A Ready-to-Use Metal-Supported Bilayer Lipid Membrane Biosensor for the Detection of Phenol in Water.

This work presents a novel metal-supported bilayer lipid membrane (BLM) biosensor built on tyrosinase to quantitate phenol. The detection strategy is based on the enzyme-analyte initial association and not the commonly adopted monitoring of the redox cascade reactions; such an approach has not been proposed in the literature to date and offers many advantages for environmental monitoring with regard to sensitivity, selectivity, reliability and assay simplicity. The phenol sensor developed herein showed good analytical and operational characteristics: the detection limit (signal-to-noise ratio = 3) was 1.24 pg/mL and the sensitivity was 33.45 nA per pg/mL phenol concentration. The shelf life of the tyrosinase sensor was 12 h and the lifetime (in consecutive assays) was 8 h. The sensor was reversible with bathing at pH 8.5 and could be used for eight assay runs in consecutive assays. The validation in real water samples showed that the sensor could reliably detect 2.5 ppb phenol in tap and river water and 6.1 ppb phenol in lake water, without sample pretreatment. The prospects and applicability of the proposed biosensor and the underlying technology are also discussed.

The Application of Lipid Membranes in Biosensing.

The exploitation of lipid membranes in biosensors has provided the ability to reconstitute a considerable part of their functionality to detect trace of food toxicants and environmental pollutants. This paper reviews recent progress in biosensor technologies based on lipid membranes suitable for food quality monitoring and environmental applications. Numerous biosensing applications based on lipid membrane biosensors are presented, putting emphasis on novel systems, new sensing techniques, and nanotechnology-based transduction schemes. The range of analytes that can be currently using these lipid film devices that can be detected include, insecticides, pesticides, herbicides, metals, toxins, antibiotics, microorganisms, hormones, dioxins, etc. Technology limitations and future prospects are discussed, focused on the evaluation/validation and eventually commercialization of the proposed lipid membrane-based biosensors.

Application of Biosensors Based on Lipid Membranes for the Rapid Detection of Toxins.

Lipid assemblies in the form of two dimensional films have been used extensively as biosensing platforms. These films exhibit certain similarities with cell membranes, thus providing a suitable means for the immobilization of proteinaceous moieties and, further, a number of intrinsic signal amplification mechanisms. Their implementation in the detection of toxins yielded reliable and fast detectors for in field analyses of environmental and clinical samples. Some examples are presented herein, including aflatoxin and cholera toxin detection. The conditions and parameters that determine the analytical specifications of the lipid membrane sensors are discussed, advantages and technology bottlenecks are reviewed, and possible further developments are highlighted.

Novel Biosensors for the Rapid Detection of Toxicants in Foods.

The modern environmental and food analysis requires sensitive, accurate, and rapid methods. The growing field of biosensors represents an answer to this demand. Unfortunately, most biosensor systems have been tested only on distilled water or buffered solutions, although applications to real samples are increasingly appearing in recent years. In this context, biosensors for potential food applications continue to show advances in areas such as genetic modification of enzymes and microorganisms, improvement of recognition element immobilization, and sensor interfaces. This chapter investigates the progress in the development of biosensors for the rapid detection of food toxicants for online applications. Recent progress in nanotechnology has produced affordable, mass-produced devices, and to integrate these into components and systems (including portable ones) for mass market applications for food toxicants monitoring. Sensing includes chemical and microbiological food toxicants, such as toxins, insecticides, pesticides, herbicides, microorganisms, bacteria, viruses and other microorganisms, phenolic compounds, allergens, genetically modified foods, hormones, dioxins, etc. Therefore, the state of the art of recent advances and future targets in the development of biosensors for food monitoring is summarized as follows: biosensors for food analysis will be highly sensitive, selective, rapidly responding, real time, massively parallel, with no or minimum sample preparation, and platform suited to portable and handheld nanosensors for the rapid detection of food toxicants for online uses even by nonskilled personnel.

Lipid Membrane Nanosensors for Environmental Monitoring: The Art, the Opportunities, and the Challenges.

The advent of nanotechnology has brought along new materials, techniques, and concepts, readily adaptable to lipid membrane-based biosensing. The transition from micro-sensors to nano-sensors is neither straightforward nor effortless, yet it leads to devices with superior analytical characteristics: ultra-low detectability, small sample volumes, better capabilities for integration, and more available bioelements and processes. Environmental monitoring remains a complicated field dealing with a large variety of pollutants, several decomposition products, or secondary chemicals produced ad hoc in the short- or medium term, many sub-systems affected variously, and many processes largely unknown. The new generation of lipid membranes, i.e., nanosensors, has the potential for developing monitors with site-specific analytical performance and operational stability, as well as analyte-tailored types of responses. This review presents the state-of-the art, the opportunities for niche applicability, and the challenges that lie ahead.

Artificial Lipid Membranes: Past, Present, and Future.

The multifaceted role of biological membranes prompted early the development of artificial lipid-based models with a primary view of reconstituting the natural functions in vitro so as to study and exploit chemoreception for sensor engineering. Over the years, a fair amount of knowledge on the artificial lipid membranes, as both, suspended or supported lipid films and liposomes, has been disseminated and has helped to diversify and expand initial scopes. Artificial lipid membranes can be constructed by several methods, stabilized by various means, functionalized in a variety of ways, experimented upon intensively, and broadly utilized in sensor development, drug testing, drug discovery or as molecular tools and research probes for elucidating the mechanics and the mechanisms of biological membranes. This paper reviews the state-of-the-art, discusses the diversity of applications, and presents future perspectives. The newly-introduced field of artificial cells further broadens the applicability of artificial membranes in studying the evolution of life.

Protein-Based Graphene Biosensors: Optimizing Artificial Chemoreception in Bilayer Lipid Membranes.

Proteinaceous moieties are critical elements in most detection systems, including biosensing platforms. Their potential is undoubtedly vast, yet many issues regarding their full exploitation remain unsolved. On the other hand, the biosensor formats with the higher marketability probabilities are enzyme in nature and electrochemical in concept. To no surprise, alternative materials for hosting catalysis within an electrode casing have received much attention lately to demonstrate a catalysis-coated device. Graphene and ZnO are presented as ideal materials to modify electrodes and biosensor platforms, especially in protein-based detection. Our group developed electrochemical sensors based on these nanomaterials for the sensitive detection of cholesterol using cholesterol oxidase incorporated in stabilized lipid films. A comparison between the two platforms is provided and discussed. In a broader sense, the not-so-remote prospect of quickly assembling a protein-based flexible biosensing detector to fulfill site-specific requirements is appealing to both university researchers and industry developers.

A methodological combined framework for roadmapping biosensor research: a fault tree analysis approach within a strategic technology evaluation frame.

Biosensor technology began in the 1960s to revolutionize instrumentation and measurement. Despite the glucose sensor market success that revolutionized medical diagnostics, and artificial pancreas promise currently the approval stage, the industry is reluctant to capitalize on other relevant university-produced knowledge and innovation. On the other hand, the scientific literature is extensive and persisting, while the number of university-hosted biosensor groups is growing. Considering the limited marketability of biosensors compared to the available research output, the biosensor field has been used by the present authors as a suitable paradigm for developing a methodological combined framework for "roadmapping" university research output in this discipline. This framework adopts the basic principles of the Analytic Hierarchy Process (AHP), replacing the lower level of technology alternatives with internal barriers (drawbacks, limitations, disadvantages), modeled through fault tree analysis (FTA) relying on fuzzy reasoning to count for uncertainty. The proposed methodology is validated retrospectively using ion selective field effect transistor (ISFET) - based biosensors as a case example, and then implemented prospectively membrane biosensors, putting an emphasis on the manufacturability issues. The analysis performed the trajectory of membrane platforms differently than the available market roadmaps that, considering the vast industrial experience in tailoring and handling crystallic forms, suggest the technology path of biomimetic and synthetic materials. The results presented herein indicate that future trajectories lie along with nanotechnology, and especially nanofabrication and nano-bioinformatics, and focused, more on the science-path, that is, on controlling the natural process of self-assembly and the thermodynamics of bioelement-lipid interaction. This retained the nature-derived sensitivity of the biosensor platform, pointing out the differences between the scope of academic research and the market viewpoint.

Nanobodies as novel agents for disease diagnosis and therapy.

The discovery of naturally occurring, heavy-chain only antibodies in Camelidae, and their further development into small recombinant nanobodies, presents attractive alternatives in drug delivery and imaging. Easily expressed in microorganisms and amenable to engineering, nanobody derivatives are soluble, stable, versatile, and have unique refolding capacities, reduced aggregation tendencies, and high-target binding capabilities. This review outlines the current state of the art in nanobodies, focusing on their structural features and properties, production, technology, and the potential for modulating immune functions and for targeting tumors, toxins, and microbes.

Innovation in biotechnology: moving from academic research to product development--the case of biosensors.

The fast pace of technological change in the biotechnology industry and the market demands require continuous innovation, which, owing to the science base of the sector, derives from academic research through a transformation process that converts science-oriented knowledge to marketable products. There appear to be some inherent difficulties in transforming directly the knowledge output of academic research to industrial use. The purpose of this article is to examine certain transition mechanisms from monodisciplinary academic isolation (curiosity-driven and internal-worth innovation) to university-industry alliances (market-driven and public-worth innovation) through inter-organizational multidisciplinary collaboration and contextualize the analysis with the case of biosensors. While the majority of literature on the subject studies the channels of knowledge transfer as determinants of alliance success (transferor/transferee interactions), either from the university side (science base) or the industry side (market base), this article focuses on the transferable (technology base) and how it can be strategically modeled and managed by the industry to promote innovation. Based on the valuable lessons learnt from the biosensor paradigm, the authors argue that strategic industry choices deal primarily with the best stage/point to intersect and seize the university output, implanting the required element of marketability that will transform an idea to a viable application. The authors present a methodological approach for accelerating the knowledge transfer from the university to industry aiming at the effective transition of science to products through a business model reconfiguration.

A novel system for environmental monitoring through a cooperative/synergistic scheme between bioindicators and biosensors.

This paper addresses environmental monitoring through a robust dynamic integration between biomonitor and biosensor systems, a strategy that has not been attempted before. The two systems are conceptually interrelated and methodologically correlated to a cooperative/synergistic scheme (CSS) with a view to minimise uncertainty and monitoring costs and increase reliability of pollution control and abatement. The structures and operations of the biosensor component (in terms of sensitivity, device and method versatility, nature-mimicking physicochemical mechanisms, prospects and technological input) are such that they reinforce or promote the structures and operations of the natural component (in terms of bio-surveillance, impact assessment, environmental quality indexing, stress responses, metabolic pathways, etc.) and vice versa. The bioindicator ontology presented herein, including concepts, relations and controlled vocabulary aiming at establishing an integrated methodology for mapping/assessing negative environmental externalities, provides a useful tool for the design/development/implementation of an environmental network for the monitoring of a variety of pollutants over time and space and the assessment of environmental quality; the collection of the available information and its classification into taxonomic and partonomic relations allows the construction of a database that links pollutants with organisms' response, at a phenomenological and in-depth level, considering ecological parameters, relations and geomorphologic characteristics. As a result, a computer program has been designed/developed as a decision support system and has been successfully tested on a representative population of species indigenous to southern Greece. Significantly, a novel system in the form of a rational framework at the conceptual design level has been developed, that actually contributes towards achieving a cost-effective long-term monitoring program, with the flexibility to counter on-course any (anticipated or not) variations/modifications of the surveillance environment. This novel and pioneering approach will further offer a dynamic system utilised in (a) environmental impact studies and risk assessment (positive/analytic approach), (b) decision-making in the short-run (normative/tactic approach), and (c) policy-making in the long-run (normative/strategic approach). The proposed CSS, based on the integration of multiple data sources, can establish a local area network, incorporated into/expanding to a wide area network, thus offering the potential of better predictive ability and greater lead-time warning at alarm conditions than that provided by separate, stand-alone surveillance modalities.

A knowledge-based approach to environmental biomonitoring.

This paper presents the design, development and implementation of an integrated GIS-controlled knowledge-based system for environmental monitoring applications, utilizing indigenous flora for assessing quality. The system gathers and combines geographical, ecological, and physicochemical data of organisms' response to pollution within an intelligent computer program that (a) recognises groups of indigenous species suitable for long-term monitoring of a specific pollutant or a combination of pollutants, (b) estimates the ambient concentration of pollutant(s) from the population of the species comprising the bioindicator group and (c) provides biomonitoring capacity indices at national and international/transboundary levels. Significantly, a novel system in the form of a rational framework at the conceptual design level has been developed, that actually contributes towards achieving a cost-effective long-term biomonitoring program, with the flexibility to counter on-course any (anticipated or not) variations/modifications of the surveillance environment: the scheme assumes a robust dynamic cooperation between instrumental and biomonitoring systems, with a view to minimise uncertainty and monitoring costs and increase reliability of pollution control and abatement, aiming eventually at the shifting, partially or totally, from instrumental to natural monitoring. The proposed approach is presently implemented at pilot-scale for establishing a biomonitoring network at a large industrial area in Greece. The results obtained indicate that a cost-effective program can be only attained and maintained under a suitable financial/organizational scheme at the macro level, whereas the micro level viability strongly depends upon careful management of human resources and fixed assets.