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Biosensors and the Nanoscale Processing of Biomolecules Valtencir Zucolotto Laboratório de Nanomedicina e Nanotoxicologia Instituto de
Biosensors and the Nanoscale Processing of Biomolecules
Biosensors and the Nanoscale
Processing of Biomolecules
Valtencir Zucolotto Laboratório de Nanomedicina e Nanotoxicologia Instituto de Física de São Carlos - USP
Valtencir Zucolotto
Laboratório de Nanomedicina e Nanotoxicologia
Instituto de Física de São Carlos - USP

nanomedicina.com.br

BRASONN Nov/2011 zuco@ifsc.usp.br
BRASONN Nov/2011
zuco@ifsc.usp.br

Prof. Dr. Valtencir Zucolotto

 Biosensors  Basic Concepts in Biosensing  Classification  Immobilization Mechanisms  Detection Techniques 

Biosensors

Basic Concepts in Biosensing Classification Immobilization Mechanisms Detection Techniques

Applications of Biosensors Final Remarks

Analytical Parameters

 Biosensors  Basic Concepts in Biosensing  Classification  Immobilization Mechanisms  Detection Techniques 
 Biosensors  Basic Concepts in Biosensing  Classification  Immobilization Mechanisms  Detection Techniques 

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Biosensors

Biosensors  World Analytical Market is £ 12 bi/year  30% of that amount is related
  • World Analytical Market is £ 12 bi/year

  • 30% of that amount is related to “Health Care

Biosensors: 0.1% of HC market with na annual

increase rate of 60%

  • OBS: Around 6% of occidental people suffer from

diabetics and may benefit with the development of specific biosensors.

Source: http://www.lsbu.ac.uk/biology/enztech/biosensors.html

Biosensors  World Analytical Market is £ 12 bi/year  30% of that amount is related

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Biosensors CONTOUR® Blood Glucose Meter http://www.bayercarediabetes.com/us/prodserv/products/contour/index.asp nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br
Biosensors
Biosensors
Biosensors CONTOUR® Blood Glucose Meter http://www.bayercarediabetes.com/us/prodserv/products/contour/index.asp nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br
Biosensors CONTOUR® Blood Glucose Meter http://www.bayercarediabetes.com/us/prodserv/products/contour/index.asp nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br

CONTOUR® Blood Glucose Meter

http://www.bayercarediabetes.com/us/prodserv/products/contour/index.asp

Biosensors CONTOUR® Blood Glucose Meter http://www.bayercarediabetes.com/us/prodserv/products/contour/index.asp nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Components of a Biosensor

Components of a Biosensor Analyte Immobilized biological material Signal Processing Transductor Output Reference nanomedicina.com.br Prof. Dr.
Analyte
Analyte
Immobilized biological material Signal Processing Transductor
Immobilized
biological material
Signal
Processing
Transductor

Output

Components of a Biosensor Analyte Immobilized biological material Signal Processing Transductor Output Reference nanomedicina.com.br Prof. Dr.

Reference

Components of a Biosensor Analyte Immobilized biological material Signal Processing Transductor Output Reference nanomedicina.com.br Prof. Dr.

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Classification of Biosensors
Classification of Biosensors
Classification of Biosensors Recognition Mechanism Antigen Bioreceptor Antibodies DNA Enzymes Interaction Signal Piezo/gravimetric Electrochemical Impedantiometric Transducer

Recognition Mechanism

Antigen Bioreceptor Antibodies DNA
Antigen
Bioreceptor
Antibodies
DNA

Enzymes

Interaction

Classification of Biosensors Recognition Mechanism Antigen Bioreceptor Antibodies DNA Enzymes Interaction Signal Piezo/gravimetric Electrochemical Impedantiometric Transducer

Signal

Piezo/gravimetric

Electrochemical

Impedantiometric

Transducer
Transducer

Optical (Luminescence, absorption, SPR)

Classification of Biosensors Recognition Mechanism Antigen Bioreceptor Antibodies DNA Enzymes Interaction Signal Piezo/gravimetric Electrochemical Impedantiometric Transducer

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Immobilization of the biological species on the transducer Y Y Y Y Y Y Y Y
Immobilization of the biological species on the transducer
Immobilization of the biological species on the
transducer
Y Y Y Y Y Y Y Y Transducer
Y
Y
Y
Y
Y Y
Y
Y
Transducer
Immobilization of the biological species on the transducer Y Y Y Y Y Y Y Y

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Biomolecules Immobilization

Biomolecules Immobilization . Entrapment . Covalent Binding . Cross-Linking . Electrostatic Interactions, H bonds . Specific

. Entrapment . Covalent Binding . Cross-Linking . Electrostatic Interactions, H bonds . Specific Inteactions: Avidin-Biotin . Incorporation in Lipidic Membranes

Biomolecules Immobilization . Entrapment . Covalent Binding . Cross-Linking . Electrostatic Interactions, H bonds . Specific

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Biomolecules Immobilization

Biomolecules Immobilization Immobilization Mechanism Biomolecule Packing Orientation Efficiency Accessibility Mobility Biological Activity nanomedicina.com.br Prof. Dr. Valtencir

Immobilization

Mechanism

Biomolecule Packing Orientation Efficiency Accessibility Mobility
Biomolecule
Packing
Orientation
Efficiency
Accessibility
Mobility

Biological

Activity

Biomolecules Immobilization Immobilization Mechanism Biomolecule Packing Orientation Efficiency Accessibility Mobility Biological Activity nanomedicina.com.br Prof. Dr. Valtencir

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Protein Immobilization via Layered Thin Films nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br
Protein Immobilization via Layered Thin Films
Protein Immobilization via Layered Thin
Films
Protein Immobilization via Layered Thin Films nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Immobilization via Thin Films

Immobilization via Thin Films The Electrostatic Layer-by-Layer Technique The Langmuir-Blodgett Technique Self Assembled Monolayers (SAMs) nanomedicina.com.br

The Electrostatic Layer-by-Layer Technique

The Langmuir-Blodgett Technique

Self Assembled Monolayers (SAMs)

Immobilization via Thin Films The Electrostatic Layer-by-Layer Technique The Langmuir-Blodgett Technique Self Assembled Monolayers (SAMs) nanomedicina.com.br

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Immobilization via Thin Films

Immobilization via Thin Films 1. The Langmuir Blodgett Technique -Fabrication of ultrathin (organized) films; -Amphiphilic spread
1. The Langmuir Blodgett Technique
1. The Langmuir Blodgett Technique

-Fabrication of ultrathin (organized) films;

-Amphiphilic

spread

on

compression

molecules

in

a

volatile

a

water

sub-phase

with

by

mobile

barriers

spontaneous process);

solvent

further

(not

a

Immobilization via Thin Films 1. The Langmuir Blodgett Technique -Fabrication of ultrathin (organized) films; -Amphiphilic spread

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Immobilization via Thin Films

Immobilization via Thin Films -After compression, the molecules can undergo phase transition forming a dense monolayer

-After compression, the molecules can undergo phase transition

forming a dense monolayer (Langmuir film);

-Requires sophisticated, costly equipment (Langmuir Trough).

Mobile Barriers Area per Molecule Surface Pressure
Mobile Barriers
Area per Molecule
Surface Pressure

KSV Instruments Application Note # 107

Immobilization via Thin Films -After compression, the molecules can undergo phase transition forming a dense monolayer

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Immobilization via Thin Films

Immobilization via Thin Films Monolayers may be transferred to a solid platform upon immersion and withdrawal

Monolayers may be transferred to a solid platform upon immersion and withdrawal of a solid substrate (Langmuir-Blodgett LB film).

Immobilization via Thin Films Monolayers may be transferred to a solid platform upon immersion and withdrawal

The films can be molecularly thin as in the case of phospholipds films. Highly control over film thickness.

KSV Instruments Application Note # 107

Immobilization via Thin Films Monolayers may be transferred to a solid platform upon immersion and withdrawal

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Immobilization via Thin Films

Immobilization via Thin Films Useful for incorporation of proteins in the lipidic films. Substrate nanomedicina.com.br Prof.

Useful for incorporation of proteins in the lipidic films.

Substrate
Substrate
Immobilization via Thin Films Useful for incorporation of proteins in the lipidic films. Substrate nanomedicina.com.br Prof.

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Immobilization via Thin Films

Immobilization via Thin Films 2. Self-Assembled Monolayers Monolayers spontaneously adsorbed on a solid surface Silanization: Adsorption
2. Self-Assembled Monolayers
2. Self-Assembled Monolayers
Monolayers spontaneously adsorbed on a solid surface
Monolayers spontaneously adsorbed on
a solid surface
  • Silanization: Adsorption of silanes on silica-based substrates

  • Thiolation: Adsorption of thiol-based molecules on specific (metallic) surfaces (ex: gold)

Immobilization via Thin Films 2. Self-Assembled Monolayers Monolayers spontaneously adsorbed on a solid surface Silanization: Adsorption

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Immobilization via Thin Films

Advantages

Immobilization via Thin Films Advantages Experimental simplicity and organization at the molecular level; Higly stable and
  • Experimental simplicity and organization at the molecular level;

  • Higly stable and dense monolayers;

  • Appropriate environment for biological applications: cell growth and tissue engineering;

  • Versatility: Various functionalized molecules can be used to functionalize the surface.

Immobilization via Thin Films Advantages Experimental simplicity and organization at the molecular level; Higly stable and

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Immobilization via Thin Films

Immobilization via Thin Films 2.1 Silanization Substrates: Glass, Quartz, Silicon. Self-organization is driven by polysilanes formation,

2.1 Silanization

Immobilization via Thin Films 2.1 Silanization Substrates: Glass, Quartz, Silicon. Self-organization is driven by polysilanes formation,

Substrates:

Glass, Quartz, Silicon.

Self-organization is driven by polysilanes formation, which

are linked to silanol groups ay

the surface

Immobilization via Thin Films 2.1 Silanization Substrates: Glass, Quartz, Silicon. Self-organization is driven by polysilanes formation,

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Immobilization via Thin Films

Immobilization via Thin Films 2.2 Functionalized Thiols Cysteine 3-Mercaptopropionic acid Substrates: Au, Pt, Cu, Ag. nanomedicina.com.br

2.2 Functionalized Thiols

Cysteine
Cysteine
3-Mercaptopropionic acid
3-Mercaptopropionic acid

3-Mercaptopropionic acid

3-Mercaptopropionic acid

Substrates:

Au, Pt, Cu, Ag.

Immobilization via Thin Films 2.2 Functionalized Thiols Cysteine 3-Mercaptopropionic acid Substrates: Au, Pt, Cu, Ag. nanomedicina.com.br

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Immobilization via Thin Films

Immobilization via Thin Films Applications in Biosensing: SAMs as linkers for Biomolecules Immobilization (protein, enzyme, DNA,

Applications in Biosensing: SAMs as linkers for Biomolecules Immobilization (protein, enzyme, DNA, antibody, peptide, etc.)

Y Y Y Y Y Transducer
Y
Y
Y
Y
Y
Transducer
Immobilization via Thin Films Applications in Biosensing: SAMs as linkers for Biomolecules Immobilization (protein, enzyme, DNA,

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Immobilization via Thin Films

Immobilization via Thin Films 2. Electrostatic Self-Assembly  Iler (1966): Assembly of Colloidal particles bearing opposite
2. Electrostatic Self-Assembly
2. Electrostatic Self-Assembly

Iler (1966): Assembly of Colloidal particles bearing opposite charges

Sagiv (1983): Chemically Adsorbed MultiLayers:

Required Bifunctionalization

Immobilization via Thin Films 2. Electrostatic Self-Assembly  Iler (1966): Assembly of Colloidal particles bearing opposite

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Immobilization via Thin Films

Immobilization via Thin Films Monolayer O Si Si Adsorption O O Solid Substrate Activated Monolayer Bilayer….

Monolayer

O
O

Si

Si

Adsorption

O O
O
O

Solid Substrate

Activated Monolayer

Bilayer…. O Si Si
Bilayer….
O
Si
Si

Ativaction

Immobilization via Thin Films Monolayer O Si Si Adsorption O O Solid Substrate Activated Monolayer Bilayer….
OH OH O Si Si
OH
OH
O
Si
Si
O O
O
O

Adsorption

Immobilization via Thin Films Monolayer O Si Si Adsorption O O Solid Substrate Activated Monolayer Bilayer….
O O O Si Si
O
O
O
Si
Si
O O
O
O
Immobilization via Thin Films Monolayer O Si Si Adsorption O O Solid Substrate Activated Monolayer Bilayer….

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Immobilization via Thin Films

Immobilization via Thin Films  Decher ´ s Approach (1992): Polymeric Layers Assembled via electrostatic Attraction
Immobilization via Thin Films  Decher ´ s Approach (1992): Polymeric Layers Assembled via electrostatic Attraction

Decher´s Approach (1992): Polymeric Layers Assembled via electrostatic Attraction

“Build up of organized multilayer

thin films based upon the

electrostatic interaction btween

polycationic and polyanionic

species”

Immobilization via Thin Films  Decher ´ s Approach (1992): Polymeric Layers Assembled via electrostatic Attraction

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

nanomedicina.com.br

Immobilization via Thin Films

Immobilization via Thin Films Simplicity and versatility nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br
Simplicity and versatility
Simplicity and versatility
Immobilization via Thin Films Simplicity and versatility nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br
Immobilization via Thin Films Simplicity and versatility nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Immobilization via Thin Films

Polyelectrolytes

Immobilization via Thin Films Polyelectrolytes PAH H ( C CH 2 ) n CH 2 NH

PAH

H

( C CH 2 ) n CH 2 NH 3 Cl
(
C
CH 2
) n
CH 2
NH 3
Cl
( ) CH PSS
(
)
CH
PSS

SO 3 - Na +

PDAC

[ ] - N + Cl H 3 C CH 3
[
]
-
N +
Cl
H 3 C
CH 3
H H C C n H SO 3 - Na +
H
H
C
C
n
H
SO 3 - Na +

PVS

PEI

H 2 H 2 H N C C n
H 2
H 2
H N
C
C
n
Immobilization via Thin Films Polyelectrolytes PAH H ( C CH 2 ) n CH 2 NH

Dendrimers

Immobilization via Thin Films Polyelectrolytes PAH H ( C CH 2 ) n CH 2 NH

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Immobilization via Thin Films

Immobilization via Thin Films Experimental Parameters that Influence Film Properties -Solution Concentration; -Ionic Strength; -Immersion time;

Experimental Parameters that Influence

Film Properties

-Solution Concentration;

-Ionic Strength;

-Immersion time;

-Drying

-Solution pH.

Immobilization via Thin Films Experimental Parameters that Influence Film Properties -Solution Concentration; -Ionic Strength; -Immersion time;

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Immobilization via Thin Films

Immobilization via Thin Films Applications in Biosensing nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br

Applications in Biosensing

Immobilization via Thin Films Applications in Biosensing nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br
Immobilization via Thin Films Applications in Biosensing nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Immobilization via Thin Films

Immobilization via Thin Films Adsorption Mechanisms 1) Films from highly charged polyelectrolytes: Adsorption via electrostatic interactios;
Adsorption Mechanisms
Adsorption Mechanisms

1) Films from highly charged polyelectrolytes:

Adsorption via electrostatic interactios; Molecules in an extended configuration: Very thin layers;
Adsorption via electrostatic interactios; Molecules in
an extended configuration: Very thin layers;

2) Films from partially charged polyelectrolytes:

Amount of charges controlled by pH; Highly control over layer thickness (pH-dependent)
Amount of charges controlled by pH; Highly control
over layer thickness (pH-dependent)
Immobilization via Thin Films Adsorption Mechanisms 1) Films from highly charged polyelectrolytes: Adsorption via electrostatic interactios;

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Immobilization via Thin Films

Immobilization via Thin Films 3) Films assembled via secondary interactions: Adsorption mainly driven by H-bond, Van

3) Films assembled via secondary interactions:

Adsorption mainly driven by H-bond, Van der waals,
Adsorption mainly driven by H-bond, Van der waals,
etc
etc

4) Films assembled via specific interactions:

Biotin/Avidin functionalized Antigens/antibodies systems, etc.
Biotin/Avidin
functionalized
Antigens/antibodies systems, etc.
molecules,
molecules,
Immobilization via Thin Films 3) Films assembled via secondary interactions: Adsorption mainly driven by H-bond, Van

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Transduction Methods nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br

Transduction Methods

Transduction Methods nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Analyte Immobilized biological material Signal Processing Transductor Output Reference nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br
Analyte
Analyte
Immobilized biological material Signal Processing Transductor
Immobilized
biological material
Signal
Processing
Transductor

Output

Analyte Immobilized biological material Signal Processing Transductor Output Reference nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br

Reference

Analyte Immobilized biological material Signal Processing Transductor Output Reference nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Transduction Methods

Transduction Methods Quartz Crystal Microbalance (QCM)  Monitoring the amount of adsorbed material in “real time”
Quartz Crystal Microbalance (QCM)
Quartz Crystal Microbalance (QCM)

Monitoring the amount of adsorbed material in “real time” upon changes in the oscilation frequency of a quartz crystal

  • Correlation between adsorbed mass and frequency given by Sauerbry

equation:

Eletrodo (Au) Eletrodo (Au) Anticorpo Anticorpo Proteína Proteína Substrato de cristal Piezelétrico Substrato de cristal Piezelétrico
 

Eletrodo (Au)

Eletrodo (Au)

Anticorpo

Anticorpo

Proteína

Proteína

 
Substrato de cristal Piezelétrico Substrato de cristal Piezelétrico Eletrodo (Au) Eletrodo (Au)

Substrato de cristal Piezelétrico

Substrato de cristal Piezelétrico

 
Substrato de cristal Piezelétrico Substrato de cristal Piezelétrico Eletrodo (Au) Eletrodo (Au)
 

Eletrodo (Au)

Eletrodo (Au)

 
q A   2 f 2 0 q  m     
q
A
 
2 f
2
0
q
 m
  
f

filme

q = Módulo de cisalhamento do quartzo, f 0 = Freq. natural oscilação quartzo, q = Densidade do cristal

Transduction Methods Quartz Crystal Microbalance (QCM)  Monitoring the amount of adsorbed material in “real time”

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Transduction Methods

Transduction Methods <a href=E nzyme: Alcohol dehydrogenase (ADH) from yeast (E.C. 1.1.1.1), molecular weight of 141-151 kDa m (ng) 1500 1250 1000 IDE/(PAMAM/ADH)n Side View 750 500 250 0 2 4 6 8 10 Deposition Number of bilayers 139.5 ng per bilayer 52.1 ng attributed to ADH. A.C. Perinotto et al. , Thin Solid Films, 2008 nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br " id="pdf-obj-32-4" src="pdf-obj-32-4.jpg">

Enzyme: Alcohol dehydrogenase (ADH) from yeast (E.C. 1.1.1.1), molecular weight of 141-151 kDa

m (ng) 1500 1250 1000 IDE/(PAMAM/ADH)n Side View 750 500 250 0 2 4 6 8
m (ng)
1500
1250
1000
IDE/(PAMAM/ADH)n
Side View
750
500
250
0
2
4
6
8
10
Deposition
Number of bilayers
139.5 ng per bilayer
52.1 ng attributed to ADH.

A.C. Perinotto et al., Thin Solid Films, 2008

Transduction Methods <a href=E nzyme: Alcohol dehydrogenase (ADH) from yeast (E.C. 1.1.1.1), molecular weight of 141-151 kDa m (ng) 1500 1250 1000 IDE/(PAMAM/ADH)n Side View 750 500 250 0 2 4 6 8 10 Deposition Number of bilayers 139.5 ng per bilayer 52.1 ng attributed to ADH. A.C. Perinotto et al. , Thin Solid Films, 2008 nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br " id="pdf-obj-32-19" src="pdf-obj-32-19.jpg">

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Transduction Methods

Transduction Methods Optical Detection  Colorimetry;  Photoluminescence (PL);  SPRs nanomedicina.com.br Prof. Dr. Valtencir Zucolotto
Optical Detection
Optical Detection
  • Colorimetry;

  • Photoluminescence (PL);

  • SPRs

Transduction Methods Optical Detection  Colorimetry;  Photoluminescence (PL);  SPRs nanomedicina.com.br Prof. Dr. Valtencir Zucolotto

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Transduction Methods

Transduction Methods Surface Plamon Resonance (SPR)  Based on the total internal reflection of ligth and
Surface Plamon Resonance (SPR)
Surface Plamon Resonance (SPR)
  • Based on the total internal reflection of ligth and resonance of the plasmon

band;

Variations in the local refractive index at the surface of a metallic thin film deposited on a dielectric;

Transduction Methods Surface Plamon Resonance (SPR)  Based on the total internal reflection of ligth and

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

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Transduction Methods

Transduction Methods Determination of Ka and Kd; Determination of the adsorbed mass; n  n 2

Determination of Ka and Kd;

Determination of the adsorbed mass;

n  n 2 f buffer ( mg m / )  d f dn dc
n  n
2
f
buffer
(
mg m
/
) 
d
f
dn dc

n f = film refracive index, d f = film thickness

dn/dc

~ 0.2 cm 3 /g; n f ~ 1,57

Transduction Methods Determination of Ka and Kd; Determination of the adsorbed mass; n  n 2

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Prof. Dr. Valtencir Zucolotto

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Transduction Methods

Electrochemical Methods
Electrochemical Methods
Transduction Methods Electrochemical Methods Monitoring of the redox reactions occurring between immobilized species and the analyte

Monitoring of the redox reactions occurring between immobilized species and the analyte

Amp Volt Cottrell Equation: Contra-Eletrodo 1 nFAD C 2 Eletrodo  i ( t ) 
Amp
Volt
Cottrell Equation:
Contra-Eletrodo
1
nFAD C
2
Eletrodo
i
( t ) 
Referência
d
1
(
t
)
2
Eletrodo de
Trabalho

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

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Transduction Methods

Transduction Methods 30 0.1 0.2 0.5 0.0 0.4 0.3 I / nA 10 15 20 25
30 0.1 0.2 0.5 0.0 0.4 0.3 I / nA 10 15 20 25 2400 35
30
0.1
0.2
0.5
0.0
0.4
0.3
I / nA
10
15
20
25
2400
35
0
5
1,2 nA
1,2 nA
50 s
-150.0n
-120.0n
I / A
Intervalo linear
t / s

Concentração de glicose / mmol L -1

Crespilho, et al., Electrochemistry Communications, 2006

Transduction Methods 30 0.1 0.2 0.5 0.0 0.4 0.3 I / nA 10 15 20 25

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Transduction Methods

Transduction Methods Field Effect Transistors (FETs) Platforms  Detection of analytes using the Field Effect Transistor
Field Effect Transistors (FETs) Platforms
Field Effect Transistors (FETs) Platforms

Detection of analytes using the Field Effect Transistor (FETs) Architecture

 Detection of analytes using the Field Effect Transistor (FETs) Architecture  Conversion: Biological Response (Conductance

Conversion: Biological Response (Conductance ) Variation

Electrical properties

Transduction Methods Field Effect Transistors (FETs) Platforms  Detection of analytes using the Field Effect Transistor
Transduction Methods Field Effect Transistors (FETs) Platforms  Detection of analytes using the Field Effect Transistor

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Transduction Methods

Impedanciometric Detection
Impedanciometric Detection
Transduction Methods Impedanciometric Detection  Variation in the dielectrical properties of a matrix containing the immobilized

Variation in the dielectrical properties of a matrix containing the immobilized biological molecules deposited between capacitor plates.

~ Eletrodos Metálicos Material Biológico
~
~
~ Eletrodos Metálicos Material Biológico

Eletrodos Metálicos

Material Biológico

~ Eletrodos Metálicos Material Biológico
~ Eletrodos Metálicos Material Biológico
~ Eletrodos Metálicos Material Biológico
Transduction Methods Impedanciometric Detection  Variation in the dielectrical properties of a matrix containing the immobilized

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Transduction Methods

Transduction Methods Impedanciometric Detection  Combination of capacitors: Interdigitated electrodes nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br
Impedanciometric Detection
Impedanciometric Detection
  • Combination of capacitors: Interdigitated electrodes

Transduction Methods Impedanciometric Detection  Combination of capacitors: Interdigitated electrodes nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br
Transduction Methods Impedanciometric Detection  Combination of capacitors: Interdigitated electrodes nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br
Transduction Methods Impedanciometric Detection  Combination of capacitors: Interdigitated electrodes nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br

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Prof. Dr. Valtencir Zucolotto

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Transduction Methods

Transduction Methods Capacitance Frequency Antígeno Anticorpo nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br
Transduction Methods Capacitance Frequency Antígeno Anticorpo nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br
Capacitance
Capacitance

Frequency

Antígeno Anticorpo
Antígeno
Anticorpo
Transduction Methods Capacitance Frequency Antígeno Anticorpo nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br

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Prof. Dr. Valtencir Zucolotto

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Application of biomolecules- containing films in biosensing nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br
Application of biomolecules- containing films in biosensing
Application of biomolecules-
containing films in biosensing
Application of biomolecules- containing films in biosensing nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br

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AuNPs / Enzyme complexes
AuNPs / Enzyme complexes

Detection of Glucose

AuNPs / Enzyme complexes Detection of Glucose ITO-(PVS/PAMAM-Au@CoHCF@GOx) nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br
AuNPs / Enzyme complexes Detection of Glucose ITO-(PVS/PAMAM-Au@CoHCF@GOx) nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br

ITO-(PVS/PAMAM-Au@CoHCF@GOx) 3

AuNPs / Enzyme complexes Detection of Glucose ITO-(PVS/PAMAM-Au@CoHCF@GOx) nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br

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AuNPs / Enzyme complexes

AuNPs / Enzyme complexes Amp Volt Contra-Eletrodo Eletrodo de Trabalho Eletrodo Referência Linear Response: 10-100 µmol
Amp Volt Contra-Eletrodo Eletrodo de Trabalho
Amp
Volt
Contra-Eletrodo
Eletrodo de
Trabalho

Eletrodo

Referência

AuNPs / Enzyme complexes Amp Volt Contra-Eletrodo Eletrodo de Trabalho Eletrodo Referência Linear Response: 10-100 µmol

Linear Response: 10-100 µmol L-1 Sensibility: 115,11 nA mmol L-1 LOD = 5,5 µmol L-1

Glucose Detection
Glucose Detection
30 0.1 0.2 0.5 0.0 0.4 0.3 I / nA 10 15 20 25 -120.0n 35
30
0.1
0.2
0.5
0.0
0.4
0.3
I / nA
10
15
20
25
-120.0n
35
0
5
I / A
Intervalo linear
t / s
2400
50 s
-150.0n
1,2 nA
1,2 nA

Concentração de glicose / mmol L -1

Crespilho, et al., Electrochemistry Communications, 2006

AuNPs / Enzyme complexes Amp Volt Contra-Eletrodo Eletrodo de Trabalho Eletrodo Referência Linear Response: 10-100 µmol

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Nanowires/Enzyme Complexes
Nanowires/Enzyme Complexes
Nanowires/Enzyme Complexes Contato elétrico 2 Eletrodo 3 alumina Deposição de ouro 1 Eletrodeposição nanofios 5 4
Contato elétrico 2
Contato
elétrico
2
Eletrodo 3
Eletrodo
3
Eletrodo 3

alumina

Deposição de ouro 1
Deposição de ouro
1
Contato elétrico 2 Eletrodo 3 alumina Deposição de ouro 1 Eletrodeposição nanofios 5 4 Remoção Alumina

Eletrodeposição nanofios

5

4 Remoção Alumina
4
Remoção
Alumina
orientados Nanofios 6
orientados
Nanofios
6
Nanowires/Enzyme Complexes Contato elétrico 2 Eletrodo 3 alumina Deposição de ouro 1 Eletrodeposição nanofios 5 4
Nanowires/Enzyme Complexes Contato elétrico 2 Eletrodo 3 alumina Deposição de ouro 1 Eletrodeposição nanofios 5 4

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Nanowires/Enzyme Complexes
Nanowires/Enzyme Complexes
Nanowires/Enzyme Complexes Glucose Detection Detection limit: 22 μM (at 0.0 V). Apparent Michaelis- Menten constant :

Glucose Detection

Nanowires/Enzyme Complexes Glucose Detection Detection limit: 22 μM (at 0.0 V). Apparent Michaelis- Menten constant :

Detection limit: 22 μM (at 0.0 V).

Apparent Michaelis- Menten constant : 2 mM

Crespilho, et al., J. Phys. Chem., 2009

Nanowires/Enzyme Complexes Glucose Detection Detection limit: 22 μM (at 0.0 V). Apparent Michaelis- Menten constant :

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AgNPs / Enzyme complexes
AgNPs / Enzyme complexes
AgNPs / Enzyme complexes Urea Detection PVA-AgNp Urease ITO PAni Urea injection (mmol L -1 )

Urea Detection

PVA-AgNp

Urease
Urease
AgNPs / Enzyme complexes Urea Detection PVA-AgNp Urease ITO PAni Urea injection (mmol L -1 )
AgNPs / Enzyme complexes Urea Detection PVA-AgNp Urease ITO PAni Urea injection (mmol L -1 )
ITO PAni
ITO
PAni
Urea injection (mmol L -1 ) 2.0 1.0 5.0 1.0 1.5 + NH 4 2 A
Urea injection (mmol L -1 )
2.0
1.0
5.0
1.0
1.5
+
NH 4
2 A
+
NH 4
+
+
PAni + + NH 4
diffusion
NH 4
e -
+
NH 4
+
0
50
100
NH 4
Time / s
+
PAni•NH 4

ITO/PAni/Urease

AgNPs / Enzyme complexes Urea Detection PVA-AgNp Urease ITO PAni Urea injection (mmol L -1 )

ITO/PAni/PVA-Ag/Urease

AgNPs / Enzyme complexes Urea Detection PVA-AgNp Urease ITO PAni Urea injection (mmol L -1 )
AgNPs / Enzyme complexes Urea Detection PVA-AgNp Urease ITO PAni Urea injection (mmol L -1 )

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Phthalocyanine/Enzyme Complexes
Phthalocyanine/Enzyme Complexes
Phthalocyanine/Enzyme Complexes Tyrosinase Mixed Langmuir Film ITO ITO Electrode covered with LB Immobilized Tyrosinase J. Mater
Tyrosinase
Tyrosinase
Mixed Langmuir Film

Mixed Langmuir Film

Tyrosinase Mixed Langmuir Film ITO ITO Electrode covered with LB Immobilized Tyrosinase

LB FIlm

Tyrosinase Mixed Langmuir Film ITO ITO Electrode covered with LB Immobilized Tyrosinase

ITO

Tyrosinase Mixed Langmuir Film ITO ITO Electrode covered with LB Immobilized Tyrosinase

ITO Electrode covered with LB Immobilized Tyrosinase

J. Mater Chem., 2011, 21, 4995

Phthalocyanine/Enzyme Complexes Tyrosinase Mixed Langmuir Film ITO ITO Electrode covered with LB Immobilized Tyrosinase J. Mater

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Phthalocyanine/Enzyme Complexes
Phthalocyanine/Enzyme Complexes
Phthalocyanine/Enzyme Complexes J. Mater Chem., 2011, 21, 4995 nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br
Phthalocyanine/Enzyme Complexes J. Mater Chem., 2011, 21, 4995 nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br

J. Mater Chem., 2011, 21, 4995

Phthalocyanine/Enzyme Complexes J. Mater Chem., 2011, 21, 4995 nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

 

FET Devices

FET Devices

Electrochemical Ion Sensitive FETs containig CNTs

 

and Penicilinase for Penicilin Detection

FET Devices  Electrochemical Ion Sensitive FETs containig CNTs and Penicilinase for Penicilin Detection Siqueira Jr.

Siqueira Jr. et al, Biosens. Bioelectron, 2009 Fernandes, EG et al, J. Phys. Chem C, 2010

PAMAM SWNT-COOH Penicillinase dendrimer
PAMAM
SWNT-COOH
Penicillinase
dendrimer
FET Devices  Electrochemical Ion Sensitive FETs containig CNTs and Penicilinase for Penicilin Detection Siqueira Jr.

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Prof. Dr. Valtencir Zucolotto

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FET Devices Ta O SiO p-Si Al EIS Architecture 6-bilayer PAMAM/SWNT LbL film Siqueira Jr. et

FET Devices

FET Devices Ta O SiO p-Si Al EIS Architecture 6-bilayer PAMAM/SWNT LbL film Siqueira Jr. et
FET Devices Ta O SiO p-Si Al EIS Architecture 6-bilayer PAMAM/SWNT LbL film Siqueira Jr. et
FET Devices Ta O SiO p-Si Al EIS Architecture 6-bilayer PAMAM/SWNT LbL film Siqueira Jr. et

Ta 2 O 5

SiO 2

p-Si

Al

EIS Architecture

FET Devices Ta O SiO p-Si Al EIS Architecture 6-bilayer PAMAM/SWNT LbL film Siqueira Jr. et

6-bilayer PAMAM/SWNT LbL film

Siqueira Jr. et al, Biosens. Bioelectron, 2009

FET Devices Ta O SiO p-Si Al EIS Architecture 6-bilayer PAMAM/SWNT LbL film Siqueira Jr. et

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FETs Devices

FETs Devices  Sensitivity of 55 mV/pH – (Ta 2 O 5 : 56-58 mV/pH) 
 Sensitivity of 55 mV/pH – (Ta 2 O 5 : 56-58 mV/pH)  Stability and
Sensitivity of 55 mV/pH – (Ta 2 O 5 : 56-58 mV/pH)
Stability and fast response

Siqueira Jr. et al, Biosens. Bioelectron, 2009

FETs Devices  Sensitivity of 55 mV/pH – (Ta 2 O 5 : 56-58 mV/pH) 

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Catechol Biosensors

Catechol Biosensors -Immobilization of Cl-catechol 1,2 dioxygenase (CCD) and Polyamidoamine G4 Dendrimer (PAMAM) 0.30 10 -2

-Immobilization of Cl-catechol 1,2 dioxygenase (CCD) and Polyamidoamine G4 Dendrimer (PAMAM)

0.30 10 -2 M Catechol 0.25 0.20 0.15 0.10 0.05 10 -11 M Catechol 0.020 0.00
0.30
10 -2 M Catechol
0.25
0.20
0.15
0.10
0.05
10 -11 M Catechol
0.020
0.00
0
20
40
60
80
100
0.015
Time (S)
0.010
10 -11 M Catechol
0.005
Optical Detection
0.000
0
10
20
30
40
ABS at 260 nm

Zucolotto et al., Biosens. and Bioelectron., 21, 1320, 2006.

Catechol Biosensors -Immobilization of Cl-catechol 1,2 dioxygenase (CCD) and Polyamidoamine G4 Dendrimer (PAMAM) 0.30 10 -2

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Catechol Biosensors

Catechol Biosensors 2x10 8x10 1x10 C (F) -7 -7 -8 4x10 -8 0 10 - 10
2x10 8x10 1x10 C (F) -7 -7 -8
2x10
8x10
1x10
C (F)
-7
-7
-8
4x10 -8
4x10
-8
0
0
10 - 10 M Catechol Mili-Q Water 100 Hz - 4 10 - 6 10 -
10 - 10 M Catechol
Mili-Q Water
100 Hz
- 4
10
- 6
10
- 8
10
- 2
10
M
M
M
M
10 - 10 M Catechol Mili-Q Water 100 Hz - 4 10 - 6 10 -
10 - 10 M Catechol Mili-Q Water 100 Hz - 4 10 - 6 10 -
10 - 10 M Catechol Mili-Q Water 100 Hz - 4 10 - 6 10 -
10 - 10 M Catechol Mili-Q Water 100 Hz - 4 10 - 6 10 -
10 - 10 M Catechol Mili-Q Water 100 Hz - 4 10 - 6 10 -

Electrical Detection

2x10 8x10 1x10 C (F) -7 -7 -8 4x10 -8 0 10 - 10 M Catechol
  • PAMAM/CCD

Sensing Units

Bare Electrode

PAMAM/PVS

PAMAM/CCD Sensing Units Bare Electrode PAMAM/PVS
PAMAM/CCD Sensing Units Bare Electrode PAMAM/PVS

Zucolotto et al., Biosensors and Bioelectronics, 21, 1320, 2006.

Catechol Biosensors 2x10 8x10 1x10 C (F) -7 -7 -8 4x10 -8 0 10 - 10

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<a href=E nzyme: Alcohol dehydrogenase (ADH) from yeast (E.C. 1.1.1.1), molecular weight of 141-151 kDa 1500 1250 1000 750 m (ng) 500 250 0 2 4 6 8 10 Number of bilayers Deposition 139.5 ng per bilayer 52.1 ng attributed to ADH. A.C. Perinotto et al. , Thin Solid Films, 2008 nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br " id="pdf-obj-55-2" src="pdf-obj-55-2.jpg">

Enzyme: Alcohol dehydrogenase (ADH) from yeast (E.C. 1.1.1.1), molecular weight of 141-151 kDa

1500 1250 1000 750 m (ng) 500 250 0 2 4 6 8 10 Number of
1500
1250
1000
750
m (ng)
500
250
0
2
4
6
8
10
Number of bilayers

Deposition

139.5 ng per bilayer 52.1 ng attributed to ADH.

<a href=E nzyme: Alcohol dehydrogenase (ADH) from yeast (E.C. 1.1.1.1), molecular weight of 141-151 kDa 1500 1250 1000 750 m (ng) 500 250 0 2 4 6 8 10 Number of bilayers Deposition 139.5 ng per bilayer 52.1 ng attributed to ADH. A.C. Perinotto et al. , Thin Solid Films, 2008 nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br " id="pdf-obj-55-16" src="pdf-obj-55-16.jpg">
<a href=E nzyme: Alcohol dehydrogenase (ADH) from yeast (E.C. 1.1.1.1), molecular weight of 141-151 kDa 1500 1250 1000 750 m (ng) 500 250 0 2 4 6 8 10 Number of bilayers Deposition 139.5 ng per bilayer 52.1 ng attributed to ADH. A.C. Perinotto et al. , Thin Solid Films, 2008 nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br " id="pdf-obj-55-18" src="pdf-obj-55-18.jpg">

A.C. Perinotto et al., Thin Solid Films, 2008

<a href=E nzyme: Alcohol dehydrogenase (ADH) from yeast (E.C. 1.1.1.1), molecular weight of 141-151 kDa 1500 1250 1000 750 m (ng) 500 250 0 2 4 6 8 10 Number of bilayers Deposition 139.5 ng per bilayer 52.1 ng attributed to ADH. A.C. Perinotto et al. , Thin Solid Films, 2008 nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br " id="pdf-obj-55-26" src="pdf-obj-55-26.jpg">

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Pasteurellosis Detection

Pasteurellosis Detection Pasteurellosis: Zoonosis caused by Pasteurella multocida Strategy -Incorporation of Pasteurella multocida membrane protein into

Pasteurellosis: Zoonosis caused by Pasteurella multocida

Strategy

-Incorporation of Pasteurella multocida membrane protein into liposomes.
-Incorporation of Pasteurella multocida membrane protein into
liposomes.

-Proteoliposome immobilization in LbL films.

Pasteurellosis Detection Pasteurellosis: Zoonosis caused by Pasteurella multocida Strategy -Incorporation of Pasteurella multocida membrane protein into
Pasteurellosis Detection Pasteurellosis: Zoonosis caused by Pasteurella multocida Strategy -Incorporation of Pasteurella multocida membrane protein into
Pasteurellosis Detection Pasteurellosis: Zoonosis caused by Pasteurella multocida Strategy -Incorporation of Pasteurella multocida membrane protein into

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Pasteurellosis Detection

Pasteurellosis Detection 100 Hz IgG + - IgG IgG + = IgG anti-P. multocida IgG -
100 Hz IgG + - IgG IgG + = IgG anti-P. multocida IgG - = IgG
100 Hz
IgG +
-
IgG
IgG + = IgG anti-P. multocida
IgG - = IgG anti-sheep erythrocytes
PC1 (73.8%)
PC2 (23.1%)

Zucolotto et al., Analytical Chemistry 79, 2163, 2007.

Pasteurellosis Detection 100 Hz IgG + - IgG IgG + = IgG anti-P. multocida IgG -

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Immobilization of TRB1 TH nuclear receptor for Thyroid hormone detection Substrato de vidro Trilhas de ouro
Immobilization of TRB1 TH nuclear receptor for Thyroid hormone detection
Immobilization of TRB1 TH nuclear receptor for Thyroid
hormone detection
Substrato de vidro Trilhas de ouro Monocamadas Eletrodo de tiol interdigitado Monocamadas de silano
Substrato de vidro
Trilhas de ouro
Monocamadas
Eletrodo
de tiol
interdigitado
Monocamadas
de silano
Immobilization of TRB1 TH nuclear receptor for Thyroid hormone detection Substrato de vidro Trilhas de ouro

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zuco@ifsc.usp.br

[C] = 50,0 nM at 100 Hz Bendo et al., Submitted . nanomedicina.com.br Prof. Dr. Valtencir
[C] = 50,0 nM at 100 Hz Bendo et al., Submitted . nanomedicina.com.br Prof. Dr. Valtencir

[C] = 50,0 nM at 100 Hz

[C] = 50,0 nM at 100 Hz Bendo et al., Submitted . nanomedicina.com.br Prof. Dr. Valtencir

Bendo et al., Submitted.

[C] = 50,0 nM at 100 Hz Bendo et al., Submitted . nanomedicina.com.br Prof. Dr. Valtencir

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Final Remarks nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br
Final Remarks
Final Remarks
Final Remarks nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Biomolecule Analyte Transducer LOD Aplication Glucose oxidase Glucose Amperometric 26  M Soft Drinks Fructose Frutose

Biomolecule

Analyte

Biomolecule Analyte Transducer LOD Aplication
Biomolecule Analyte Transducer LOD Aplication

Transducer

Biomolecule Analyte Transducer LOD Aplication
Biomolecule Analyte Transducer LOD Aplication

LOD

LOD

Aplication

Aplication

Glucose oxidase

Glucose

Amperometric

Glucose oxidase Glucose Amperometric 26  M Soft Drinks

26 M

Glucose oxidase Glucose Amperometric 26  M Soft Drinks

Soft Drinks

Fructose

Frutose

Amperometric

2.4 M

Honey

dehydrogenase

dehydrogenase
dehydrogenase
dehydrogenase

Alcohol

Ethanol

Amperometric

Wine, Beer

dehydrogenase

dehydrogenase
dehydrogenase
dehydrogenase

Glycerol

Glycerol

Absorbance

8 mg/L

Wine

dehydrogenase

dehydrogenase
dehydrogenase
dehydrogenase

Cholesterol

Cholesterol

Fluorescence

70 M

Butter

oxidase

oxidase
oxidase
oxidase

Sulfite oxidase

Sulfite

Sulfite oxidase Sulfite Amperometric 4 ppm Water

Amperometric

Sulfite oxidase Sulfite Amperometric 4 ppm Water

4 ppm

Sulfite oxidase Sulfite Amperometric 4 ppm Water

Water

Monoamino

Histamine

Amperometric

Fish, Meat,

oxidase

oxidase Wine, Beer
oxidase Wine, Beer
oxidase Wine, Beer

Wine, Beer

Acetyl

Pesticides

Termometric

0.2 ng/ml

Fruits,

cholinesterase

cholinesterase Vegetables,
cholinesterase Vegetables,
cholinesterase Vegetables,

Vegetables,

Carboxy

Penicillin G

SPR

5.2 g/kg

Milk

peptidase

 

Sensors and Actuators B 102 (2004) 179194

Biomolecule Analyte Transducer LOD Aplication Glucose oxidase Glucose Amperometric 26  M Soft Drinks Fructose Frutose

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

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Analytical Parameters

Analytical Parameters  Sensitivity (SEN) Increase in the response signal per a specific amount of analyte
  • Sensitivity (SEN)

Increase in the response signal per a specific amount of analyte

  • Limit of Detection (LOD)

Response Signal

 Sensitivity (SEN) Increase in the response signal per a specific amount of analyte  Limit

[C]

Lowest amount of analyte a biosensor can properly recognize

LOD = 3S b /SEN

(Sb = standard deviation for blank measurements*,

SEN = Sensitivity)

  • Limit of Quantification (LOQ)

*When blank analysis gives non-zero deviation

What is lowest amount of analyte a biosensor can quantify?

LOD = 10S b /SEN

Thevenot et al., Pure Appl. Chem., Vol. 71, 12, 2333, 1999. Long et al., Analytical Chemistry, 55, 7, 1983

Analytical Parameters  Sensitivity (SEN) Increase in the response signal per a specific amount of analyte

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Prof. Dr. Valtencir Zucolotto

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Important Parameters

Important Parameters  Specificity, Selectivity How specific a biosensor can be?  Response time Typically 90%
  • Specificity, Selectivity

How specific a biosensor can be?

Response time Typically 90% of steady-state response Reproducibility

Measure of the scatter in consecutive measurements

  • Stability

  • Lifetime

Storage vs operation conditions

  • Reusability

Are the biosensors reusable or disposable?

Thevenot et al., Pure Appl. Chem., Vol. 71, 12, 2333, 1999. Long et al., Analytical Chemistry, 55, 7, 1983

Important Parameters  Specificity, Selectivity How specific a biosensor can be?  Response time Typically 90%

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An Introduction to Ultrathin Organic Films: From Langmuir-- Blodgett to Self — Assembly , <a href=Abraham Ulman , Academic Press (April 28, 1991). Biomaterials Science , 2 Edition : An Introduction to Materials in Medicine by BD. Ratner, A.S. Hoffman, F. J. Schoen, J. E. Lemons Academic Press (7/29/2004). Biosensors: Theory and Applications , by Donald G. Buerk (Author) CRC; 1st edition (August 24, 1995) ISBN-10: 0877629757, ISBN-13: 978-0877629757. Thevenot et al., Pure Appl. Chem. , Vol. 71, 12, 2333, 1999. Long et al., Analytical Chemistry , 55, 7, 1983. nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br " id="pdf-obj-64-2" src="pdf-obj-64-2.jpg">

An

Introduction

to

Ultrathin

Organic

Films:

From

Langmuir--

Blodgett to SelfAssembly, Abraham Ulman, Academic Press (April

28, 1991).

Biomaterials Science, 2 nd Edition : An Introduction to Materials in Medicine by BD. Ratner, A.S. Hoffman, F. J. Schoen, J. E. Lemons Academic Press (7/29/2004).

Biosensors: Theory and Applications, by Donald G. Buerk (Author) CRC; 1st edition (August 24, 1995) ISBN-10: 0877629757, ISBN-13:

978-0877629757.

Thevenot et al., Pure Appl. Chem., Vol. 71, 12, 2333, 1999.

Long et al., Analytical Chemistry, 55, 7, 1983.

An Introduction to Ultrathin Organic Films: From Langmuir-- Blodgett to Self — Assembly , <a href=Abraham Ulman , Academic Press (April 28, 1991). Biomaterials Science , 2 Edition : An Introduction to Materials in Medicine by BD. Ratner, A.S. Hoffman, F. J. Schoen, J. E. Lemons Academic Press (7/29/2004). Biosensors: Theory and Applications , by Donald G. Buerk (Author) CRC; 1st edition (August 24, 1995) ISBN-10: 0877629757, ISBN-13: 978-0877629757. Thevenot et al., Pure Appl. Chem. , Vol. 71, 12, 2333, 1999. Long et al., Analytical Chemistry , 55, 7, 1983. nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br " id="pdf-obj-64-49" src="pdf-obj-64-49.jpg">

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br
nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br
nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br

Muito Obrigado
Muito Obrigado
Muito Obrigado Valtencir Zucolotto zuco@ifsc.usp.br <a href=www.nanomedicina.com.br www.lnn.ifsc.usp.br www.twitter.com/nanomedicina Instituto de Física de São Carlos - USP nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br " id="pdf-obj-66-4" src="pdf-obj-66-4.jpg">
Valtencir Zucolotto zuco@ifsc.usp.br
Valtencir Zucolotto
zuco@ifsc.usp.br

Instituto de Física de São Carlos - USP

Muito Obrigado Valtencir Zucolotto zuco@ifsc.usp.br <a href=www.nanomedicina.com.br www.lnn.ifsc.usp.br www.twitter.com/nanomedicina Instituto de Física de São Carlos - USP nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br " id="pdf-obj-66-16" src="pdf-obj-66-16.jpg">
Muito Obrigado Valtencir Zucolotto zuco@ifsc.usp.br <a href=www.nanomedicina.com.br www.lnn.ifsc.usp.br www.twitter.com/nanomedicina Instituto de Física de São Carlos - USP nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br " id="pdf-obj-66-18" src="pdf-obj-66-18.jpg">
Muito Obrigado Valtencir Zucolotto zuco@ifsc.usp.br <a href=www.nanomedicina.com.br www.lnn.ifsc.usp.br www.twitter.com/nanomedicina Instituto de Física de São Carlos - USP nanomedicina.com.br Prof. Dr. Valtencir Zucolotto zuco@ifsc.usp.br " id="pdf-obj-66-20" src="pdf-obj-66-20.jpg">

nanomedicina.com.br

Prof. Dr. Valtencir Zucolotto

zuco@ifsc.usp.br