Efficient removal of biological contaminants from microbially reduced graphene oxide via a TRIzol-based protocol.
6 affiliations
- 1Istituto Dei Sistemi Complessi, ISC-CNR Via Dei Taurini 19 00185 RM Rome Italy [email protected].
- 2Dipartimento di Neuroscienze, Università Cattolica Del Sacro Cuore L. go Francesco Vito, 1 00168 Rome RM Italy.
- 3Dipartimento di Chimica, Università di Roma La Sapienza P.le Aldo Moro 5 Roma 00185 Italy.
- 4Department for Sustainability, ENEA, Casaccia Research Centre 00123 Rome Italy.
- 5Fondazione Policlinico Universitario "A. Gemelli" IRCCS L. go Agostino Gemelli 8 00136 Rome RM Italy [email protected] [email protected].
- 6Istituto di Scienze e Tecnologie Chimiche "Giulio Natta", SCITEC-CNR, C/O Istituto Biochimica e Biochimica Clinica, Università Cattolica Del Sacro Cuore L. Go Vito, 1 00168 Rome RM Italy.
Abstract
Graphene-based nanomaterials, particularly graphene oxide (GO) and reduced graphene oxide (rGO), have emerged as promising platforms for biomedical applications due to their biocompatibility, high surface area, and versatile functionalization. Among methods to obtain rGO from GO the bacteria-mediated reduction occurs by the enzymatic transfer of electrons from bacterial respiratory chains to GO, leading to the partial depletion of the oxygen-based functionalities and restoration of the graphene lattice. However, rGO obtained via microbial reduction often retains bacterial contaminants, which can strongly adsorb to the rGO surface and interfere with in vitro cellular assays and optoelectronic applications. Furthermore, the large-scale production of high-quality rGO requires the effective elimination of these biological impurities. Here, we report a new protocol for the efficient purification of microbially produced rGO using a purification method based on TRIzol, a phenol-guanidinium reagent widely used for nucleic acid and protein extraction. Our protocol effectively disrupts cellular components and dissociates macromolecular complexes, enabling the removal of surface-bound contaminants. Purified rGO was characterized by physicochemical methods and evaluated in vitro. The protocol removes bacterial contaminants without detectable alteration of the rGO structural features probed by Raman spectroscopy, XPS and FE-SEM, and the purified material showed no detectable cytotoxicity toward BJ5ta fibroblasts at 100 µg mL⁻¹ after 24 and 48 h. This approach provides a practical strategy to improve reproducibility in studies involving microbially reduced graphene-based nanomaterials. The practical constraints of the method, namely reagent hazards, possible phenol carryover and reagent cost, are critically discussed, positioning it as a laboratory-scale purification step for applications in which material purity is critical.
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BibTeX
@article{palmieri2026efficient,
title = {{Efficient removal of biological contaminants from microbially reduced graphene oxide via a TRIzol-based protocol.}},
author = {Valentina Palmieri and Francesco Amato and Giulia Artemi and Maria Antonietta Grosso and Valeria Ferrara and Caterina Perfili and Ginevra Friggeri and Giordano Perini and Antonio Minopoli and Francesca Sciandra and Andrea Giacomo Marrani and Marco De Spirito and Massimiliano Papi},
journal = {RSC advances},
year = {2026},
month = oct,
doi = {10.1039/d6ra08861a},
url = {https://europepmc.org/article/MED/42948059},
note = {CC BY-NC},
}
RIS
TY - JOUR
TI - Efficient removal of biological contaminants from microbially reduced graphene oxide via a TRIzol-based protocol.
AU - Valentina Palmieri
AU - Francesco Amato
AU - Giulia Artemi
AU - Maria Antonietta Grosso
AU - Valeria Ferrara
AU - Caterina Perfili
AU - Ginevra Friggeri
AU - Giordano Perini
AU - Antonio Minopoli
AU - Francesca Sciandra
AU - Andrea Giacomo Marrani
AU - Marco De Spirito
AU - Massimiliano Papi
PY - 2026
DA - 2026/10/27
JO - RSC advances
DO - 10.1039/d6ra08861a
UR - https://europepmc.org/article/MED/42948059
AB - Graphene-based nanomaterials, particularly graphene oxide (GO) and reduced graphene oxide (rGO), have emerged as promising platforms for biomedical applications due to their biocompatibility, high surface area, and versatile functionalization. Among methods to obtain rGO from GO the bacteria-mediated reduction occurs by the enzymatic transfer of electrons from bacterial respiratory chains to GO, leading to the partial depletion of the oxygen-based functionalities and restoration of the graphene lattice. However, rGO obtained via microbial reduction often retains bacterial contaminants, which can strongly adsorb to the rGO surface and interfere with in vitro cellular assays and optoelectronic applications. Furthermore, the large-scale production of high-quality rGO requires the effective elimination of these biological impurities. Here, we report a new protocol for the efficient purification of microbially produced rGO using a purification method based on TRIzol, a phenol-guanidinium reagent widely used for nucleic acid and protein extraction. Our protocol effectively disrupts cellular components and dissociates macromolecular complexes, enabling the removal of surface-bound contaminants. Purified rGO was characterized by physicochemical methods and evaluated in vitro. The protocol removes bacterial contaminants without detectable alteration of the rGO structural features probed by Raman spectroscopy, XPS and FE-SEM, and the purified material showed no detectable cytotoxicity toward BJ5ta fibroblasts at 100 µg mL⁻¹ after 24 and 48 h. This approach provides a practical strategy to improve reproducibility in studies involving microbially reduced graphene-based nanomaterials. The practical constraints of the method, namely reagent hazards, possible phenol carryover and reagent cost, are critically discussed, positioning it as a laboratory-scale purification step for applications in which material purity is critical.
SN - 2046-2069
ER -