Classical Biosensors Research Articles

Genetic circuits in microbial biosensors for heavy metal detection in soil and water

With the rapid population growth, the world is witnessing an ever-increasing demand for energy and natural resources. Consequently, soil, air, and water are polluted with diverse pollutants, including heavy metals (HM). The detection of heavy metals is necessary to remediate them, which is achieved with biosensors. Initially, these HM were detected using atomic absorption spectroscopy (AAS), emission spectroscopy, mass spectrometry, gas chromatography etc., but these were costly and time consuming which further paved a way for microbe-based biosensors. The development of genetic circuits for microbe-based biosensors has become more popular in recent years for heavy metal detection. In this review, we have especially discussed the various types of genetic circuits such as toggle switches, logic gates, and amplification modules used in these biosensors as they are used to enhance sensitivity and specificity. Genetic circuits also allow for rapid and multiple analyte detection at the same time. The use of microbial biosensors for the detection of HM in the soil as well as the water is also described below. Although with a higher success rate than classical biosensors, these microbial biosensors still have some drawbacks like bioavailability and size of the analyte which are needed to be addressed.

A Scalable CMOS Molecular Electronics Chip for Single-Molecule Biosensing.

This work reports the first CMOS molecular electronics chip. It is configured as a biosensor, where the primary sensing element is a single molecule "molecular wire" consisting of a ∼100 GΩ, 25 nm long alpha-helical peptide integrated into a current monitoring circuit. The engineered peptide contains a central conjugation site for attachment of various probe molecules, such as DNA, proteins, enzymes, or antibodies, which program the biosensor to detect interactions with a specific target molecule. The current through the molecular wire under a dc applied voltage is monitored with millisecond temporal resolution. The detected signals are millisecond-scale, picoampere current pulses generated by each transient probe-target molecular interaction. Implemented in a 0.18 μm CMOS technology, 16k sensors are arrayed with a 20 μm pitch and read out at a 1 kHz frame rate. The resulting biosensor chip provides direct, real-time observation of the single-molecule interaction kinetics, unlike classical biosensors that measure ensemble averages of such events. This molecular electronics chip provides a platform for putting molecular biosensing "on-chip" to bring the power of semiconductor chips to diverse applications in biological research, diagnostics, sequencing, proteomics, drug discovery, and environmental monitoring.

Minimalistic design and rapid-fabrication single-mode fiber biosensors: Review and perspectives

Optical fiber biosensors are receiving significant interest, as they allow real-time, low-limit, and high precision detection of biological analytes such as biomarkers, proteins, and small cells. Classical biosensor designs include gratings, interferometers, and plasmonic structures; however, these systems require a complex design, hard to manufacture on high volumes. Recently, several biosensors having a minimalistic design, rapid and suitable to be automated, have been proposed; the key asset is the scalability potential, in view of fabricating numerous devices for disposable use or for large parallel immunoassays. This new sensor class features reflector-less sensors, shallow tapers, and ball resonators; a pseudo-random spectrum characterizes all these biosensors. In this paper, we provide a bird-eye view of these minimalistic designs and a perspective of their use in advanced biosensing applications.

Cellular Biosensors with Engineered Genetic Circuits.

An increasing interest in building novel biological devices with designed cellular functionalities has triggered the search of innovative tools for biocomputation. Utilizing the tools of synthetic biology, numerous genetic circuits have been implemented such as engineered logic operation in analog and digital circuits. Whole cell biosensors are widely used biological devices that employ several biocomputation tools to program cells for desired functions. Up to the present date, a wide range of whole-cell biosensors have been designed and implemented for disease theranostics, biomedical applications, and environmental monitoring. In this review, we investigated the recent developments in biocomputation tools such as analog, digital, and mix circuits, logic gates, switches, and state machines. Additionally, we stated the novel applications of biological devices with computing functionalities for diagnosis and therapy of various diseases such as infections, cancer, or metabolic diseases, as well as the detection of environmental pollutants such as heavy metals or organic toxic compounds. Current whole-cell biosensors are innovative alternatives to classical biosensors; however, there is still a need to advance decision making capabilities by developing novel biocomputing devices.

Developing enhanced magnetoimmunosensors based on low-cost screen-printed electrode devices

Abstract Electrochemical magnetoimmunosensors combine a number of issues that guarantee extremely high performance and also compatibility with the study of complex sample matrices. First, analyte immunocapture exploits the high affinity and specificity of antibodies. Second, magnetic particles (MP) provide faster and more efficient immunocapture than binding on two-dimensional structures, separation from nontarget sample components, and concentration of the target analyte. Finally, electrochemical detection supplies sensitivity and fast signal generation using robust and potentially miniaturized measurement equipment and transducers. On the contrary, MP handling is slightly more complex for end-users and more difficult to integrate in point-of-care devices than the manipulation of a classical biosensor. Attempts have been made to automate immunomagnetic binding, and the first robotized systems and platforms for the fluorescent and spectrophotometric detection of magnetoimmunoassays have already reached the market. Among the different types of electrodes available, screen-printed electrodes (SPE) stand out because of their low production cost and yet acceptable performance and interdevice reproducibility, which make them an excellent choice for analytical applications. In addition, each SPE entails a whole electrochemical cell stamped on a planar physical substrate, which makes it possible detection in small volumes and is especially favorable for the magnetic confinement of MP and the integration of microfluidic structures. In this article, we discuss the advantages obtained by using SPE and MP for the production of electrochemical magnetoimmunosensors and the clues for the successful development of such devices. We then revise some of the most outstanding works published in the literature.

ChemInform Abstract: Nanoparticles for Use in Enzyme Assays

AbstractReview: [55 refs.

Nanoparticles for Use in Enzyme Assays.

Nanoparticles (NPs) have created new ways to enhance the performance of classical biosensors in analytical sciences. NPs with unprecedented physiochemical properties can serve both as excellent carriers of bioreceptors and as signal enhancers, leading to improved assay platforms with high sensitivity and selectivity. Because enzymes play central roles in many cellular functions, specific and precise assays of their functions are of great significance in medical science and biotechnology. Here we review recent advances in NP-based biosensors and their use in enzyme assays. With fast and specific responses to enzyme-mediated reactions, NPs transduce and amplify the initial responses into various types of signals, such as electrochemical, optical and magnetic ones. Translation of their potential should lead to functionalized NPs finding wide applications in diagnostics, drug development and biotechnology.

Evanescent-field-induced Raman scattering for bio-friendly fingerprinting at sub-cellular dimension

Evanescent field induced chemical imaging concept has been realized in analytical platform based on the µ-tip-enhanced Raman scattering spectroscopy (µ-TERS). The technique aimed to minimize thermal decomposition of dried biological sample as the result of huge concentration of optical field near the tip by increasing the size of an aperture-less “excitation source”. µ-TERS technique is similar to classical biosensor systems based on propagating surface plasmon resonance phenomenon but with sensitive elements a few micrometers in size that can be targeted to the area of interest. The utility of the concept is exemplified by the analysis of dried single cell envelope of genetically modified Saccharomyces cerevisiae yeast cells, which do not have any heat-removing pathways, by water as in the case of the living cell. Practical excitation conditions effective for µ-TERS Raman observation of single layer dried biological samples without photodamage-related spectral distortion have been determined – the allowable limit is above 30s at 13µW/µm2. Finally, potential of µ-TERS spectroscopy as new bio-friendly instrumental platform for chemical fingerprinting and analytical characterization of buried nanoscale features is discussed.

Utilizing insect behavior in chemical detection by a behavioral biosensor

Traditionally, biosensors have been defined as consisting of two parts; a biological part, which is used to detect chemical or physical changes in the environment, and a corresponding electronic component, which tranduces the signal into an electronically readable format. Biosensors are used for detection of volatile compounds often at a level of sensitivity unattainable by traditional analytical techniques. Classical biosensors and traditional analytical techniques do not allow an ecological context to be imparted to the volatile compound/s under investigation. Therefore, we propose the use of behavioral biosensors, in which a whole organism is utilized for the analysis of chemical stimuli. In this case, the organism detects a chemical or physical change and demonstrates this detection through modifications of its behavior; it is the organism's behavior itself that defines the biosensor. In this review, we evaluate the use and future prospects of behavioral biosensors, with a particular focus on parasitic wasps.

Theoretical detection limits of magnetic biobarcode sensors and the phase space of nanobiosensing

A scaling theory of the sub atto-molar (aM) detection limits of magnetic particle (MP) based biosensors (e.g., bio-barcode assays) is developed and discussed. Despite the dramatic differences of sensing protocols and detection limits, the MP-based sensors can be interpreted within the same theoretical framework as any other classical biosensor (e.g., nanowire sensors), except that these sensors are differentiated by the geometry of diffusion and the probe (ρ(MP))/target (ρ(T)) density ratio. Our model predicts two regimes for biomolecule detection: For classical biosensors with ρ(MP) ≤ ρ(T), the response time t(s) proportional to 1/ρ(T); while for MP-based biosensors with ρ(MP) > ρ(T), t(s) proportional to 1/ρ(MP). The theory (i) explains the performance improvement of MP-sensors by ρ(MP)/ρ(T) (order of 10(3)-10(6)), broadly validating the sub-aM detection limits reported in literature, (ii) offers intuitive interpretation for the counter-intuitive ρ(T)-independence of detection time in MP-sensors, (iii) shows that statistical fluctuations should reduce with ρ(T) for MP sensors, and (iv) offers obvious routes to sensitivity improvement of classical sensors.

Point-of-care biosensor systems for cancer diagnostics/prognostics

With the growing number of fatalities resulting from the 100 or so cancer-related diseases, new enabling tools are required to provide extensive molecular profiles of patients to guide the clinician in making viable diagnosis and prognosis. Unfortunately with cancer-related diseases, there is not one molecular marker that can provide sufficient information to assist the clinician in making effective prognoses or even diagnoses. Indeed, large panels of markers must typically be evaluated that cut across several different classes (mutations in certain gene fragments--DNA; over/under-expression of gene activity as monitored by messenger RNAs; the amount of proteins present in serum or circulating tumor cells). The classical biosensor format (dipstick approach for monitoring the presence of a single element) is viewed as a valuable tool in many bioassays, but possesses numerous limitations in cancer due primarily to the single element nature of these sensing platforms. As such, if biosensors are to become valuable tools in the arsenal of the clinician to manage cancer patients, new formats are required. This review seeks to provide an overview of the current thinking on molecular profiling for diagnosis and prognosis of cancers and also, provide insight into the current state-of-the-art in the biosensor field and new strategies that must be considered to bring this important technology into the cancer field.

Electrochemical biosensing based on universal affinity biocomposite platforms

Rigid conducting biocomposites are versatile and effective transducing materials for the construction of a wide range of amperometric biosensors such as immunosensors, genosensors and enzymosensors, particularly if the transducer is bulk-modified with universal affinity biomolecules. The strept(avidin)-graphite-epoxy biocomposite could be considered as an universal immobilization platform whereon biotinylated DNAs, oligonucleotides, enzymes or antibodies can be captured by means of the highly affinity (strept)avidin-biotin reaction. Universal affinity biocomposite-based biosensors offer many potential advantages compared to more traditional electrochemical biosensors commonly based on a biologically surface-modified transducer. The integration of many materials into one matrix is their main advantage. As biological bulk-modified materials, the conducting biocomposites act not only as transducers, but also as reservoir for the biomaterial. After its use, the electrode surface can be renewed by a simple polishing procedure, establishing a clear advantage of these approaches relative to classical biosensors and other common biological assays. Moreover, the same material is useful for the analysis of many molecules whose determinations are based on genetic, enzymatic or immunological reactions. The different strategies for electrochemical genosensing, immunosensing and enzymosensing, all of them being dependent on the presence of a redox enzyme marker for the generation of the electrochemical signal, based on this universal affinity biocomposite platform are all presented and discussed.