Sensors and Biosensors group

A.P.F. Turner

Aristotele University of Thessaloniki (Greece) Bucharest University (Romania) Cranfield University (UK) Brno University (Cezch Republic) Cluj-Napoca University (Romania)

Marco Mascini

•Post-docs

Francesca Bettazzi Simona Scarano

•PhD students

Andrea Ravalli Diego Voccia Stefano Mariani

•Visiting professors •Erasmus link

Giovanna Marrazza Ilaria Palchetti Maria Minunni

1. Development and evaluation of sensors and biosensors based on the

coupling of electrochemical, optical and piezoelectric sensors with enzymes, antibodies, bacteria, whole tissues and nucleic acids

2. Solving analytical problems in clinical chemistry, experimental medicine, food chemistry and environmental analysis

3. Development of new immobilization chemistries of biomolecules (enzymes, proteins, nucleic acids, etc.) and analytical procedures suitable for use with biosensor devices, i.e. flow injection analysis, flow systems, and microdialysis

Group’s expertise

Autolab Potentiostat/Galvanostat (Metrohm, NL))

Palm Sens electrochemical sensor interfaces (NL)

Home made screen-printed electrodes

Electrochemical instrumentations

Scanning electrochemical microscope (SECM)

Microfluidic system

Electrochemical & Piezoelectric instrumentations

GRAVI - CHIP Immuspeed

Universal sensor

Seiko quartz microbalance

Elbatech QC magic (4 channels)

• 9.5 MHz AT-Cut quartz crystal (14mm); Gold evaporated (42.6mm2 area) on both sides Crystal Manufactoring (USA)

Fully automated Biacore X™(GE Healthcare)

Spreeta ™ (Texas Instruments Inc. USA)

SPR-imaging (Genoptics- France)

Optical instrumentations

Recent applications and main research topics

Chair: J.P. Aimè, UMR 5248 CBMN CNRS -Universite Bordeaux- COST Action TD 1003 "BioInspired Nanotechnologies": http://www.bioinspired-nano.eu/en/

Italy MC members: M. Minunni, Università di Firenze,

Research: • Développement d’un biocapteur à base d’un nouvel aptamère pour la diagnostique dans la pathologie d’Alzheimer • Sviluppo di biosensori di affinità a base di un nuovo recettore aptamerico per la diagnostica molecolare della malattia di Alzheimer Parternership 'Universitè di Grenoble I, Joseph Fourier (Prof. Eric Peyrin). Investigator: M. MINUNNI

•Drug discovery: Studies of amyloid fibrills aggregation and its inhibition by drugs (Alzheimer treatment?) X. Cheng, V.i Wing S. Hung, S. Scarano, M. Mascini, M. Minunni,* and K- Kerman*, Label-free methods for probing the interaction of Clioquinol with Amyloid-β, Anal. Methods, 2012, 4, 2192-2192

• DNA-based sensing for target sequence or mismatch detection, i.e. Single nucleotide polymorphism (SNPs); Molecular diagnostic, theranostic, tailored therapy Ermini, M.L., S. Mariani, S. Scarano, M. Minunni, Biosensor and Bioelectronics, 40 (1), 2013, 193-199; Ermini, M.L., S. Mariani, S. Scarano, D. Campa, R. Barale, M. Minunni*, Analytical Bioanalytical Chemistry,, 2013, 405, 985-993

• Aptasensor: Aptamer –ligand interaction detection of MMP9; Molecular diagnostic •J.J Toulmé, E. Dausse, S. Scarano, F. Crispo , M.. Minunni, Aptamers for the detection of biomarkers., NanoBioEurope, 10-12 June 2013, Toulouse,

• Immunosensors: Ag-Ab interaction; Molecular diagnostic S. Scarano, A. Vestri, M. L. Ermini and M. Minunni, SPR detection of human Hepcidin-25: a critical approach by immuno- and biomimetic-based biosensing, Biosensor and Bioelectronics, 40 (1), 2013, 135-140;

• Improving analytical performances by nanostructuring S. Mariani, M.L. Ermini, S. Scarano, F. Bellissima, D. Berti, M. Bonini*, M. Minunni*, Nanotechnology Coupled to Biosensing: Looking for improved Analytical performances with application to DNA-Based Sensing, Microchimica Acta, 2013, 180, 1093-1099

Case studies:

Single nucleotide polymorphisms high sensitive

detection on human DNA – with surface plasmon

resonance Imaging

S. Mariani1, S. Scarano1, R. Barale2,

M. Minunni1

1Dipartimento di Chimica “Ugo Schiff”, Università degli Studi di Firenze (Italiy) 2 Dipartimento di Biologia, Università degli Studi di Pisa (Italiy)

8-9/10/2013 Thematic COST meeting: “Nano-scales arrangements of proteins,

aptamers and other nucleic acid structures - and their potential

applications"

Surface Plasmon Resonance (SPR)

Biochip

SPR imaging (SPRi)

Sensing Area

RU

Time

Sensorgram

Biochip Sensing Areas: Regions of Interest (ROIs)

Sensorgrams

Time

ΔR%

Array of Light Sensitive Diodes (LSD)

Charge Coupled Device (CCD)

SPR IMAGING (SPRI) FOR BIOSENSING: A LABEL-FREE, MULTIARRAY PLATFORM FOR THE DETECTION OF BIOAFFINITY INTERACTIONS

S. Scarano, M.Mascini, APF Turner and M.Minunni, Surface Plasmon Resonance Imaging for Affinity-Based Biosensors, Biosensors and Biolectronics, 2010, 25, 5, 957-966 G. Spoto, M. Minunni, Surface Plasmon Resonace imaging: what next? Perspective, J. Phys. Chem. Lett., pp 2682–2691, 2012

Biochips and Array Preparation

B

B

B

B

B*

B*

B*

H

H

H

HH*

H*

H*

blank blank

A B C

D

Biochip view by CCD before interaction

Biochip design

Probe 3

Probe 1

Probe 2

Negative controls

Bioreceptors Array

Glass prism

Gold layer

Resonance couplers: Glass prisms or slides coated with gold

Array of wells

PDMS layer

Microsyringe

Surface Plasmon Resonance imaging DNA sensor

Principle and instrument

• Interaction light\electron → Plasmon excitation → SPR

• DNA Probes immobilization

• Affinity interaction → DNA Probe \ DNA Analyte → Signal (Sensorgram + Digital Image)

Sensorgram

(Real time)

Digital Image

(Multi-array)

OUTPUTS

Topic and case study

•SNP discrimination on non-amplified hDNA

Case study

• Gene ABCB1→ ATP-binding cassette transporter

• SNP rs 1045642 → Protein expression ⤍ Opioids drug absorption

Pharmacogenomic interest

Patient’s SNP ⤍ Tailored drug therapy with opiods (Pain therapy)

Ermini, M.L., S. Mariani, S. Scarano, M. Minunni, Biosensor and Bioelectronics, 40 (1), 2013, 193-199; Ermini, M.L., S. Mariani, S. Scarano, D. Campa, R. Barale, M. Minunni*, Analytical Bioanalytical Chemistry, Wiley-VCH, Ed. A. Roda, 2013, 405, 985-993

Mondiali di ciclismo a Firenze……