Sabato D’Auria Istituto di Biochimica delle Proteine Consiglio Nazionale delle Ricerche

1. Progettazione e Sviluppo di innovativi Nanosensori a Fluorescenza per la Sicurezza e la Difesa Nazionale Sabato D’Auria Istituto di Biochimica delle Proteine Consiglio Nazionale delle Ricerche s.dauria@ibp.cnr.it Ricerca e Tecnologia per la Sicurezza. Auditorum di Finmeccanica, Roma 10 Novembre, 2005

2. analytes Biological recognitionelement transducer amplifier microelectronics Biological Recognition Element Enzyme Protein Antibody Signal Transducer Optical Electro-Chemistry Calorimetric Roma 10 Novembre, 2005

3. Why Proteins, but not Chemical Probes for sensing analytes??? The molecular complexity of chemical sensors quickly increases for complex biochemical analytes. Even if an appropriate probe could be designed and synthesized, there is no guarantee of useful spectral changes, adequate water solubility, and right affinity constants. The sensor showed on the next slide illustrates the complexity of fluorescence chemical probes. Roma 10 Novembre, 2005

4. The Complexity of a Fluorescence Chemical Probe Structure of the chemical probe used for fluorescence detection of AMP Roma 10 Novembre, 2005

5. Thermostable proteins as long-term stability probes: example of glucose sensing by mesophilic and thermophilic glucokinase Roma 10 Novembre, 2005

6. Third millennium biosensors will have the following features: - Long-term Stability - Non-Consuming analyte - Implantable - Wireless Roma 10 Novembre, 2005

7. Nanotechnology approach for stable protein-based nanosensors Selective binding of proteins to irradiated PS regions with submicrometer resolution. (2005 International Patent Pending) Roma 10 Novembre, 2005

8. Advanced Robot Machine for High Density Nano-patterning Sensing Biomolecules on Chips Roma 10 Novembre, 2005

9. Thermostable Glucose-Binding Protein from P. horikoshii for glucose sensing Roma 10 Novembre, 2005

10. Multi-analyte assay by nano-patterning biomolecules specifically labeled Roma 10 Novembre, 2005

11. Fluorescence Platform for advanced sensing methodologies Roma 10 Novembre, 2005

12. Family doctor and/or Hospital Labeled proteins immobilized on the chip Wireless communication Coenzyme-depleted enzymes and proteins belonging to the “binding-protein” family can be used as specific probes for the realization of non-consuming analyte fluorescence biosensors for analyses of high social interest. Roma 10 Novembre, 2005

13. Wireless biosensors for homeland security Roma 10 Novembre, 2005

14. Chips at Work Fluorescence emission of a biomolecule immobilized on a porous silicon chip. Fluorescence variations of Acetylcholine immobilized on a Porous silicon chip upon binding ammonium nitrate Roma 10 Novembre, 2005

15. Conclusions The use of specific biomolecules as fluorescence probes results in the design of advanced highly stable and sensitive nanosensors for analyses of high social interest. Fundings • - Consiglio Nazionale delle Ricerche - Commesa: Diagnostica Avanzata ed Alimentazione • - Regione Campania Centro di Competenze in Biotecnologie Industriali • - NATO Grant PST CLG 981025 • - NATO Grant LST 978934 • - Ministero Affari Esteri - Italia- Corea del Sud – Grant N. R1 • - Ministero Affari Esteri – Italia - Austria- Grant N. 4-2004 • Grant CNR-Kosef Roma 10 Novembre, 2005

16. Collaborations - Selex Sistemi Integrati, Dr. Carlo Falessi (MIUR Project, Submitted 2005) - Harvard University, Prof. J. Collier (Progetto Italy-USA, MIUR, Submitted 2005) - University of Texas, USA, Dr. I. Grycznyski (Core collaboration) - Russian Academy of Science (FIRB Project, with Selex SI, Submitted 2005) - ISS, Inc, Illinois, USA, Dr. B. Barbieri (Core Collaboration) - University of Prague, Dr. P. Herman (NATO) - University of Pussan, Korea, Dr. J. Kang (CNR; Kosef) - Institute Galileo Ferraris, Torino, Dr. A. Rossi (Core Collaboration) - Institute of Microelectronics, Dr. L. DeStefano (Core Collaboration) - University of Graz, Austria, Dr. B. Nidetzky (International Project) - Neotron, Spa, Modena, Italy, Dr. M. Gatti (Collaboration Project) Roma 10 Novembre, 2005

17. The 2004-2005 Lab’s Publications on Design and Development of Advanced Sensing Methodologies - Odor-binding protein as probe for a refractive index-based biosensor: new perspectives in biohazard assessment. S. D’Auria, A. Varriale, M. Staiano, V. Scognamiglio, M. Rossi, S. Campopiano, V. Cennamo, L. Zeni , (2005) Proteins Peptide Letters (in press) - Advanced Protein-Based Biosensors: Glucose Biosensors as a Model for Analyses of High Social Interest M. Staiano, P. Bazzicalupo, M. Rossi, and S. D’Auria Molecular BioSystems (2005) DOI:10.1039 - Porous silicon-based optical microsensor for the detection of L-Glutamine L. De Stefano, L. Rotiroti, I. Rendina, L.Moretti, V. Scognamiglio, M. Rossi, and S. D’Auria (2005) Biosensors Bioelectronics (in press) - Writing 3D protein nanopatterns onto a silicon nanosponge S. Borini, S. D’Auria, M. Rossi, A. M. Rossi (2005) Lab-on-a-Chip Oct;5(10):1048-52 - Advanced Fluorescence Biosensors for Diabetic patients S. D’Auria, V. Scognamiglio, M. De Champdore’, M. Staiano, G. Ghirlanda, M. Rossi (2005) Topics in Fluorescence Spectroscopy,10, Plenum Press, New York, USA - A putative thermostable sugar-binding protein from the archaeon Pyrococcus horikoshii as a probe for the development of a fluorescence biosensor for diabetic patients. M. Staiano, MR Sapio, V. Scognamiglio, A. Marabotti, AM Facchiano, P. Bazzicalupo, M. Rossi, and S. D’Auria Biotechnology Progress(2004) 5:1572-80. - Protein-based biosensors for diabetic patients V. Scognamiglio, M. Staiano, M. Rossi, and S. D’Auria. Journal of Fluorescence (2004) 14,5,491-498. Roma 10 Novembre, 2005

18. Personnel Prof. Mose’ Rossi, Director Dr. Vincenzo Aurilia, Molecular Geneticist Dr. Maria Staiano, Biochemist Dr. Marcella de Champdore’, Chemist Dr. Viviana Scognamiglio, Biologist Dr. Antonio Varriale, Biotechnologist Dr. Antonietta Parracino, Chemist Mr. Gianluca Aquino, Student Mrs. Annalisa Vitale, Student Mrs. Immacolata Cocozza, Student Roma 10 Novembre, 2005