AMRg: Applied Molecular Receptors GroupDepartment of Biological Organic Chemistry

IIQAB: Environmental and Chemical Research InstituteJorge Girona, 18-26, 08034-Barcelona, Spain

CSIC: Spanish Council for Scientific Research

CIBER: Bioengenieria, Biomateriales y Nanomedicina:

IQAC: Advanced Chemical Research Institute of CataloniaJorge Girona, 18-26, 08034-Barcelona, SpainIQACDepartment of Chemical and Biomolecular Nanotechnology

Nanobiotechnology

Biotechnology: technological application that uses biological systems, to make or modify products or processes with specific uses in agriculture, food and environmental sciences

and medicine.

Biotechnology

What do we understand by Nanobiotechnology ?

Nanoscience

Nanoscience ( Greek: “nanos” (or Latin: “nanus”) Nano: 10-9 m (scale of atomic diameter) + Science): Study of the atoms, molecules and objects whose size is on the nanometer scale (1-100 nm)

Physic and chemistry is different at the nanometer scale. New properties, not seen on a macroscopic scale, now become important and open the door to a new challenges and possibilities for therapy, diagnostics, microelectronics, environment, etc.

(Bio)Analytical Techniques

Development o analytical techniques based on the molecular recognition event.

Preparation, characterization of the necessary reagents and immunoreagents: Selective receptors of natural (antibodies) and synthetic (molecular imprinted polymers, MIPs) with directed affinity and specificity.

Establishment of analytical protocols based on the use of these reagents.

Synthesis of haptens and bioconjugates: haptenized proteins and enzymes, fluorescent and biotinilated probes, etc.

Organic Chemistry

AMR Group Presentation: Research

Biochemistry

Materials Science

Microelectronics

BioanalyticsAMRg

Nanobiotechnology

Development of biomaterials in which at least one or several of their functional components have nanometric dimensions.

Incorporation of biomolecules on nanostructured functional surfaces of biosensor devices.

Labeling of biomolecules with nanoparticleswith specific optical or/and magnetic properties (quantum dots, noble metal nanoparticles, etc.).

Surface Chemistry

Development of surface derivatization procedures.

Construction of homogeneous, organized, biocompatible and stable functional surfaces to be incorporated into biosensor devices responding to the necessary specificity and detectability of each application

Nanosciences: There is really a NanoWorld?

Nanoscale devices are 100 to 10000 times smaller than human cells.They are similar in size to large biological molecules ("biomolecules") such as enzymes and receptors.Nanoscale devices smaller than 50 nanometers can easily enter most cells, while those smaller than 20

nanometers can move out of blood vessels as they circulate through the body.

The rational design of the receptors is based on a continuous effort for better understanding all the mechanisms involved on the molecular recognition event making use of experimental and theoretical data..

The preparation of the receptors with particular affinity and specificity is made according to the requirements of each specific application.

Design and Development of Selective Receptors of Natural or Synthetic Origin for Their Application on Diagnostic Methods and Devices

AMR Group Presentation: Main Objectives

Amplifier

A/D

Data Acquisition/Display

Transducer

Selective Recognizing Element

Electronic system

a. Catalyticb. Affinity-based

a. Electrochemical biosensorb. Optical biosensorc. Piezoelectric biosensord. Thermometric biosensor

Selective Sensors

a. Natural b. Synthetica. Antibodies b. MIP

Chemical Structure of the Antibodies

Variable region

Constant region

Antibody Binding Site

Variable region

16 nm

12 n

m

Molecular Imprinted Polymers (MIP) as SYNTHETIC Receptors

A) Complexation (Template / Monomers)B) PolimerizationC) Removal of template

Rigid polymer complementary to the ANALYTE, and with affinity for it

•Irgarol, terbutylazine, terbutrin, terbumeton•Atrazine, Simazine•Triazine metabolites (DEHA)•4-Nitrophenol•2,4-Dinitrophenol•2,4,6-Trichloro(bromo)phenol•2,4,5-Trichlo(bromo)phenol

Immunoreagents for Pesticides

• Linear Alkylbenzene Surfactants (LAS)• Sulfophenyl carboxylates (SPCs)• Nonylphenol (NP)•2,4,6-Trichloro(bromo)phenol•2,4,5-Trichlo(bromo)phenol•2,4,6-Trichloro(bromo)anisol

• Sulfonamides•Fluoroquinolones•Tetracyclines•Boldenone•Methylboldenone•Stanozolol•Tetrahydrogestrinene

Immunoreagents for Industrial Pollutants

Immunoreagents for Veterinary Drugs

OHCl

Cl

ClN

N

N

S

NHHN

N

N

N

OH

NHH2N

OH

NO2

NO2

O

OH

N

CO2EtF

NR2N

R2

OOH

NNH

H

OH

C9H19

SO3Na

( )m ( )nCOOH

SO3Na

( )m ( )n

AMRg: Immunoreagents

Detection of pesticides using electrochemical nanobioanalytical platforms

Detection of anabolic androgenic steroids (AAS) byLocalized plasmon resonance (LPR) effect of nobel metal nanoparticles

Nano-Bioanalytic Platforms based on Molecular Recognition

Examples of the investigations at the AMRg

Preparation of receptors with directed selectivity for the analytes of interest. Characterization of the properties of the bioreceptors.Incorporation of the bioreceptors into nanostructureddevices to construct novel functional nanobiomaterials.

Electrochemical NanobioAnalytical Platforms:Prepartion of Nanoparticle-Immunoreagents Hybrid Systems

MNP

100 nm

Anti-Atrazine IgGcovalently coupled to a Magnetic NanoParticle

Au

40 nm

Anti-Atrazine IgG covalently coupled to Gold NanoParticles

10-3 10-2 10-1 100 101 102 1030.00.51.01.52.02.53.03.5

Atrazine (nM)-I(μ

A)

HRP

ne-

HQHRPred OX

ox red

H2O H2O2

B

Wash stepusing a magnet

Amperometricmeasures

D

A C

IC50 = 3 ng L-1

LOD= 0.3 ng L-1

E. Zacco, M. I. Pividori, S. Alegret, R. Galve and M.-P. Marco. Anal. Chem., 78, 1780-1788, 2006

Amperometric MagnetoSensor for Atrazine Analysis N

N

N

Cl

NH NH

Atrazine

10-3 10-2 10-1 100 101 102 1030123456789

Juice 1:5PBST

MP-Tosyl 10μg; 2b-HRP=0.05μg/mL

Atrazina (nM)

-I(μ

A)

Amperometric Magneto ImmunosensorAnalysis of Orange Juice Samples

C

Atrazine

N

N

N

Cl

NHHN

Magneto SENSORMagneto SENSOR

E. Zacco, M. I. Pividori, S. Alegret, R. Galve and M.-P. Marco. Anal. Chem., 78, 1780-1788, 2006

NH2

CH2

CH2

CH2

SiO O O

NH2

CH2

CH2

CH2

SiO O O

NH2

CH2

CH2

CH2

SiO O O

NH2

CH2

CH2

CH2

SiO O O

NH2

CH2

CH2

CH2

SiO O O

NH2

CH2

CH2

CH2

SiO O O

NH2

CH2

CH2

CH2

SiO O O

NH2

CH2

CH2

CH2

SiO O O

10-2 10-1 100 101 102 103 104 1050.0

0.1

0.2

0.3

[Atrazine, μg/L]

pF

200 um 50um 10 um 5 um

Impedimetric and Conductimetric Sensor for Atrazine Analysis

Sensors and Actuators, submitted

NH

CH2

CH2

CH2

SiO O O

NH

CH2

CH2

CH2

SiO O O

NH

CH2

CH2

CH2

SiO O O

Au

NH

CH2

CH2

CH2

SiO O O

NH

CH2

CH2

CH2

SiO O O

NH

CH2

CH2

CH2

SiO O O

Au

NH

CH2

CH2

CH2

SiO O O

NH

CH2

CH2

CH2

SiO O O

NH

CH2

CH2

CH2

SiO O O

Au

NH

CH2

CH2

CH2

SiO O O

NH

CH2

CH2

CH2

SiO O O

NH

CH2

CH2

CH2

SiO O O

Au

NH

CH2

CH2

CH2

SiO O O

NH

CH2

CH2

CH2

SiO O O

NH

CH2

CH2

CH2

SiO O O

Au

NH

CH2

CH2

CH2

SiO O O

NH

CH2

CH2

CH2

SiO O O

NH

CH2

CH2

CH2

SiO O O

Au

NH

CH2

CH2

CH2

SiO O O

NH

CH2

CH2

CH2

SiO O O

NH

CH2

CH2

CH2

SiO O O

Au

glass

Nor

mal

ised

Extin

ctio

n

540 560 580Wavelength (nm)

1.0

0.9

600520

0.8

0.7

Stanozolol-BSA

Stanozolol

Anti-stanozolol

LPR sensor (Localised plasmon resonance)

Shift

Biosens. Bioelectron. 2006, 21, 1345

LPR sensor (Localised plasmon resonance)

As147/B_BSA (LPR)IC50 μg/L 32

Slope -0.98

6

0.98

LOD μg/Lr2

NHN

OH

H

Biosens. Bioelectron. 2006, 21, 1345

Acknowledgements

EC Information Socity and Technology:GOODFOOD FP6-IST-2003-508774

EC, Nanotechnology and nanosciences, ELISHA FP6-NMP2-CT-2003-505485

MCyT, Recursos y Tecnologias Agroalimentarias:PANOPTES AGL2005-07700-C06-01NANOBIOMOL NAN2004-09415-C05-02NANOBIOMAG NAN2004-09195-C04-04