Leader: Paolo Matteini (CNR-IFAC); Other collaborator(s): Aloisi, Capone, Forleo (CNR-IMM); Baldini, Berneschi, D'Andrea, Matteini (CNR-IFAC)
Specific studies aimed at advancing established and emerging analytical techniques toward the identification of characteristic biomarker patterns in diverse human matrices will be pursued. A first activity concerns the setup of platforms based on Raman and on fluorescence techniques for capturing the optical fingerprint and for multianalyte detection in real samples. Another activity focuses on volatilomics through the development of an electronic nose trained by GC/MS volatilome data and the optimization of a Solid Phase Microextraction-GC/MS method for analyzing volatile organic compounds profiles in liquid biosamples. As complementary activity a sweat collection tool based on the integration of microchannels, reservoirs, and nano-porous textile sponges will be developed.
Brief description of the activities and of the intermediate results
The activity involves the development of a surface-enhanced Raman scattering (SERS) sensing platform for optical fingerprinting of biological fluids, including serum. Nanostructured silver spotted, flexible and disposable substrates are under development to generate an optical enhancement in the Raman signal and facilitating the use of small biofluid aliquots according to a time-efficient procedure. Another activity centres on an optical platform utilizing long period gratings (LPGs) for biomarker sensing. The platform feature multianalyte detection using different LPGs on a single fiber, each with distinct resonance wavelengths and containing a unique biological recognition element selective to a specific biomarker. About the volatilomics activity, the SPYROX electronic nose device development is ongoing, utilizing chemoresistive gas sensors trained by volatilome data obtained through GC/MS. SPYROX serves as a digital fingerprinting tool for the volatilome of selected biofluids. As complementary acivity, the exploration of cutting-edge technologies and materials for skin-interfaced sweat collection systems. This involves the integration of microchannels, reservoirs, and a combination of nano-porous textile sponges to facilitate sweat collection during on-body wear. These activities can support intervention study of WP1 (In-Tempo) on a subpopulation and that of WP3 (I COUNT) on the total population for secondary explorative objectives.
Main policy, industrial and scientific implications
Promotion of interdisciplinary research: The proposed activity may call for funding policies that encourage collaboration across diverse fields (biochemistry, engineering, nanotechnology), fostering the development of emerging technologies like SERS devices and wearable sweat collection systems;
Growth in diagnostic device markets: The introduction of advanced platforms like that based on SERS and SPYROX could drive innovation in wearable and portable diagnostic tools, opening new market opportunities in healthcare technology.
Enhanced biomarker detection
The use of multi-analyte platforms (SERS, LPG) enables more precise and simultaneous detection of biomarkers, advancing research in early disease diagnosis and personalized medicine
The proposed platforms are at an advanced stage of development, undergoing validation tests or early-stage applications in clinical research settings.
SERS platform: The development of nanostructured silver substrates has been finalized, optimizing their performance for enhanced optical fingerprinting. The SERS platform has been tested on a wider range of biomolecules (HSA, BSA, Lysozime, Tranferrrin, Amyloid b42. etc.) to validate its effective and sensitive detection capabilities.
LPG sensor: The multianalyte detection capabilities of LPG sensors are in progress by developing the algorithm to extract the information derived by the signal modulation caused by the interaction of the detected molecules with the different sensing biolayers deposited on the LPGs. In parallel the microfluidic circuit associated with the flow cell containing the fibre with the inscribed LPGs on it has been further developed using suitable syringe pumps and 3-way valves. The developed platform will be tested on some of the biomolecules already tested on the SERS platform.
SPYROX electronic nose and GC/MS analysis: The development of an electronic nose devoted to human volatilome fingerprinting was completed. The device, code-named SPYROX, adopts an array of 8 metal-oxide (MOX) gas sensors and it can analyze response signals from different matrices (multi-matrix samples), dealing with exhaled breath and headspace analysis of human biological samples. The functionality of the device was proved by a classification test of chemical standards and VOC mixtures as well. The device is currently being used in the analysis of blood and urine jointly with volatilome analysis by SPME-GC/MS on a sample population within the framework of scientific collaborations.
Sweat collection tool: a patch-based multilayer system has been designed and medical-grade polymeric films with hydrophilic/hydrophobic properties are now under consideration to increase the volume of collected sweat in a leakage-free manner.
The proposed platforms underwent further optimization improving their use in real settings.
SERS platform: The SERS platform has been further optimized in the form of a wearable SERS-active chip by the use of biocompatible and transparent adhesive tapes adhered on each side of the substrate and proper modification with a small conduct allowing the direct flow of the biofluid (such as sweat) to the sensor.
LPG sensor: The activity dedicated to determining the algorithm to extract data from two LPGs written along the same fibre in order to allow the simultaneous measurement of two analytes is still ongoing; the flow-cell is ready to be used for the determination, at the moment, of a single analyte.
SPYROX electronic nose and GC/MS analysis: The electronic nose has been further configured for fingerprinting the human volatilome based on the headspace analysis of human biological samples (as urine, blood and exhaled breath collected in Tedlar gas sampling bag). The activity of using the device in the analysis of human biological samples, jointly with the analysis of volatiles by SPME-GC/MS, is still ongoing; urine samples have been made available within scientific collaborations external to Age.it, awaiting the biological samples that will be collected in the In-Tempo study protocol in WP1 and in the ICount study protocol in WP3 for secondary exploratory objectives.
Sweat collection tool: natural polysaccharide blends (including alginate and chitosan) have been tested in the patch integrated microfluidic channel and the characterization is still ongoing.
Scientific publications:
Conferences: