Computational design and clinical demonstration of a copper nanocluster based universal immunosensor for sensitive diagnostics.

Nanoparticle based sensors are good alternatives for non-enzymatic sensing applications due to their high stability, superior photoluminescence, biocompatibility and ease of fabrication, with the only disadvantage being the cost of the synthesis process (owing to the expensive precursors and infrastructure). For the first time, we report the design of an immunosensor employing streptavidin conjugated copper nanocluster, developed at a much lower cost compared to other nanomaterials like noble metal nanoparticles and quantum dots. Using in silico tools, we have tried to establish the dynamics of conjugation of nanocluster to the streptavidin protein, based on EDC-NHS coupling. The computational simulations have successfully explained the crucial role played by the components of the immunosensor leading to an efficient design capable of high sensitivity. In order to demonstrate the functioning of the Copper Nanocluster ImmunoSensor (CuNIS), HIV-1 p24 biomarker test was chosen as the model assay. The immunosensor was able to achieve an analytical limit of detection of 23.8 pg mL-1 for HIV-1 p24 with a linear dynamic range of 27-1000 pg mL-1. When tested with clinical plasma samples, CuNIS based p24 assay showed 100% specificity towards HIV-1 p24. With the capability of multiplexed detection and a cost of fabrication 100 times lower than that of the conventional metal nanoclusters, CuNIS has the potential to be an essential low-cost diagnostic tool in resource-limited settings.

Streptavidin-conjugated gold nanoclusters as ultrasensitive fluorescent sensors for early diagnosis of HIV infection.

We have engineered streptavidin-labeled fluorescent gold nanoclusters to develop a gold nanocluster immunoassay (GNCIA) for the early and sensitive detection of HIV infection. We performed computational simulations on the mechanism of interaction between the nanoclusters and the streptavidin protein via in silico studies and showed that gold nanoclusters enhance the binding to the protein, by enhancing interaction between the Au atoms and the specific active site residues, compared to other metal nanoclusters. We also evaluated the role of glutathione conjugation in binding to gold nanoclusters with streptavidin. As proof of concept, GNCIA achieved a sensitivity limit of detection of HIV-1 p24 antigen in clinical specimens of 5 pg/ml, with a detection range up to1000 pg/ml in a linear dose-dependent manner. GNCIA demonstrated a threefold higher sensitivity and specificity compared to enzyme-linked immunosorbent assay for the detection of HIV p24 antigen. The specificity of the immunoassay was 100% when tested with plasma samples negative for HIV-1 p24 antigen and positive for viruses such as hepatitis B virus, hepatitis C virus, and dengue. GNCIA could be developed into a universal labeling technology using the relevant capture and detector antibodies for the specific detection of antigens of various pathogens in the future.

Aqueous based reflux method for green synthesis of nanostructures: Application in CZTS synthesis.

The aqueous based reflux method useful for the green synthesis of nanostructures is described in detail. In this method, the parameters: the order of addition of precursors, the time of the reflux and the cooling rate should be optimized in order to obtain the desired phase and morphology of the nanostructures. The application of this method is discussed with reference to the synthesis of CZTS nanoparticles which have great potential as an absorber material in the photovoltaic devices. The highlights of this method are:•Simple.•Low cost.•Aqueous based.