fluorescence resonance energy transfer (FRET) between quantum dot (QD) as a donor and graphene oxide (GO) as an acceptor. l-cysteine capped CdS QDs/DF20 aptamer bioconjugates were successfully synthesized. GO was also attached to aptamers and photoluminescence quenching was obtained through FRET. By adding target diazinon to the bioconjugates containing GO, photoluminescence recovery was detected due to detachment of GO from the aptamer as a result of the difference in affinity for the aptamer. The detection limit of the biosensor was 0.13 nM and the linearity was maintained from 1.05 to 206 nM. Other pesticides and herbicides did not contribute to photoluminescence recovery due to lack of binding affinity for the aptamers, which demonstrates the selectivity of the biosensor. The results show the applicability of the aptasensor for monitoring diazinon in environmental and agricultural samples.

Achieving highly sensitive detection of Cu2+ based on AIE and FRET strategy in aqueous solution.

The aggregation-induced emission (AIE) luminogens are now showing strong potential in mimicking the energy donor of fluorescence resonance energy transfer (FRET) system. Herein, one highly efficient FRET system 1-NiR is successfully fabricated in aqueous solution based on an AIE active compound 1 and fluorescence dyes (Nile red (NiR)). 1 acts as the energy donor and NiR acts as the acceptor in the FRET system with the optimum concentrations ratio [1]/[NiR] = 100. Besides, the AIE(1) itself displays excellent selectivity for Cu2+ ions at 525 nm with the detection limit of 1.32 × 10-7 M. While through the FRET system of 1-NiR system, the detection limit of Cu2+ can be further decreased to 9.12 nM by monitoring the fluorescence at 630 nm. As a result, using an AIE probe to detect Cu2+ based on FRET mechanism is a promising strategy.

Drug screening assay based on the interaction of intact Keap1 and Nrf2 proteins in cancer cells.

The Nrf2-Keap1 interaction is the major regulatory pathway for cytoprotective responses against oxidative and electrophilic stresses. Keap1, a substrate protein of a Cul3-dependent E3 ubiquitin ligase complex, is a negative regulator of Nrf2. The use of chemicals to regulate the interaction between Keap1 and Nrf2 has been proposed as a strategy for the chemoprevention of degenerative diseases and cancers. The interactions between Keap1 and Nrf2 in vitro and in vivo were investigated using fluorescence resonance energy transfer (FRET) and bimolecular fluorescence complementation (BiFC) strategies in our study. Nrf2 with its N-terminal fused to eGFP and Keap1 with its C-terminal fused to mCherry were expressed and purified in vitro. When purified eGFP-Nrf2 and Keap1-mChrry proteins were mixed together, a strong FRET signal could be detected, indicating an efficient energy transfer from eGFP to mCherry. Moreover, the FRET was detected in vivo using confocal microscopy in colon cancer HCT-116 cells that were co-transfected with eGFP-Nrf2 and Keap1-mCherry. Finally, using an eGFP BiFC approach, the Keap1-Nrf2 interaction was also detected in MCF7 cells by transfecting eGFP N-terminal fused to Nrf2 (eN158-Nrf2) and eGFP C-terminal fused to Keap1 (eC159-Keap1). Using the BiFC and FRET systems, we demonstrated that the prototypical Nrf2-activiting compound tBHQ and the antitumor drug F-dUrd might interfere with the intracellular interaction between Keap1 and Nrf2 whereas the 5-Fu have little role in activating the protective response of Nrf2 pathway in cancer cells. By analyzing the perturbation of the energy transfer between the donor and acceptor fluorophores and the bimolecular fluorescence complementation of eGFP, we can screen potential inhibitors for the interaction between Keap1 and Nrf2.

The nature of photoinduced intermolecular charger transfer in fluorescence resonance energy transfer.

In this paper, we report time resolved fluorescence resonance energy transfer (FRET) using femtosecond ultrafast transient absorption spectroscopy. The lifetimes of FRET are strongly dependent on the molecular concentration and ratio of donor and acceptor. Also, in the FRET, photoinduced intermolecular charge transfer (PICT) is also investigated theoretically. The driving force for PICT in FRET system equals the reorganization energy, which gives barrier-less charge transfer according to Marcus theory. The rates of PICT in the FRET system can be estimated with our simplified Marcus equation. Our results of PICT in FRET system provide a new efficient way for the charge transfer in donor-acceptor system.

A Ratiometric Two-Photon Fluorescent Cysteine Probe with Well-Resolved Dual Emissions Based on Intramolecular Charge Transfer-Mediated Two-Photon-FRET Integration Mechanism.

The development of an efficient ratiometric two-photon fluorescence imaging probe is crucial for in situ monitoring of biothiol cysteine (Cys) in biosystems, but the current reported intramolecular charge transfer (ICT)-based one suffers from serious overlap between the shifted emission bands. To address this issue, we herein for the first time constructed an ICT-mediated two-photon excited fluorescence resonance energy transfer (TP-FRET) system consisting of a two-photon fluorogen benzo[ h]chromene and a Cys-responsive benzoxadiazole-analogue dye. Different from a previous mechanism that utilized single two-photon fluorogen to acquire a ratiometric signal, ICT was used to switch on the TP-FRET process of the energy transfer dyad by eliciting an absorption shift of benzoxadiazole with Cys to modulate the spectral overlap level between benzo[ h]chromene emission and benzoxadiazole absorption, resulting in two well-separated emission signal changes with large emission wavelength shift (120 nm), fixed two-photon excitation maximum (750 nm), and significant variation in fluorescence ratio (over 36-fold). Therefore, it can be successfully employed to ratiometrically visualize Cys in HeLa cells and liver tissues. Importantly, this new ICT-mediated TP-FRET integration mechanism would be convenient for designing ratiometric two-photon fluorescent probes with two well-resolved emission spectra suitable for high resolution two-photon fluorescence bioimaging.

The DNA Duplex as an Aqueous One-Dimensional Soft Crystal Scaffold for Photochemistry

Abstract In this account, we demonstrate that DNA duplex is an ideal scaffold for photochemistry, particularly for comparison of photochemical theory with experiments. The well-defined structure of a DNA duplex can be regarded as an aqueous one-dimensional soft crystal composed of a chromophore-like base-pair assembly. When any base pair in the duplex is replaced with a chromophore, orientation, distance, and association number of chromophores can be precisely controlled. We have developed a new methodology for introduction of chromophores into DNA duplexes using d-threoninol. By using the DNA duplex as a scaffold, experiments on exciton interactions of chromophore assemblies can be compared with molecular exciton theory. A fluorescent resonance energy transfer (FRET) system was also constructed by introducing donor pyrene and acceptor perylene into the DNA duplex using d-threoninol monomers. Using this system, we demonstrated orientation-dependent FRET. We found that theories on both exciton interaction and FRET qualitatively coincide with experimental data and revealed the limitation of the point-dipole approximation. We also evaluated the intrinsic quantum yield of photodimerization of stilbene derivatives by suppressing a side reaction. We propose that there is a correlation of quantum yield of photodimerization with the energy gap of HOMO or LUMO, a hypothesis that deserves theoretical investigation.

A dual-functional probe for quantification and imaging of intracellular telomerase

The analysis of telomerase is of great significance for the study and diagnosis of cancer. In this work, a dual-functional Au NP probe for the quantification and imaging of intracellular telomerase was prepared. The probe contained an elemental tag (1,4,7,10-tetraazacyclododecane-1,4,7-tris-acetic acid (Tb-DOTA)) for accurate quantification using inductively coupled plasma mass spectrometry (ICP-MS) and a fluorescent tag (FAM) which constructed the fluorescence resonance energy transfer (FRET) system with Au NPs for fluorescence imaging. After the probe entered into the Hela cells, the telomerase primer was extended with TTAGGG repeats in the presence of the intracellular telomerase, resulting in turning on the fluorescence of FAM and releasing the Tb-DOTA. Thus, the intracellular telomerase could be in situ visualized by the released FAM with fluorescence imaging and quantified by the released Tb-DOTA with ICP-MS detection. The average amount of telomerase in one Hela cell was quantified to be 3.1 × 10−11 IU. The proposed method was demonstrated with good potential in the study of telomerase and the cancer diagnosis, providing a new strategy for the analysis of intracellular biomarkers by ICP-MS.

Amplified fluorescence resonance energy transfer sensing of prostate specific antigen based on aggregation of CdTe QDs/antibody and aptamer decorated of AuNPs-PAMAM dendrimer

In this work, a simple and sensitive approach for detection of prostate-specific antigen (PSA) based on the fluorescence resonance energy transfer (FRET) from cadmium telluride (CdTe) quantum dots (QDs) to gold nanoparticles (AuNPs)-polyamidoamine (PAMAM) is described. In this FRET system, CdTe QD modified with antibody (Ab) acts as donor and aptamer decorated PAMAM-AuNPs is used as acceptor in the QDs-Ab/AuNPs-PAMAM/aptamer system. Detection of PSA is achieved by monitoring immunocomplex between PSA and aptamer, while the fluorescence intensity of CdTe QDs decreased during PSA addition. A linear relationship between the concentrations of the PSA antigen and the fluorescence-quenching intensities is obtained when QDs is closed to the AuNPs surface as a result of sandwich structure of QDs-Ab/PSA/AuNPs-PAMA/aptamer. Under the optimized conditions, the linear range of PSA antigen is extended from 0.01 ng mL−1 to 100 ng mL−1 with limit of detection (LOD) of 1 pg mL−1. A superior sensitivity is achieved due to the superior fluorescence properties of CdTe QDs along with highly loading ability of dendrimer and high selectivity of sensor toward PSA displayed due to more specificity of aptamer. The presented assay was further applied for PSA detection in human serum samples with satisfactory results. Due to high selectivity, simplicity and sensitivity of the proposed strategy, it could be developed for sensing of other targets by varying the recognition probes.