Femtomolar Detection of Dengue Virus DNA with Serotype Identification Ability.

Dengue surveillance trusts only on reverse transcription-polymerase chain reaction (RT-PCR) type methodologies for confirmation of dengue virus serotypes; however, its real time application is restricted due to the expensive, complicated, and time-consuming process. In search of a new sensing system, here, we have reported a two-way-detection method for Dengue virus (DENV) serotype identification along with DNA quantification by using a new class of nanocomposite of gold nanoparticles (AuNP) and nitrogen, sulfur codoped graphene quantum dots (N,S-GQDs). The N,S-GQDs@AuNP has been used for serotype detection via a simple fluorescence technique using four dye-combined probe DNAs which is further validated by confocal microscopy. The quantification of the DNA has been measured by the differential pulse voltammetric (DPV) technique using methyelene blue as a redox indicator. Results obtained in this study, clearly demonstrate that the N,S-GQDs@AuNP can efficiently detect the four serotypes of DENV individually in the concentration range of 10-14 to 10-6 M with the LOD of 9.4 fM. In addition, to confirm its applicability in long chained complex DNA system, the sensor was also applied to the clinically isolated DENV DNA and showed satisfactory performances for serotype identification as well as quantification. We hope this simple and reliable method can pave an avenue for the development of sensitive and robust sensing probes in biomedical applications.

Multifunctional GQDs-Concanavalin A@Fe3O4 nanocomposites for cancer cells detection and targeted drug delivery.

Multifunctional nanocomposites containing intrinsic property for serving as the sensing elements as well as targeted nanoconjugates are highly preferred in various therapeutic applications. In this work, nanocomposites of graphene quantum dots (GQDs) and Fe3O4 with conjugation of lectin protein, concanavalin A, to form GQD-ConA@Fe3O4 nanocomposites are developed for both detection of cancer cell and release of drugs to HeLa cells. The GQD-ConA@Fe3O4 nanocomposites deposited on Pt electrode can detect cancerous HeLa cells over normal endothelial cells with a dynamic linear range of 5 × 102 to 1 × 105 cells mL-1 with a detection limit of 273 cell mL-1. The GQD-ConA@Fe3O4 also can serve as nanocarriers for loading and delivering doxorubicin (Dox). The in vitro cell images show that the Dox concentration in HeLa cells is enhanced more than double in the presence of external magnetic field due to the incorporation of Fe3O4 in the nanocarrier. The cytotoxicity assay indicates that the susceptibility of cancerous HeLa cells to Dox is 13% higher than that of normal cells, confirming the selective role of ConA in nanocarriers. Results clearly indicate the GQD-ConA@Fe3O4 nanocomposites as a promising material for cancer cell detection and targeted Dox release toward HeLa cells which can serve as the multifunctional platform for novel cancer cell diagnostic and therapeutic applications.

N-Doped Graphene Quantum Dots-Decorated V2O5 Nanosheet for Fluorescence Turn Off-On Detection of Cysteine.

The development of a fast-response sensing technique for detection of cysteine can provide an analytical platform for prescreening of disease. Herein, we have developed a fluorescence turn off-on fluorescence sensing platform by combining nitrogen-doped graphene quantum dots (N-GQDs) with V2O5 nanosheets for the sensitive and selective detection of cysteine in human serum samples. V2O5 nanosheets with 2-4 layers are successfully synthesized via a simple and scalable liquid exfoliation method and then deposited with 2-8 nm of N-GQDs as the fluorescence turn off-on nanoprobe for effective detection of cysteine in human serum samples. The V2O5 nanosheets serve as both fluorescence quencher and cysteine recognizer in the sensing platform. The fluorescence intensity of N-GQDs with quantum yield of 0.34 can be quenched after attachment onto V2O5 nanosheets. The addition of cysteine triggers the reduction of V2O5 to V4+ as well as the release of N-GQDs within 4 min, resulting in the recovery of fluorescence intensity for the turn off-on detection of cysteine. The sensing platform exhibits a two-stage linear response to cysteine in the concentration range of 0.1-15 and 15-125 µM at pH 6.5, and the limit of detection is 50 nM. The fluorescence response of N-GQD@V2O5 exhibits high selectivity toward cysteine over other 22 electrolytes and biomolecules. Moreover, this promising platform is successfully applied in detection of cysteine in human serum samples with excellent recovery of (95 ± 3.8) - (108 ± 2.4)%. These results clearly demonstrate a newly developed redox reaction-based nanosensing platform using N-GQD@V2O5 nanocomposites as the sensing probe for cysteine-associated disease monitoring and diagnosis in biomedical applications, which can open an avenue for the development of high performance and robust sensing probes to detect organic metabolites.

Label-Free and Nondestructive Separation Technique for Isolation of Targeted DNA from DNA-Protein Mixture Using Magnetic Au-Fe3O4 Nanoprobes.

The interest in DNA-protein-based diagnostics has recently been growing enormously, which makes the separation process of DNA or protein from a cell extract extremely important. Unlike the traditional separation process, a novel approach is in demand which can nondestructively isolate the target biomolecules without sacrificing the other components in the mixture. In this study, we have demonstrated a new and simple separation technique by using well-established bifunctional Au-Fe3O4 nanocomposites as the separation nanoprobes to efficiently isolate the specifically targeted nanomolar concentrated DNA over 70% from its associate DNA-protein mixture in the presence of a magnetic field. The sensing accuracy of both as-separated DNA and protein are quantitatively examined by UV-vis spectroscopy, and then qualitatively validated by gel analysis. Results obtained in this study clearly demonstrated that this newly developed separation procedure not only provides the efficient separation for the targeted DNA but can also maintain the bioactivity of as-separated protein and DNA solutions. The superiority of this technique can open an avenue to establish a label-free and nondestructive platform for a wide variety of biomolecule separation applications.

Highly Sensitive and Selective Detection of Nanomolar Ferric Ions Using Dopamine Functionalized Graphene Quantum Dots.

The good stability, low cytotoxicity, and excellent photoluminescence property of graphene quantum dots (GQDs) make them an emerging class of promising materials in various application fields ranging from sensor to drug delivery. In the present work, the dopamine-functionalized GQDs (DA-GQDs) with stably bright blue fluorescence were successfully synthesized for low level Fe(3+) ions detection. The as-synthesized GQDs are uniform in size with narrow-distributed particle size of 4.5 ± 0.6 nm and high quantum yield of 10.2%. The amide linkage of GQDs with dopamine, confirmed by using XPS and FTIR spectra, results in the specific interaction between Fe(3+) and catechol moiety of dopamine at the interfaces for highly sensitive and selective detection of Fe(3+). A linear range of 20 nM to 2 µM with a detection limit of 7.6 nM is obtained for Fe(3+) detection by DA-GQDs. The selectivity of DA-GQDs sensing probe is significantly excellent in the presence of other interfering metal ions. In addition, the reaction mechanism for Fe(3+) detection based on the complexation and oxidation of dopamine has been proposed and validated. Results obtained in this study clearly demonstrate the superiority of surface functionalized GQDs to Fe(3+) detection, which can pave an avenue for the development of high performance and robust sensing probes for detection of metal ions and other organic metabolites in environmental and biomedical applications.