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Article Nanoscale SI - Royal Society of Chemistry

IntégréTéléchargement
Electronic Supplementary Material (ESI) for Nanoscale.
This journal is © The Royal Society of Chemistry 2016
SupportingInformation
Compactquantumdot-antibodyconjugatesforFRET
immunoassayswithsubnanomolardetectionlimits
LuciaMattera,1-3ShashiBhuckory,4K.DavidWegner,4§FabioAgnese,1,5Christophe
Lincheneau,1-3TimSenden,1-3DavidDjurado,1-3LoïcJ.Charbonnière,6NikoHildebrandt,4*
PeterReiss1-3*
1
Univ.GrenobleAlpes,INAC-SyMMES,F-38054GrenobleCedex9,France
2
CEA,INAC-SyMMES,LaboratoireSTEP,17ruedesMartyrs,F-38054GrenobleCedex9,France
3
CNRS,SPrAM,F-38054GrenobleCedex9,France
4
NanoBioPhotonics (nanofret.com), Institut d'Electronique Fondamentale, Université Paris-Sud, Université
Paris-Saclay,CNRS,91405OrsayCedex,France
5
CEA,INAC-MEM,LEMMA,17ruedesMartyrs,F-38054GrenobleCedex9,France
6
Institut Pluridisciplinaire Hubert Curien (IPHC), UMR 7178 CNRS/UdS, Laboratoire d'Ingénierie Moléculaire
Appliquéeàl'Analyse(LIMAA),ECPM,25,rueBecquerel,67087StrasbourgCedex2,France
§
Presentaddress: CEA,INAC-SyMMES,17ruedesMartyrs,F-38054GrenobleCedex9,France
*Correspondingauthors:niko.hildebrandt@u-psud.fr,peter.reiss@cea.fr
S1
SI-1: UV-vis and PL spectra of InPZnS/ZnSe/ZnS QDs (a), CdSe/ZnS – QD605 (b), CdSe/ZnS - QD705 (c) in
chloroform (black), after functionalization with penicillamine (red) and with the bifunctional ligand Mal1 in PBS
1X buffer.
SI-2: TEM images and size distributions of CdSe705-Pen (left), InP530-Mal1 (middle) and CdSe605-Pen (right).
S2
SI-3: Size distribution of QD705 after each functionalization step (Pen-capping: green; Mal1-functionalization:
red; F(ab)-conjugation: blue) obtained by DLS measurements. The average hydrodynamic diameter d (nm) of
each measurement is given in the legend.
SI-4: Gel electrophoresis characterization of the sample InP530 (a, b), QD705 (c) and QD605 (d) at different
stages of functionalization was carried out on 1% agarose gel in 25 mM Hepes buffer at 100 V on a RunOne
System. Prior to gel electrophoresis, to each sample a solution corresponding to 20% of the sample volume and
containing Orange G and 30% glycerol in 6% loading buffer was added. For visualization, the gel was placed on
a UV transilluminator, and an image was captured with a Gel Doc XR system (Bio-Rad, Hercules, CA). In the
case of InP-based QDs (a-b) the images shown here were acquired after 10 min while for the Cd-based QDs
after 20 min. The label a, c and d refers to the sample functionalized with Pen and Mal1, while b refers to the
sample functionalized with Mal1 and bioconjugated with F(ab).
S3
!
Fab
s86-QD705-Fab
3900
3600
3300
3000
2700
2400
2100
1800
1500
1200
Weavenumber cm-1
SI-5: FTIR characterization showing the successful functionalization of the QDs with the Mal1 ligand and
subsequent conjugation of the fragmented antibody F(ab). Left: FTIR spectra of Mal1 (red), InPZnS/ZnSe/ZnS
QDs functionalized with Pen (blue) and post-functionalized with Mal 1 (green); right: FTIR spectra of F(ab) and
of CdSe/ZnS QD705 after F(ab) conjugation.
L4Tb emission
0,05
InP530 absorption
4,5E5
3,0E5
0,03
0,02
1,5E5
rel. PL intensity
ε / M-1.cm-1
0,04
0,01
0,0
500
550
600
650
700
0,00
Wavelength / nm
2,0E6
L4Tb emission
0,05
6E6
QD 605 absorption
0,04
1,0E6
0,02
5,0E5
0,04
0,03
3E6
0,02
2E6
0,01
0,01
1E6
0,0
450
500
550
600
Wavelength / nm
650
0,00
700
0,05
4E6
ε / M-1.cm-1
0,03
L4Tb emission
QD705 absorption
5E6
rel. PL intensity
ε / M-1.cm-1
1,5E6
0
450
rel. PL intensity
450
500
550
600
Wavelength / nm
650
0,00
700
SI-6: Overlap between the QD absorption spectra (black) and area-normalized Tb emission spectra (green).
S4
SI-7: Photographs of Pen-functionalized QD705 (1/4), QD605 (2/5) and QD530 (3/6) taken two years after
aqueous phase transfer with Pen. 1-3 under room light, 4-6 under UV light.
Table S1: Optical characterization of QD-F(ab) conjugates were performed by excitation at 405 nm using as
excitation sources a continuous-wave Xe lamp and a pulsed diode laser from Edinburgh Instruments for steadystate and time-resolved measurements respectively.
InPZnS/ZnSe/ZnS530
CdSe/ZnS605
CdSe/ZnS705
-Pen
-F(ab)
-Pen
-F(ab)
-Pen
-F(ab)
λ emission(nm)
505±1
502±1
602±2
606±0.5
699±2.5
703±0.5
Averagelifetime(ns)
24.3±6.1
17.7±2.8
4.6±0.4
10.4±1.1
79.4±20.8 40.7±6.4
Sample
S5
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