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

Sabato D’AuriaIstituto di Biochimica delle ProteineConsiglio Nazionale delle Ricerche

[email protected]

Progettazione e Sviluppo di innovativi Nanosensori a Fluorescenza per la Sicurezza e la Difesa Nazionale

Ricerca e Tecnologia per la Sicurezza. Auditorum di Finmeccanica, Roma 10 Novembre, 2005

Biological recognitionBiological recognition elementelement

transducertransducer

amplifieramplifier

microelectronicsmicroelectronics

analytesanalytes

Biological Recognition Biological Recognition ElementElement EnzymeProtein

Antibody

Signal TransducerSignal Transducer Optical

Electro-ChemistryCalorimetric

Roma 10 Novembre, 2005

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.

Why Proteins, but not Chemical Probes for sensing analytes???

The sensor showed on the next slide illustrates the complexity of fluorescence chemical probes.


O ON

NN

HN

O

Pr

Pr

PrNH

O

O

O

O

N

H

H

O

O

O

P O

O

N

NN

N

NHH

Pr

Pr

PrN

O

OH

O

NH

OH

Structure of the chemical probe used for fluorescence detection of AMP


The Complexity of a Fluorescence Chemical Probe


Thermostable proteins as long-term stability probes: example of glucose sensing by mesophilic and thermophilic glucokinase

Third millennium biosensors will have the following features:

- Long-term Stability

- Non-Consuming analyte

- Implantable

- Wireless


Selective binding of proteins to irradiated PS regions with submicrometer resolution. (2005 International Patent Pending)

Nanotechnology approach for stable protein-based nanosensors



Advanced Robot Machine for High Density Nano-patterning Sensing Biomolecules on Chips


Thermostable Glucose-Binding Protein from P. horikoshii for glucose sensing


Multi-analyte assay by nano-patterning biomolecules specifically labeled


Fluorescence Platform for advanced sensing methodologies

Family doctor and/orHospital

Wireless communicationCoenzyme-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.

Labeled proteins immobilized on the chip



Wireless biosensors for homeland security

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

Chips at Work


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



- 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)

Collaborations

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 InterestM. Staiano, P. Bazzicalupo, M. Rossi, and S. D’AuriaMolecular 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 nanospongeS. Borini, S. D’Auria, M. Rossi, A. M. Rossi(2005) Lab-on-a-Chip Oct;5(10):1048-52- Advanced Fluorescence Biosensors for Diabetic patientsS. 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 patientsV. Scognamiglio, M. Staiano, M. Rossi, and S. D’Auria.Journal of Fluorescence (2004) 14,5,491-498.


Personnel

Prof. Mose’ Rossi, DirectorDr. Vincenzo Aurilia, Molecular GeneticistDr. Maria Staiano, BiochemistDr. Marcella de Champdore’, ChemistDr. Viviana Scognamiglio, BiologistDr. Antonio Varriale, BiotechnologistDr. Antonietta Parracino, ChemistMr. Gianluca Aquino, StudentMrs. Annalisa Vitale, StudentMrs. Immacolata Cocozza, Student


Documents

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