ElectroSpray Immobilization of LAccase foR cANnabinoids deTEction - ESILARANTE (PRIN 2022)

The main idea at the base of ESILARANTE project is to characterize and develop a portable sensor platform for tetrahydrocannabinol (THC) detection based on laccase biosensors produced through the green, low-cost and environment-friendly ElectroSpray Deposition (ESD) technique. The commercialization of such a platform will push the use of biosensors for forensic analysis and on-road detection beyond the current state of the art. The development of selective, ecofriendly, low cost, stable and recycling biosensors in different environments such as atmosphere, food industry or in clinical analyses is strongly required. Among the most studied bio-components, the enzymes play a leading role, being a class of selective and sensitive biological recognition elements. Thus the fabrication of enzyme biosensors based on laccase has attracted a huge interest in the last years since this, robust and cheap enzyme, is used in several fields. In the fabrication process, enzyme immobilization strategy is a key factor to develop an efficient tool with appropriate performances. Recently the PI has manufactured new promising amperometric laccase-based biosensors with unprecedented reuse and storage capabilities using the ambient ESD methodology as immobilization technique. The biosensors have been electrochemically tested to prove that such an immobilization technique is suited to manufacture high performance devices. Thanks to an interdisciplinary effort the first goal is to achieve an understanding at the molecular level of these new promising biosensors, their stability and reuse by using different spectroscopies, microscopies and analytical techniques. This would allow to make a step forward in the development of new green and renewable systems to reduce pollution, which is at the core of some pillars of the Horizon Europe program and of strong relevance in the NextGenerationEU agenda. The second goal is to apply laccase-based biosensors for the development of a portable and cost-effective platform for the detection of cannabinoids in capillary blood samples. While cannabinoid sensing protocols are well established in lab techniques, miniaturized and portable sensors are still not well developed. Electrochemical devices promise to reduce greatly the analysis time per sample and to lower dramatically the costs while allowing sensitivity and miniaturization. To face the challenges set by these ambitious goals, a consortium has been set up consisting of two units, CNR and UNIBO. The manufacture of the biosensors together with the electrochemical and physico-chemical characterization will be done at the CNR. These activities will be complemented at UNIBO through established mass-spectrometric methodologies, for a complete analytical validation of the THC biosensor and to characterise the laccase solution used for the ESD, which will help us to disentangle the reasons behind such an optimal immobilization.