Innovative biosensors for environment and health

The platform used for “Innovative Biosensors for Environment and Health” aims to develop a new generation of biosensors, functionalized, nano-structured and non, multiplexed, for the environment and for health.
In particular, there have been identified two broad areas of applicability of those devices:

• Monitoring of environmental pollutants that tend to bio-magnify within food chains, with consequent impact on human health;
• Molecular and clinical diagnostics, with particular regard to the evaluation of clinical markers, infectious agents, drugs for the treatment of chronic and degenerative diseases (i.e. Alzheimer).

In our society there is a growing demand for simple, rapid, efficient, reliable and analytical tools monitoring the environmental pollutants such as POPs (Persistent Organic Pollutants), of which the most representative are PAHs, PCBs, PBDEs, inorganic pollutants (i.e. heavy metals) and algal toxins.
Even in the field of molecular/clinic diagnostics there is an urgent need of having available analytical methods with similar characteristics to measure and evaluate clinical markers (i.e. tumor markers), the onset of immune-mediated diseases, infectious agents. In addition, ultrasensitive assays are required for the development of innovative non-invasive approaches for early prognosis of important diseases. The aim is the realization of a device that can also be used in ‘doctor office’ and / or in not hospital structures, becoming a “Point Of Care Testing ‘(POCT).
The development of electrochemical sensors and biosensors, with different types of transduction, in the areas of interest, representing the strength of the RUs proposing such a platform.

INBB researchers’ work has significantly contributed to the development of biosensors and bio-analytical methods (immunoassays and gene, luminescent cell biosensors, immunosensors, enzyme inhibition biosensors, DNA/RNA biosensors, methods for “high throughput screening” and immunohistochemistry and hybridization “in situ” methods for molecular imaging “in vivo”) with electrochemical, fluorescence, piezoelectric, plasmonic, and luminescent (bio, chemical, elettrochemical-, and thermochemical-luminescence) detection for applications in the biomedical, pharmaceutical , agribusiness, environmental and veterinary medicine. They also developed analytical devices suitable for portable applications in the field and/or in decentralized locations using portable equipment, often coupled with microfluidic systems.

INBB researchers’ commitment in the specific area is also reported from the activity reports submitted as part of the INBB National Conferences, among which are mentioned the most recent ones related to INBB IX National Conference, held in Rome on 21-22 October 2010 and X INBB National Conference, held in Rome on 22-23 October 2012, whose detailed programs and abstracts of the reports are on this website: www.inbb.it

The activities that the INBB researchers intend to implement in the near future are aimed at the development of two hybrid systems of (bio) sensors; the first one suitable for simultaneous detection of a large number of toxic or harmful substances (biological or chemical) present in the environment, the second one aimed at the identification of clinical and molecular targets of huge impact on human health.
Regarding the development of biosensors for the environment, each system will consist of a (bio) sensors platform based on different recognition of biological/chemical elements (aptamers, enzymes, antibodies, cells) and different transduction principles (chemical luminescence, fluorescence, plasmonics, electrochemical, piezoelectric). The systems will be designed according to this architecture will be very versatile and easily adaptable to a “multiplexed” analysis aimed at simultaneous detection of multiple targets in different matrices. In this way you can get information at right angles to each specific compound of interest, optimize the dynamic range and the sensitivity of the measurement in order to avoid possible effects of interfering matrix to be analysed.
The designed system for monitoring of environmental pollutants will operate on two levels. A first non-selective level, based on cell biosensors will be used as an early warning system and will have a high probability of detecting the substances of interest. The first level of monitoring is therefore based on the observation of the toxic effects of the analytes on various cell lines (bioluminescent wild-type or genetically modified), suitably coupled to optical systems. In particular, using different types of cells, in parallel, it will be possible to monitor a wide variety of toxic compounds (i.e. hepatotoxic or neurotoxic).
A second level, more discriminating, will take a platform with various biosensors and will aim to confirm the presence of the compounds of interest, allowing analysis of (semi) quantitative and with a low probability of false positives. This second level of monitoring, aimed at creating a selective detection of a “panel” of target molecules or classes of targets, will confirm the information obtained in the first level of monitoring by minimizing the likelihood of false positives. This stage is based on:

1. the direct determination of electro-active compounds (i.e. heavy metals) using functionalized electrode materials and/or nano-structured;
2. enzyme inhibition biosensors, immune-sensors/apta-sensors, and cell biosensors, using different types of transducer. Also in this case, the biological element (enzyme, antibody, aptamer, cells) that serves as a selector signal, may be coupled to nano-structured material in order to improve the “performance” of the analytical biosensors themselves.

This second level, if coupled with microfluidic systems, will allow continuous monitoring of the environmental concentrations, thus allowing the real-time assessment of the effectiveness of any decontamination procedures are in place.

Regarding the development of biosensors for molecular/clinic diagnostics, it will operate essentially on the second monitoring level with the objective of achieving a multi-parameters integrated platform that can also be used in ‘doctor office’ and/or in non-hospital facilities becoming a “Point Of Care Testing ‘(POCT). The possibility to implement ultrasensitive detection assays for the early detection of cancer biomarkers will be also investigated. In this way, it will be possible to carry out a screening of a larger population, make a timely early diagnosis of diseases (cancer, chronic and degenerative diseases, immune-mediated), predict therapeutic strategies suitable and customized, and monitor the effectiveness of the therapy itself.
The realization of the project will require a multidisciplinary approach: the specific skills of the participants will enable the development of (bio) sensors with different types of transduction, designed to be integrated in two hybrid sensing targeted platforms, a monitoring of environmental pollutants; the other for the early diagnosis of serious diseases. The advantage of these systems will revolutionize not only the environmental and diagnostic approach, but also the follow-up of a possible drug therapy or process of environmental remediation. In this way, it will be possible to optimize patient care or interventions of rectification simultaneously reducing environmental costs (in particular for all those who need treatments or reclamation of a continuous monitoring of certain parameters).

Several INBB RUs operating in the Platform on “Innovative biosensors for environment and human health” in the first place Bologna, Catania, Florence, II University of Naples, Padua, Rome Tor Vergata, Roma Tre Research Units.

SOME RECENT PUBLICATIONS

• Esteban Fernández de Ávila B, Watkins HM, Pingarrón JM, Plaxco KW, Palleschi G, Ricci F. Determinants of the Detection Limit and Specificity of Surface-Based Biosensors Anal Chem. 2013; 85 (14): 6593–6597.
• Ricci F, Adornetto G, Palleschi G. A review of experimental aspects of electrochemical immunosensors. Electrochim Acta. 2012;84:74-83.
• Arduini F, Giudone S, Amine A, Palleschi g, Moscone D. Acetylcholinesterase biosensor based on self-assembled monolayer-modified gold-screen printed electrodes for organophosphorus insecticide detection. Sensors and Actuators B 2013; 179: 201-208.
• Calvo Quintana J, Arduini F, Amine A, van Vezen K, Palleschi G, Moscone D. Part two: Analytical optimization of a procedure for lead detection in milk by means of bismuth-modified screen-printed electrodes. Anal Chim Acta 2012; 736: 92-99.
• Roda A, Cevenini L, Borg S, Michelini E, Calabretta MM, Schuler D: Bioengineered bioluminescent magnetotactic bacteria as a powerful tool for chip-based whole-cell biosensors. Lab Chip 2013, 13, 4881-4889.
• Cevenini L, Michelini E, D’Elia M, Guardigli M, Roda A: Dual-color bioluminescent bioreporter for forensic analysis: evidence of androgenic and anti-androgenic activity of illicit drugs. Anal Bioanal Chem 2013, 405, 1035-1045.
• Casolari S, Roda B, Mirasoli M, Zangheri M, Patrono D, Reschiglian P, Roda A: Gravitational field-flow fractionation integrated with chemiluminescence detection for a self-standing point-of-care compact device in bioanalysis. Analyst 2013, 138, 211-219.
• Mirasoli M, Buragina A, Dolci LS, Simoni P, Anfossi L, Giraudi G, Roda A: Chemiluminescence-based biosensor for fumonisins quantitative detection in maize samples. Biosens Bioelectron 2012,32, 283-287.
• Mariani S. & Minunni M. Surface plasmon resonance applications in clinical analysis, Anal Bioanal Chem DOI 10.1007/s00216-014-7647-5.
• Laschi S., Palchetti I., Marrazza G., Mascini M., Innovative Electrodes to Control Trace Metal Ionization Used to Treat Pathogens in Water Distribution Systems, 2012 LNEE, Vol 109, 25-30
• Ilkhani, H.; Mascini, M.; Marrazza, G., The potential affibodies in new cancer marker immunosensors. 2012; Vol. 109 LNEE, 15-18.
• Ermini, M.L., S. Mariani, S. Scarano, D. Campa, R. Barale, M. Minunni, Single Nucleotide Polymorphism detection by optical DNA-based sensing coupled to whole genomic amplification. Analytical Bioanalytical Chemistry, Wiley-VCH, Ed. A. Roda, 2013, 405, 985-993.
• BottiniG., Losito F., Arienzo A., Priolisi F.R., Mari A., Visca P. and Antonini G. .A new method for microbiological analysis that could be used for Point-Of-Care Testing (POCT). The Open Emergency Medicine Journal, 2013, 5, 13-15. DOI: 10.2174/1876542401305010013