Platform For Development Of Biosensors Research Articles

RNA reporter based CRISPR/Cas12a biosensing platform for sensitive detection of circulating tumor DNA.

CRISPR/Cas biotechnology provides an exceptional platform for biosensor development. To date, the reported CRISPR/Cas biosensing systems have shown extraordinary performance for nucleic acids, small molecules, small proteins and microorganism detection. The CRISPR/Cas12a biosensing system, as a typical example, has been well established and applied for both nucleic acids and non-nucleic acids target detection. However, all established CRISPR/Cas12a biosensing systems are based on DNA reporters, which potentially limits further application.In this study, we established an RNA reporter based CRISPR/Cas12a biosensing system. A basic biosensing system was evaluated, and the limit of detection was found to be 1 nM. Afterwards, we optimized this biosensing system using both temperature and chemical enhancers. The final optimal biosensing system (with DTT & 37°C) shows fluorescence signal increased by a factor of ~10 compared with the basic system. The optimal biosensing system was further applied for the detection of circulating tumor DNA (ctDNA), which shows over 4 orders of magnitude detection range from 1pM to 25 nM, with the limit of detection of 1pM. This RNA reporter based CRISPR/Cas12a biosensing system provides an effective platform for nucleic acids quantification.Clinical Relevance- This research provides a novel approach for ctDNA diagnostics, which is an attractive biomarker for noninvasive monitoring of tumor growth, response, and spread.

Block copolymer nanoparticles-based fluorescent sensor for ultrasensitive detection of tyrosinase activity and inhibitor

It is of great value to develop a simple, rapid, label-free and sensitive strategy for tyrosinase activity detection and corresponding inhibitor screening in both biomedical diagnosis and cosmetic industry. However, many recently reported fluorescent assays may suffer from instability to light irradiation, poor water solubility, and complex modification. Herein, a novel fluorescent biosensor strategy has been developed capable of sensitive, label-free and rapid screening of tyrosinase activity as well as corresponding inhibitors based on fluorescent block copolymer nanoparticles (BCNs). The reported BCNs were prepared by coprecipitation assay using the mixture of amphiphilic block copolymer and fluorescent conjugated polymer, which show many advantages including high brightness, excellent photostability and biocompatibility, enabling the biosensor with high sensitivity and good reproducibility in a single-step operation. The reported assay exhibited high sensitivity and selectivity with a detection limit of 1.5 ng/mL for tyrosinase (TRY) activity detection and hold potential in its inhibitor screening. The satisfying recoveries offered great potential for complicated sample analysis. Hence, this reported assay may provide a novel platform for biosensor development.

Development of cobalt oxide film modified electrode decorated with platinum nanoparticles as a biosensing platform for phenol

ABSTRACT Recently, cobalt oxides attract a special attention mainly due to their excellent electrocatalytic activity towards various compounds. This study concerns with electrochemical preparation of cobalt oxide modified glassy carbon electrodes decorated with platinum nanoparticle in the pursuit of its catalytic applications. The parameters affecting the deposition conditions have been evaluated by monitoring oxygen reduction reaction (ORR) in pH 5.0 acetate buffer. Under optimal conditions, the composite film electrode surface was utilised as a platform forbiosensor development. Polyphenol oxidase enzyme was immobilised onto the electrode surface for phenol detection, and the influence of solution and operational parameters have been investigated. The medium pH and the enzyme amount have been optimised as pH 7.0 and 0.3 mg, respectively. Chronoamperograms were recorded for phenol detection in micromolar levels.

H2O2 biosensor consisted of hemoglobin-DNA conjugate on nanoporous gold thin film electrode with electrochemical signal enhancement

In this research, we developed electrochemical biosensor which was composed of hemoglobin (Hb)-DNA conjugate on nanoporous gold thin film (NPGF) for hydrogen peroxide (H2O2) detection. For the first time, Hb and DNA was conjugated as a sensing platform for uniform orientation of Hb on electrode. The newly developed Hb-DNA conjugate was designed to prevent Hb from aggregation on electrode. DNA hybridization of Hb-DNA conjugate and complementary DNA (cDNA) on NPGF electrode induced uniformly assembled biosensor. Furthermore, NPGF electrode fabrication method was introduced to the increment of the surface area. To confirm the conjugation of Hb-DNA conjugate, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and ultraviolet–visible spectroscopy (UV–VIS) were used. Formation of the NPGF electrode was verified by scanning electron microscope (SEM). Atomic force microscopy (AFM) was operated for the confirmation of Hb-DNA immobilization on electrode. The electrochemical property of fabricated electrode was investigated by cyclic voltammetry (CV). Also, H2O2 sensing performance of fabricated electrode was investigated by amperometric i-t curve technique. This sensor showed a wide linear range from 0.00025 to 5.00 mM and a correlation coefficient of R2 = 0.9986. The detection limit was 250 nM. Proposed biosensor can be utilized as a sensing platform for development of biosensor.

Bacterial Surface Layer Proteins: A Promising Nano-Technological Tool for Bio-Sensing Applications

The phenomenal rise in the demand of biosensors accelerated their rapid development and immersive applications in the myriads of fields. The essential requirement of developing efficient bio-sensing platform is to find stable well organized interfacial architecture that can serve as an excellent matrix for binding and recognizing biomolecules. In this context, the enormous potential has been envisaged in surface layer proteins that represented themselves as most primitive and simplest self-assembled system with repetitive physicochemical properties for the molecular functionalization of surfaces and various interfaces. The prominence of S-layer proteins has been broadened by integrating genetic engineering approaches for the fine tuning of functional groups and protein domains in geometrically well-defined manner. The efficient and stable binding of various nanomaterials with S-layers in regular arrays has led to paradigmatic shift in their nano-biotechnological sensing applications. More recently, functional S-layer supported lipid membranes have been generated through covalent binding of lipid molecules either with native or recombinant S-layer proteins at nano-scale dimensions serving as “proof of concept” for the development of bio-sensing platform. Thus, in the light of benefits conferred by surface layer proteins for the development of highly efficient biosensors, an exciting path has been opened for broadening their translational applications in drug delivery, disease diagnosis, vaccines development, lab-on-chip devices etc. Therefore, this review intends to describe about the importance of surface layer proteins in the development of biosensors.

Mixture of PLA-PEG and biotinylated albumin enables immobilization of avidins on electrospun fibers.

The application of nanotechnology in biomedical field has enormous potential in basic and applied research. Micro or nanofibers produced by electrospinning technique offer excellent properties because of large specific surface area, high porosity, and ability to incorporate functional additives. Here we embedded biotinylated bovine serum albumin into polylactic acid (PLA)-polyethylene glycol (PEG) fibers, which enabled specific immobilization of fluorescently labelled avidin. An alkaline phosphatase enzyme was immobilized via biotin-streptavidin interaction on the hybrid nanofibers, demonstrating the suitability of the material for biosensing applications. These functional nanofibers provide a promising platform for development of biosensors and other biofunctional materials utilizing avidin-biotin as a generic and robust immobilization method. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 356-362, 2017.

Preparation and modification of carbon nanotubes electrodes by cold plasmas processes toward the preparation of amperometric biosensors

An electrochemical transducer based on vertically aligned carbon nanotubes (CNT) was prepared as a platform for biosensor development. Prior to enzyme immobilization, the CNT were treated using a microwave plasma system (CO 2 and N 2/H 2) in order to functionalize the CNT surface with oxygenated and aminated groups. The morphological aspect of the electrode surface was examined by SEM and its chemical structure was also elucidated by XPS analysis. It was found out that microwave plasma system (CO 2 and N 2/H 2) not only functionalizes the CNT but also permits to avoid the collapse phenomena retaining thus the alignment structure of the electrode surface. The electrochemical properties of the resulting new material based on CNT were carried out by cyclic voltammetry and were found suitable to develop high sensitive enzyme (HRP) biosensors operating on direct electron transfer process.