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