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Supplementary material to the article available at:
https://doi.org/10.5802/crchim.251
Supplementary material: 1.4 nm gold
nanoparticle-antibody conjugates for in situ gold
immunolabelling after transduction into living
human cells
Nadja Groysbeck a, AnneMarie Haeberléb, Stéphane Ory b, Victor Hanss c,
Mikhael Eltsov c, Patrick Schultz c and Guy Zuber ∗, a
a Université de Strasbourg - CNRS, UMR7242 Biotechnologie et Signalisation
Cellulaire, 300 Bd Sébastien Brant, CS 10413, 67412 Illkirch, France
b Centre National de la Recherche Scientifique, Université de Strasbourg, Institut des
Neurosciences Cellulaires et Intégratives, F-67000 Strasbourg, France
c Centre for Integrative Biology (CBI), Department of Integrated Structural Biology,
Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), 1 rue Laurent
Fries, BP10142, F-67404 Illkirch Cedex, France
E-mails: nadja.groysbeck@gmail.com (N. Groysbeck), haeberle@inci-cnrs.unistra.fr
(A. M. Haeberlé), ory@inci-cnrs.unistra.fr (S. Ory), hanssv@igbmc.fr (V. Hanss),
eltsovm@igbmc.fr (M. Eltsov), pat@igbmc.fr (P. Schultz), zuber@unistra.fr (G. Zuber)
Not yet published
∗Corresponding author.
ISSN (electronic) : 1878-1543 https://comptes-rendus.academie-sciences.fr/chimie/
2 Nadja Groysbeck et al.
Supplementary Figure S1. Controls experiments. Image of 4% PFA-fixed and 0.1% Triton X-100 perme-
abilized HeLa cells. (A) After permeabilization, the cells were treated with silver-enhancement solution.
(B) Gold immunolabelling of HeLa with the 7G5AuPEG anti-RPB1 without silver-enhancement solution.
Nadja Groysbeck et al. 3
Supplementary Figure S2. Analysis of the production of 7G5AuC and binding specificity using fixed
HeLa cells. (A) The 7G5 was firstly reduced with TCEP, reacted with Auz and purified by size-exclusion
chromatography. The purified conjugate was then reacted for passivation with the peptide CALNNG (C).
(B) The CALNNG appeared to promote unspecific binding of the probe the fixed cell ultrastructure.
4 Nadja Groysbeck et al.
Supplementary Figure S3. Immunofluorescence imaging. Otherwise indicated, HeLa cells adhered onto
glass coverslips were fixed with 4% PFA for 20 min. The plasma membrane was permeabilized with 0.1%
Triton X-100 before incubation with the indicated primary mouse mAbs in PBS containing 10% FBS. The
mouse antibody was detected with a goat anti-mouse AlexaFluor488-antibody conjugate.
Nadja Groysbeck et al. 5
Supplementary Figure S4. Non-reducing SDS-PAGE analysis of the indicated AuNP-mAb conjugates.
The proteins were stained with Coomassie blue.
6 Nadja Groysbeck et al.
Supplementary Figure S5. HeLa cells stably expressing H2B-GFP were fixed with 4% PFA for 20 min,
permeabilized with 0.1% Triton X-100. The H2BGFP fusion protein was then detected by binding of
2A3Au and 2A3AuPEG conjugates to the GFP. Detection was done either directly using the GFP green
fluorescence (top images), an anti-mouse AlexaFluor-594-Ab conjugate (middle images) or the AuNP via
a silver enhancement procedure. Results showed that coating of the AuNP with PEG limit unspecific
binding.
Nadja Groysbeck et al. 7
Supplementary Figure S6. HeLa cells were incubated with the indicated AuNP-7G5 conjugates and
treated with three pulses of 10 ms at 517 V·cm−1. After 20 h or 44 h of cell culture, the cells were fixed
with 4% PFA for 20 min, permeabilized with Triton X-100 and the gold particles were enhanced with silver.


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Electrooxidation treatment of simulated wastewater using mixed-metal oxide anodes for bacterial decontamination

Kinetic aspects of malachite deposition on marble from copper lactate solutions

Le foisonnement éolien : les limites d’un mix électrique à forte proportion d’énergies renouvelables intermittentes

Valorisation of plastic waste from the beverage industry through its transformation into adsorbent and solid fuel materials

A review on the chemical ecology of the Fucaceae holobionts: from fundamental knowledge to applications

Studying chemical signals in plant–soil mesofauna interactions

1.4 nm gold nanoparticle-antibody conjugates for in situ gold immunolabelling after transduction into living human cells

Structural characterization of stem cell factors Oct4, Sox2, Nanog and Esrrb disordered domains, and a method to detect phospho-dependent binding partners

Aquathermolysis of heavy oil catalyzed by transition metal salts and clay

Quantitative analysis of the effect of microtubule-targeting drugs on the microtubule cytoskeleton of breast cancer cells with different invasive properties

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