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
[email protected]
Ricerca e Tecnologia per la Sicurezza. Auditorum di Finmeccanica, Roma 10 Novembre, 2005
Biological Recognition
Element
Signal Transducer
Optical
Electro-Chemistry
Calorimetric
Enzyme
Protein
Antibody
analytes
Biological recognition element
transducer
amplifier
microelectronics
Roma 10 Novembre, 2005
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
The Complexity of a Fluorescence Chemical Probe
O
Pr
H
NH
O
Pr
H
N
O
Pr
O
H
N
N
N
N
HN
N
O
O
Pr
Pr
Pr
NH
O
O
O
O
O P O
O
H
N
N
N
H
N
OH
O
O
Structure of the chemical probe used for fluorescence detection of AMP
Roma 10 Novembre, 2005
Thermostable proteins as long-term stability probes: example
of glucose sensing by mesophilic and thermophilic glucokinase
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Third millennium biosensors will have the
following features:
- Long-term Stability
- Non-Consuming analyte
- Implantable
- Wireless
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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
Advanced Robot Machine for High Density Nano-patterning
Sensing Biomolecules on Chips
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Thermostable Glucose-Binding Protein from P. horikoshii
for glucose sensing
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Multi-analyte assay by nano-patterning biomolecules specifically labeled
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Fluorescence Platform for advanced sensing methodologies
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Labeled proteins immobilized on the chip
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.
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Family doctor and/or
Hospital
Wireless biosensors for homeland security
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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
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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
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)
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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
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