folic acid grafting to ag nanoclusters-silica thin films
DESCRIPTION
FOLIC ACID GRAFTING TO Ag NANOCLUSTERS-SILICA THIN FILMS. S. Ferraris , S. Perero, E. Vernè, L. Rimondini, E. Battistella and M. Ferraris. Cancer cells differ from heathy one for growth control, morphology, cell-to-cell interactions, membrane properties, cytoskeletal structure, - PowerPoint PPT PresentationTRANSCRIPT
FOLIC ACID GRAFTING TO Ag FOLIC ACID GRAFTING TO Ag NANOCLUSTERS-SILICA THIN NANOCLUSTERS-SILICA THIN
FILMSFILMSS. FerrarisS. Ferraris, S. Perero, E. Vernè, L. Rimondini, E. , S. Perero, E. Vernè, L. Rimondini, E.
Battistella and M. FerrarisBattistella and M. Ferraris
THE PROBLEMTHE PROBLEM
Cancer cells detection at the Cancer cells detection at the early developement stage.early developement stage.
Cancer cells differ from heathy one Cancer cells differ from heathy one forfor
growth control, growth control,
morphology, morphology,
cell-to-cell interactions,cell-to-cell interactions,
membrane properties, membrane properties,
cytoskeletal structure, cytoskeletal structure,
protein secretion, protein secretion,
vascularizationvascularization
gene expressiongene expression
… …
CANCER CELLS CANCER CELLS RAPID DIVISION RAPID DIVISION FOLIC ACID NEED FOLIC ACID NEED FOR DNA SYNTHESISFOR DNA SYNTHESIS
AIM of THE AIM of THE WORKWORK
Ag
Ag
Ag
Ag
Ag
Silver nanoclusters-Silver nanoclusters-silica composites thin silica composites thin
filmsfilms
FA FA FA
Ab
sorb
an
ce [
a.u
]Wavelength [nm]
Characteristic SPR absorption
New SPR absorption
S. Ferraris, S. Perero, E. Vernè, E. Battistella, L. Rimondini; M. Ferraris “Surface functionalization of Ag-nanoclusters-silica composite films for biosensing accepted to Materials Chemistry and Physics
SILICASILICA AgAg
ArAr ArAr
SUBSTRATESUBSTRATE
SUBSTRATESUBSTRATE
SILVER NANOCLUSTERS-SILICA LAYERSILVER NANOCLUSTERS-SILICA LAYER
THE SILVER NANOCLUSTERS-SILICA FILMTHE SILVER NANOCLUSTERS-SILICA FILM
S. Ferraris, S. Perero, E. Vernè, E. Battistella, L. Rimondini; M. Ferraris “Surface functionalization of Ag-nanoclusters-silica composite films for biosensing accepted to Materials Chemistry and Physics
SPR absorptionSPR absorption
SPR absorptionSPR absorption
THE SILVER NANOCLUSTERS-SILICA FILMTHE SILVER NANOCLUSTERS-SILICA FILM
AS DEPOSITEDAS DEPOSITED TT 450°CTT 450°C
100 nm 100 nm
S. Ferraris, S. Perero, E. Vernè, E. Battistella, L. Rimondini; M. Ferraris “Surface functionalization of Ag-nanoclusters-silica composite films for biosensing accepted to Materials Chemistry and Physics
Acetone and water washings
Surface pre-treatment cleaning and hydroxyls exposition
OHOH OH
Ag Ag Ag
THE GRAFTING PROCEDURETHE GRAFTING PROCEDURE
Silanization
3-aminopropyltriethoxysilane (APTES)
Functional group
=
Amino-groupAg
Ag
Ag
NH2
Si
NH2
Si
Folic Acid (FA) Grafting
DCC
FA
Ag
Ag
Ag
AP
TE
SFA
AP
TE
SFA
AP
TE
SFA
S. Ferraris, S. Perero, E. Vernè, E. Battistella, L. Rimondini; M. Ferraris “Surface functionalization of Ag-nanoclusters-silica composite films for biosensing accepted to Materials Chemistry and Physics
SILANIZATIONSILANIZATION
0
10
20
30
40
50
60
70
80
90
AsDep TT450
Co
nta
ct
an
gle
[°]
Washed
Silanized
AS DEPOSITEDAS DEPOSITED TT 450°CTT 450°C
Atomic %
2,40N
4,46,3Ag
17,417,8Si
35,528,2C
40,347,6O
SilanizedWashedElement
XPS survey analysisXPS survey analysis XPS survey analysisXPS survey analysis
Atomic %
4,50N
2,43Ag
18,422,3Si
28,117,3C
46,657,5O
SilanizedWashedElement
Cou
nts
Binding Energy [eV]
a
NH2
Cou
nts
Binding Energy [eV]
bNH2
S. Ferraris, S. Perero, E. Vernè, E. Battistella, L. Rimondini; M. Ferraris “Surface functionalization of Ag-nanoclusters-silica composite films for biosensing accepted to Materials Chemistry and Physics
FOLIC ACID GRAFTINGFOLIC ACID GRAFTING
AS DEPOSITEDAS DEPOSITED TT 450°CTT 450°C
Atomic %
7,92,40N
3,84,46,3Ag
14,817,417,8Si
38,935,528,2C
34,640,347,6O
FA+DCCSilanizedWashedElement
XPS survey analysisXPS survey analysis
Atomic %
5,84,50N
0,82,43Ag
14,718,422,3Si
41,428,117,3C
37,446,657,5O
FA+DCCSilanizedWashedElement
XPS survey analysisXPS survey analysis
NH
Si
NH2
Si
NH2
Si
Ag AgAg
Ag Ag
G006C9_1.SPE: Politecnico
2010 Feb 3 Al mono 25.6 W 100.0 µ 45.0° 23.50 eV 1.2802e+003 max 3.03 min
C1s/Area1/1 (BKG_S)
278280282284286288290292294296298-200
0
200
400
600
800
1000
1200
1400
G006C9_1.SPE
Binding Energy (eV)
c/s
Pos. Sep. %Area283.64 0.00 58.42284.73 1.09 31.26286.41 2.77 10.32
Cou
nts
Binding Energy [eV]
C-C
NHCO
COOH
H0002_1.SPE: Politecnico
2010 Feb 3 Al mono 25.6 W 100.0 µ 45.0° 23.50 eV 1.3465e+003 max 6.73 min
C1s/Area1/1 (BKG_S)
278280282284286288290292294296298-200
0
200
400
600
800
1000
1200
1400
H0002_1.SPE
Binding Energy (eV)c/
s
Pos. Sep. %Area283.64 0.00 49.85284.26 0.62 44.69286.69 3.05 5.45
Binding Energy [eV]
Cou
nts
C-C
NHCO
COOH
G0002_1.SPE: Politecnico
2010 Feb 3 Al mono 25.6 W 100.0 µ 45.0° 23.50 eV 3.5929e+002 max 3.37 min
N1s/Area1/1 (BKG_L)
390392394396398400402404406408410412
-50
0
50
100
150
200
250
300
350
400
G0002_1.SPE
Binding Energy (eV)
c/s
Pos. Sep. %Area398.32 0.00 78.10399.93 1.62 21.90
Cou
nts
Binding Energy [eV]
NH2
NHCO
H0002_1.SPE: Politecnico
2010 Feb 3 Al mono 25.6 W 100.0 µ 45.0° 23.50 eV 2.6197e+002 max 6.73 min
N1s/Area1/1 (BKG_L)
390392394396398400402404406408410412
0
50
100
150
200
250
300H0002_1.SPE
Binding Energy (eV)
c/s
Pos. Sep. %Area398.39 0.00 80.65400.01 1.62 19.35
Cou
nts
Binding Energy [eV]
NH2
NHCO
S. Ferraris, S. Perero, E. Vernè, E. Battistella, L. Rimondini; M. Ferraris “Surface functionalization of Ag-nanoclusters-silica composite films for biosensing accepted to Materials Chemistry and Physics
Ag
Ag
Ag
Ag
Ag
FA FA FA
PRIMARY ANTIBODY PRIMARY ANTIBODY (FOLATE 8/33)(FOLATE 8/33)
SECONDARY ANTIBODY (SECONDARY ANTIBODY (Rabbit Rabbit polyclonal to Mouse IgG polyclonal to Mouse IgG (HRP) (HRP) ))
ELISA TEST (*)
(*) Eastern Piedmont University A. AvogadroREACTION WITH REACTION WITH
CROMOGENEOUS COMPOUND CROMOGENEOUS COMPOUND AND UV-MEASUREMENT OF AND UV-MEASUREMENT OF
ABSORBANCEABSORBANCE
S. Ferraris, S. Perero, E. Vernè, E. Battistella, L. Rimondini; M. Ferraris “Surface functionalization of Ag-nanoclusters-silica composite films for biosensing accepted to Materials Chemistry and Physics
Folic acid grafted Ag-Folic acid grafted Ag-nanoclusters-silica thin nanoclusters-silica thin
films are able to films are able to selectively bind the selectively bind the
proper antibodyproper antibody
CONCLUSIONS …CONCLUSIONS …
Silver nanoclusters-silica composite films Silver nanoclusters-silica composite films have been effectively silanized with 3-have been effectively silanized with 3-aminopropyltriethoxysilaneaminopropyltriethoxysilane
Folic acid has been successfully grafted to Folic acid has been successfully grafted to silanized films via carbodiimide activationsilanized films via carbodiimide activation
The characteristic SPR absorption is The characteristic SPR absorption is maintained after functionalizationmaintained after functionalization
FA-functionalized films are able to selectively FA-functionalized films are able to selectively recognize the proper antibodyrecognize the proper antibody
… … AND FUTURE DEVELOPMENTSAND FUTURE DEVELOPMENTS
SPR absorption measurement in presence of SPR absorption measurement in presence of the antibody will be performed in humid the antibody will be performed in humid environment in order to evaluate the shift due environment in order to evaluate the shift due to selective bindingto selective binding
Recognition experiment with marked healthy Recognition experiment with marked healthy and cancerous cells will be performed and cancerous cells will be performed
ThankThanks!s!