A novel highly sensitive compilation-detachment fluorescence sensing strategy based on RNA-cleavage DNAzyme for MDA-MB-231 breast cancer biomarker determination.

Herein, we designed a novel and highly sensitive fluorescence multicomponent detachable platform for MDA-MB-231 breast cancer cell detection as a model. The RNA cleavage DNAzyme was used as a central operator of the multicomponent probe through which compilation and induced detachment of probe was done. During the compilation step, the dsDNA-Sybr green 1 complexes on gold nanoparticles (GNP@dsDNA@SG1) were assembled. The intercalated Sybr green in the DNA structure has been used as an amplified signal generator on one site of DNAzyme and magnetic nanoparticles (MNP) act as a biological carrier and probe collector on the opposite side. The enzyme activator co-factor (MDA-MB-231 cell cytoplasmic protein) provokes the activation of the catalytic core of enzyme sequence in the DNAzyme molecule, followed by cleavage reaction in the substrate sequence and releasing GNP@ dsDNA@SG1 into the solution. The results indicate that the Sybr green emission fluorescence (520 nm) increases with the increment of MDA-MB-231 protein concentration in the linear dynamic range of 8.10 × 10-2 to 1.95 ng ml-1 (0.77 × 10-3-0.019 cell ml-1) with a detection limit (LOD) of 1/72 × 10-2 pg ml-1 under optimal conditions. The proposed immunosensor has great potential in developing ultrasensitive and rapid diagnostic platforms.

Induced ultrasensitive electrochemical biosensor for target MDA-MB-231 cell cytoplasmic protein detection based on RNA-cleavage DNAzyme catalytic reaction.

Herein, we implemented RNA-cleaving DNAzymes specific for the endogenous protein of breast cancer cells (MDA-MB -231) and programmed for electrochemical detection. Thionine-modified gold nanoparticles and modified magnetic nanoparticles are attached to the two ends of the DNAzyme molecule. The prepared probe is pulled to the surface of the electrode with the help of a magnetic field, and the signal caused by the electrochemical activity of thionine is observed on the surface of the electrode. The presence of a covalent gold nanoparticle-thionine hybrid as a highly electroactive/enhanced electrochemical label ensures a strong detection signal. After addition of the enzyme activator cofactor (MDA-MB -231 cytoplasmic cell protein), it reacts with the catalytic core of the enzyme sequence in the DNAzyme molecule and triggers the cleavage reaction in the substrate sequence of the DNAzyme molecule. During this process, the gold nanoparticle-thionine labels are detached from the probe and released into the solution. Inductive removal of gold nanoparticles leads to a decrease in the current related to the reduction of thionine on the electrode surface. The results show that this biosensor can detect this protein marker in the linear range of (1.0E-06 to 1.0E+01) pg/ml, with a detection limit (1.0129E-07 pg/ml), using differential pulse voltammetry as a measuring technique. As well as, electrochemical impedance spectroscopy (EIS).

Exploring the Role of 2D-Graphdiyne as a Charge Carrier Layer in Field-Effect Transistors for Non-Covalent Biological Immobilization against Human Diseases.

Graphdiyne's (GDY's) outstanding features have made it a novel 2D nanomaterial and a great candidate for electronic gadgets and optoelectronic devices, and it has opened new opportunities for the development of highly sensitive electronic and optical detection methods as well. Here, we testified a non-covalent grafting strategy in which GDY serves as a charge carrier layer and a bioaffinity substrate to immobilize biological receptors on GDY-based field-effect transistor (FET) devices. Firm non-covalent anchoring of biological molecules via pyrene groups and electrostatic interactions in addition to preserved electrical properties of GDY endows it with features of an ultrasensitive and stable detection mechanism. With emerging new forms and extending the subtypes of the already existing fatal diseases, genetic and biological knowledge demands more details. In this regard, we constructed simple yet efficient platforms using GDY-based FET devices in order to detect different kinds of biological molecules that threaten human health. The resulted data showed that the proposed non-covalent bioaffinity assays in GDY-based FET devices could be considered reliable strategies for novel label-free biosensing platforms, which still reach a high on/off ratio of over 104. The limits of detection of the FET devices to detect DNA strands, the CA19-9 antigen, microRNA-155, the CA15-3 antigen, and the COVID-19 antigen were 0.2 aM, 0.04 pU mL-1, 0.11 aM, 0.043 pU mL-1, and 0.003 fg mL-1, respectively, in the linear ranges of 1 aM to 1 pM, 1 pU mL-1 to 0.1 µU mL-1, 1 aM to 1 pM, 1 pU mL-1 to 10 µU mL-1, and 1 fg mL-1 to 10 ng mL-1, respectively. Finally, the extraordinary performance of these label-free FET biosensors with low detection limits, high sensitivity and selectivity, capable of being miniaturized, and implantability for in vivo analysis makes them a great candidate in disease diagnostics and point-of-care testing.

Ultrasensitive fluorescence immunosensor based on mesoporous silica and magnetic nanoparticles: Capture and release strategy.

Herein we designed a novel, highly sensitive, simple and amplified fluorescence immunosensing strategy for hepatitis B virus surface antigen (HBV surface antigen) (HBsAg) as a model based on the construction of a sandwich type probe. The operation mechanism of this immunosensing strategy is implemented by capturing and then stimulation-based-releasing of entrapped dye in the fluorescent capsules. The proposed probe is made by the Fe3O4 magnetic nanoparticle (Fe3O4 MNP) as a probe collector site and the Rhodamine B loaded-mesoporous silica nanoparticle (MSN-Rh.B) as a fluorescent mesoporous capsule and signal amplifier site. Such a methodology is benefited, from the advantages of the high ability of MSNs to be used as a scaffold for efficient dye encapsulation and the magnetic nanoparticles as efficient biological carriers. Under optimal conditions, the fluorescence signal (The fluorescence of solutions was measured using a quartz fluorescence cell (PMT voltage:720, Ex wavelegth:540, Em wavelength:568, All measurements were carried out at room temperature) increased with the increment of HBsAg concentration in the linear dynamic range of 6.1 ag/ml to 0.012 ng/ml with a detection limit (LOD) of 5.7 ag/ml. The relative standard deviation, measured between the resulting fluorescence peaks was obtained by 6.0%.

A Novel Immunosensing Method Based on the Capture and Enzymatic Release of Sandwich-Type Covalently Conjugated Thionine-Gold Nanoparticles as a New Fluorescence Label Used for Ultrasensitive Detection of Hepatitis B Virus Surface Antigen.

A novel ultrasensitive and simple amplified immunosensing strategy is designed based on a surface-enhanced fluorescence (SEF) nanohybrid made from covalently conjugated thionine-gold nanoparticles (GNP-Th), as a novel amplified fluorescence label, and magnetic nanoparticles (MNPs), as a biological carrier, used for hepatitis B virus surface antigen (HBsAg) detection. This immunosensing strategy operates on the basis of the capture and then release of the amplified fluorescence label. Capturing of the antiHBs-antibody (Ab)-modified GNP-thionine hybrid (GNP-Th-Ab) is carried out through the formation of a two-dimensional (sandwich) probe between this amplified label and antiHBs-antibody-modified magnetic nanoparticles (MNP-Ab), in the presence of a target antigen and using an external magnetic force. Afterward, releasing of the captured fluorescence label is performed using a protease enzyme (pepsin) by a digestion mechanism of grafted antibodies on the GNP-thionine hybrid. As a result of antibody digestion, the amplified fluorescent hybrids (labels) are released into the solution. To understand the mechanism of enhanced fluorescence, the nature of the interaction between thionine and gold nanoparticles is studied using the B3LYP density functional method. In such a methodology, several new mechanisms and structures are used simultaneously, including a SEF-based metal nanoparticle-organic dye hybrid, dual signal amplification in a two-dimensional probe between the GNP-thionine hybrid and MNPs, and a novel releasing method using protease enzymes. These factors improve the sensitivity and speed, along with the simplicity of the procedure. Under optimal conditions, the fluorescence signal increases with the increment of HBs antigen concentration in the linear dynamic range of 4.6 × 10-9 to 0.012 ng/mL with a detection limit (LOD) of 4.6 × 10-9 ng/mL. The proposed immunosensor has great potential in developing ultrasensitive and rapid diagnostic platforms.