nanomaterials for biosensors technology · part b slc ~50 nm sln ~50 nm polymer micelle ~150 nm...
TRANSCRIPT
Tuesday, 03 March 2015 – 13.15 – 15.00 – Room E330
TFYA62 Course
Nanomaterials
for biosensors technology
Ashutosh Tiwari, PhD, Doc Associate Professor
Biosensors and Bioelectronics Centre
2
What is nanomaterials?
• Nanomaterials, size in between 1-100 nm are tools to
work in nanotechnology.
• Inorganic: nanopowders, nanoparticle dispersions, and
surface-functionalized nanoparticles.
• Organic: Carbon nanotubes, graphene,
fullerenes, dendrimers, dendrons and hyperbranched
polymers.
• Composite: Nanoclays and core-shell inorganic-organic
hybrids.
3
Advantages of nanomaterials
• High surface area (capacity)
• Well defined structure
• High reactivity
• Easy dispersability
• Readily tailored for application in
• different environments
• Chemistry/materials developed for remediation
processes are readily tailored to biosensing/detection
4
Nano-assembly
Sci China Chem, 55, 2012.
5
Examples of nanostructured
materials
Au and Ag nanoparticles
Dendrimer
Fullerene C60
(0D
)
6
Size-dependent optical properties of quantum dots Band gap energies
Absorption spectra
Procedures for synthesis of colloidal quantum dots Synthesis of quantum dots in reverse micelles
Synthesis of quantum dots in aqueous media
Hot-matrix synthesis of quantum dots
Types of semiconductor quantum dots Binary quantum dots
Alloyed quantum dots
Core/shell quantum dots: “type-I”
Core/shell quantum dots: “type-II”
Quantum dot/quantum well nanocrystals
Transition-element-doped quantum dots
Surface functionalization of quantum dots
Self-assembly of colloidal quantum dots
Quantum dots
7
450 500 550 600 650 700 750 8000
1
2
Wavelength (nm)
Abso
rpti
on (
a.u
.)
0
1
2
PL
Inte
nsity
(a.u
.)
3.5 nm
3.8 nm
4.0 nm
4.5 nm
(a)
(b) (c)
Small LargeSizes of Quantum Dots
(a) Photoluminescence of CdSe QD samples exposed to ultraviolet light. (b) Absorption and
PL spectra of CdTe QDs with varying sizes as indicated for each sample. (c) TEM image of a
film composed of spherical CdTe QDs. The inset in (c) is a selected area electron diffraction
(SAED) pattern from the QD film.
CdSe quantum dots
8
Popular 1D & 2D nanomaterials
Carbon nanotube
Graphene
Au nanorod
9
2D and 3D nanostructures
Thin film 3D assembly
100 nm
10
Functional nanomaterials
• Polymers and composites, one of emerging frontiers in materials science and biomedical studies, deal with induced conformational changes in their structures.
• Smart/and intelligent materials are being formulated in response to the environmental changes, typically with temperature, pH, electricity and magnetism etc.
• The intelligent materials could modulate their conformations and/or structures in these physical environments without causing biological and environmental harm, which thereby make them mainly attractive to biosensor, diagnosis, target-specific molecular recognition, metabolic control mechanisms and drug delivery.
11
Composite or
miniaturize
FORM:
INTELLIGENCE
ADADAPTABILITY
RESPONSIVENESS
DDS In vivo with smart properties:
Biodegradable form
Encapsulation ratio
Polymer
Biocompatibility
Self-assembly or ‘hard’ synthesis
Strategies
Nanofabrication:
•DDS
•Porous and hard particle
•SEDDS, SMEDDS
•Polymer and amphiphile self-assembly
Environment:
Stimuli & responsiveness -
T, t, pH, I, Eh, M+,
reductase enzyme
MIMIC?
NOVEL?
Smart methods:
Biosensor use?
Biomaterials (novel)
key:
T = temperature, t = time,
I = ionic strength, Eh = redox status,
M+ =metal ion sensitivity, e.g. Ca2+
-phenomenological
representation of bottom-up
materials design
considerations.
-the form, environment and
nanofabrication method are
important factors in
production of a smart
technology that is variable in
structure and its use.
Designing strategy
Tiwari, A and Kobayashi, H. (Eds.) In Responsive materials and methods. Wiley, USA 2013.
12
Polymeric nanomaterials
Amphiphilic blocks consist of hydrophilic and hydrophobic chains. They
can self-assemble at the polymeric architectonic.
The synthesis of such structures could be accomplished by living anionic
polymerization, controlled/”living” radical polymerization (CRP) techniques
for example nitroxide-mediated polymerization (NMP), atom-transfer
radical polymerization (ATRP) or reversible addition-fragmentation chain
transfer (RAFT) polymerization.
The cross linking of amphiphilic polymers mainly in aqueous media – as
biomedical applications are targeted- together with the stabilization of
the self-assembled supramolecular structures by crosslinking.
The crosslinking of the self-assembled structure by the introduction of
covalent bonds between chains simultaneously or after assembly is
an efficient way to stabilize such dynamic supramolecular systems.
The stability of supramolecular structures strongly depends on the
nature and length of the polymers, the solvent and the temperature.
Tiwari, A, Kobayashi, H. Turner, A.P.F. (Eds.) In Biomedical materials and diagnostic devices. Wiley, USA 2012.
13
Porous or impenetrable
SOLID or GEL
(50-200 nm)
Active is encapsulated in core
and/or in surface layering depending on log P
Part A
Part B
SLC
~50 nm
SLN
~50 nm
Polymer
Micelle
~150 nm
Nano-
sphere
~150 nm
Liposome
~25 nm
Micro-
emulsion,
Micelle
<10 nm
Quantum
-dot
~nm
Pro-drug
<5 nmL
ipid
Lip
id
Lip
id
Lip
id
PL
GA
, P
LA
,
PC
L
Ph
osp
ho
lipid
Su
rfa
cta
nt
Ch
em
ica
l
co
nju
ga
te
Fine
Emulsion
~400 nm
Lip
id
Hollow or
filled
Use in other
systems
such as tablets
or soft gelatin
capsules
Conjugated
antibody
Stimulus
modifier
Simple or
Polymer
coated
Drug +
linker+
particle
carrier
Pickering
Emulsion
Nano-
Emulsion
Formulation materials:
Natural, chemically-derivatized
Synthetic-biocompatible
LbL or
Adsorbate or
Electrostatic
deposition
Implants:
Prosthetics &
mimetic
Designs of polymers, particle
surfaces and interfaces, soft
nanotechnology products and
materials self-assembly- A flow
chart
Part A - forms such as solid lipid
nanoparticles and solid lipid
capsules, their size range and key
excipient type
-part B indicates the subtlety of
modification that is open to
theranostics scientists
Fabrications and
processing
Tiwari, A.; Tiwari, A. (Eds.) In Nanomaterials in drug delivery, imaging, and tissue engineering. Wiley, USA 2013.
14
Self-assembly Morphological determination of assemblies of amphiphilic blocks
The design of the copolymer at the
molecular level, i.e. the nature of the
blocks, the blocks length and the
chain architecture directly impact the
assembly and morphology.
The type of structure formed is due to the inherent curvature of the molecule, which
can be estimated through calculation of its dimensionless packing parameter.
Blanazs, A.; Armes, S.P.; Ryan, A.J. (2009) Self-assembled block copolymer aggregates: From micelles to vesicles and their biological applications.
Macromol. Rapid Commun. 30, 267–277.
p = v/aolc p = packing parameter
v = volume of hydrophobic chains
ao = optimal area of head group
lc = length of hydrophobic tail
15
Electrospun nanofibers
In the electrospinning process, the
polymer fibers-mat can be
extruded under an anode spinneret
with the electric force to grounded
collector.
- Concentration of polymer solution
- Molecular weight of polymer
- Humidity of electro spinning chamber
- Solvent of solution
- fixed voltage, flow rate and distance
from needle to ground.
16
17
Core-shell nanocomposite
CdSe
Radial distance
Po
ten
tial
EV
EC
Core
organic
molecule
Core/Shell (type-I)
CdSe
Core/Shell (type-II)
CdSe
ZnS ZnTe
CdS
HgS
CdS
Quantum Dot/Quantum Well
Ye
Yh
Illustrations of various types of QDs (top), and potential energy diagrams for the
respective QDs’ electrons and holes (bottom). The dashed and solid black lines
illustrate the electron (Ye) and hole (Yh) wavefunctions, respectively.
18
Polymer vesicles
Formation of polymer vesicles by
simultaneous chain growth and
self-assembly via controlled
radical aqueous emulsion
polymerization
Chem. Commun., 2009, 2887-2889
19
Micelle-vesicle
Reversible light-induced micellization and micelle-vesicle
transition of hydrogen bonded polymers
Angew. Chem. Int. Ed. 2006, 45, 3846-3850
20
Thermo-sensitive nanogel
XXX T > LCST
XX
X
Mn > critical Mn
M1 = hydrophilic monomer, whose polymer possesses a LCST
R = bifunctional monomer
R• = initiator of radical polymerization
hydrophilic polymer
(PEO)
thermosensitive polymer
(PDEAAm)
MacroRAFT
agent
T > LCST
M1, M2, R•
Chain growth,
assembly and
crosslinking
M1, M2, R• T < LCSTM1, M2, R•
XXX
XXX T > LCST
XX
X
Mn > critical Mn
M1 = hydrophilic monomer, whose polymer possesses a LCST
R = bifunctional monomer
R• = initiator of radical polymerization
hydrophilic polymer
(PEO)
thermosensitive polymer
(PDEAAm)
MacroRAFT
agent
T > LCST
M1, M2, R•
Chain growth,
assembly and
crosslinking
M1, M2, R• T < LCSTM1, M2, R•
Synthesis pathway to thermo-sensitive nanogel particles:
simultaneous chain growth, assembly and crosslinking.
Langmuir 2009, 25, 5258–5265
21
Biomolecular interactions
-nanoparticles interacting with proteins,
membranes, cells, DNA and organelles
-establish a series of nanoparticle/biological
interfaces
Nel, A.E. et al. (2009) Understanding biophysicochemical interactions at the nano–bio interface. Nature Materials, 8, 543-557.
- depends on colloidal forces as well
as dynamic biophysicochemical
interactions
22
Hyperbranched polyester functionalized
gold nanoparticles
+
Drop casting EDC/NHS
(A)
(B)
ITO coated
glassH40-Au/ ITO
Carboxyl gold nanoparticle
(Au-COOH)
Amine functionalized Boltorn® H40
(Boltorn® H40-NH2)
Amine functionalized hyperbranched gold (H40-Au) nanoparticle
Urs/H40-Au/ ITO
DDC/NHS
Au NPs H40 -NH2
-COOH Urs-CO-NH-
Talanta 78, 1401–1407, 2009
Fictionalization of gold
nanoparticles with
hyperbranched polyester was
conducted via a two-step
procedure.
gold nanoparticle was
functionalized with mercapto
propionic acid (Au–COOH).
amine functionalized
hyperbranched Boltorn H40
(H40–NH2) was covalently
grafted onto Au–COOH
nanoparticles using N,N-
dicyclohexylcarbodiimide
(DCC) and
Nhydroxysuccinimide (NHS)
mediated reaction
23
Nanofibers - sensor fabrication
Schematic diagram of the fabrication process for polyelectrlyte reactor on the Pt-disc electrode: (a)
Pt-disc electrode coated with PSA/PSSA electrospun fibes-mat, (b) self-assembly of ABBA upon
PSA/PSSA electrospun fibes-mat, (c) interaction of glucose with ABBA over the electrode surface
during glucose sensing, and (d) Formation of boronic ester (VII) via the reaction of 3-aminobenzene
boronic acid (VI) and glucose at physiological pH.
Talanta, 82, 1725, 2010.
Pt-electrode
-+
+
-+
-
-
+ -
- ++
-
--
-
-
-
+
-+
++
+
+-
+
Pt-electrode
-+
+
-+
-
-
+
--+
-
--
-
-
-
+
-
++
+
+
-+
-+
+
++
-
-
+
--+
-
--
-
-
-
+
-
++
+
+
- +
z-
+ -
+-
+ -
+
BOH
OH
NH2
BOH
OH
NH2 RR
OHOH
R
R
O
O
B-
OH
NH2
+ +
--Pt-electrode
a b c
Sensing
Adsorption
PSA/PSSA electrospun
fibers-matABBA
Boronic ester
with glucose
Glucose
B
OH
OH
NH2
RR
OHOH
R
RO
OB-
OH
NH2
VI VIII
pH 7.4
B-
OH
OH
NH2
OHOH
-
+ 2H2O
VII
d
24
Tunable biosensors
ZnOnanoparticles
CHIT-g-PVAL matrix
ZnO/CHIT-g-PVAL /ITOnanocomposite electrode
GOD/ZnO/CHIT-g-PVAL /ITOnanocomposite bioelectrode
GOD
Spin coating+
(a) (b)
ITO
ITO
Schematic illustration of (a) fabrication of ZnO/CHIT-g-PVAL core-shell nanocomposite electrode,
and (b) immobilization of GOD onto core-shell nanocomposite electrode.
i) ZnO nanoparticles enhances the sensitivity of bioelectrode, ii) pH responsive, high swelling
behavior of core-shell nanocomposite film which provides small surface reaction zone for
interferences and good impulse propagating materials for glucose biosensing.
Shukla, S.K., Deshpande, S.R., Shukla, S.K., Tiwari, A. Fabrication of a tunable glucose biosensor based on zinc oxide/chitosan-graft-poly
(vinyl alcohol) core-shell nanocomposite. Talanta, 99, 283–287.
25
Template-directed hierarchical self-assembly of
graphene based ultrasensitive biosensors
-functionalised graphene
as a fundamental building
block to obtain
hierarchically ordered
graphene-enzyme-
nanoparticle hybrid
structures for new
electrode materials
-hybrid structures provide
more sensitive and
efficient electrochemical
sensors, biosensors and
catalytic electrodes.
Parlak, O., Tiwari, A., Turner, A. P. F., Tiwari, A. (2013) Template-directed hierarchical self-assembly of graphene based
hybrid structure for electrochemical biosensing. Biosensors and Bioelectronics, 49, 53-62.
26
On/off-switchable zipper-like
bioelectronics on a graphene interface
Adv. Mater. 2014, 26, 482–486.
-smart and flexible
bioelectronics on
graphene: emerged a
new frontier in the
field of biosensors
-biodevices suffer
from a lack of control
and regulation
-on/off-switchable
zipper-like graphene
interface: manipulate
biomolecular interactions
27
MRI-visual order–disorder micellar
nanostructures
Fabrication of the MRI visual SPION@PS-b-PAADOx/ FA cancer nano-theranostics module.
Adv. Healthcare Mater, 4, XXX, 2014.
- SPIONs: Superparamagnetic iron oxide nanoparticles
- SPION@PS-b-PAADOx/ FA: amphiphilic poly(styrene)- b -poly(acrylic acid)-doxorubicin with folic acid
(FA) surfacing
Composite-DOx/FA
Recognition
Composite-DOx/FA
Dissemble
Drug release
Composite-DOx/FA
Disintegration
Tumor
microenvironment
Acidic pH
28
1) ChOx and ChEt
2) EDC/NHS
Integrated self-reporting
nanobiosensing
Gold nanorods with a monolayer of enzymes-conjugated
amphiphilic block copolymer for NIR based cholesterol
nanobiosensing
29
On/Off-switchable immunosensors
Deshpande, S., Turner, A.P.F. and Tiwari, A. (2012). Label-free nano immunosensor for diagnosis of cardiac injury based on localised surface plasmon
resonance. Label-free Technologies, 1-3 November 2012, Amsterdam, Elsevier.
Gold nanorod-based plasmonic biosensing displayed much higher selectivity and
sensitivity in the picograms/mL range for trace analytes
b) Antibody conjuagtedAuNR
c) AuNR with PNIPAAm
d) Ab-Ag interaction at 25OC
e) Ab-Ag interaction at 37OC
Thermo-switching behaviour
AntibodyTroponin (Ab)
Antigen
Troponin T (Ag)Poly(N-isopropylacrylamide)(PNIPAAm)
a) Gold nanorods(AuNR)
Schematic
illustrations (a-e)
of fabrication
steps of smart
nano-
immunosensor
and regeneration
of sensor at
room
temperature.
30
(a) Troponin T-PNIPAM coil complex and (b) Troponin T- PNIPAM globular complex
(a) (b) -Calculations show
temperature dependence
of secondary structures
namely coil and globular
of PNIPAM which in turn
modulates the interaction
between the antibody
and troponin-T in a
temperature responsive
way.
Electrostatic energy, van der Waals energy, Internal energy, Gas phase energy, non-polar contribution to the solvation free
energy, electrostatic contribution to the solvation free energy, sum of non-polar and polar contributions to solvation, sum of the
electrostatic solvation free energy and electrostatic energy and final estimated binding free energy.
Binding Free Energy of Complexes
31
High-throughput Electrocatalytic
Nano-bioreactors
Two-dimensional gold-tungsten disulphide bio-interface
Parlak, O.; Seshadri, P.; Lundström, I.; Turner, A.P.F; Tiwari, A. Advanced Materials Interfaces, DOI: 10.1002/admi.201400136, 2014.
32
TEM images of WS2 nanosheets
Nano-bioreactors
33
Electrocatalytic Activity of Nano-bioreactors
Amperometric responses (a) and the calibration curves (b) for the sensing of H2O2 with
WS2/HRP and WS2/Au NPs/HRP electrodes in 0.1 M PBS at 0.5 V applied potential vs.
Ag/AgCl.
34
For more reading
http://eu.wiley.com/WileyCDA/W
ileyTitle/productCd-
1118773519.html
35
36
Ashutosh Tiwari, PhD, Docent
Associate Professor and Group Leader
Smart Materials and Biodevices
Room M312
Biosensors and Bioelectronics Centre
IFM-Linköpings universitet
581 83 LINKÖPING, Sweden
Tel: (+46) 013-28 2395
Fax: (+46) 013-13 7568
Web: https://www.ifm.liu.se/applphys/biosensors-and-
bioelectro/group-members/ashutosh-tiwari/
You can contact me-
www.liu.se Thank you!