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Preparation and functionalization of graphene nanocomposites for biomedical applications

Abstract

Functionalized nano-graphene– and graphene-based nanocomposites have gained tremendous attention in the area of biomedicine in recent years owing to their biocompatibility, the ease with which they can be functionalized and their properties such as thermal and electrical conductivity. Potential applications for functionalized nanoparticles range from drug delivery and multimodal imaging to exploitation of the electrical properties of graphene toward the preparation of biosensing devices. This protocol covers the preparation, functionalization and bioconjugation of various graphene derivatives and nanocomposites. Starting from graphite, the preparations of graphene oxide (GO), reduced GO (RGO) and magnetic GO–based nanocomposite, as well as how to functionalize them with biocompatible polymers such as polyethylene glycol (PEG), are described in detail. We also provide procedures for 125I radiolabeling of PEGylated GO and the preparation of GO-based gene carriers; other bioconjugation approaches including drug loading, antibody conjugation and fluorescent labeling are similar to those described previously and used for bioconjugation of PEGylated carbon nanotubes. We hope this article will help researchers in this field to fabricate graphene-based bioconjugates with high reproducibility for various applications in biomedicine. The sample preparation procedures take various times ranging from 1 to 2 d.

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Figure 1
Figure 2: A schematic to show further bioconjugation of PEGylated nano-GO (Step 3).
Figure 3: Characterization of various functionalized GO derivatives.
Figure 4: 125I-labeled nGO-PEG for the in vivo biodistribution study.
Figure 5: Graphene-based gene transfection.

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Acknowledgements

This work was supported by the National Basic Research Program of China (973 Program, 2012CB932600 and 2011CB911002), the National Science Foundation of China (NSFC, 51002100, 51222203, 51132006) and a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD), the University of Wisconsin at Madison, the US National Institute of Biomedical Imaging and Bioengineering/National Cancer Institute (1R01CA169365), and the US Department of Defense (W81XWH-11-1-0644).

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Authors

Contributions

Z.L. and W.C. designed the experiments and wrote the manuscript; K.Y., L.F. and H.H. performed the experiments, analyzed the results and wrote the manuscript.

Corresponding author

Correspondence to Zhuang Liu.

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The authors declare no competing financial interests.

Integrated supplementary information

Supplementary Figure 1 Biodistribution of free 125I and 125I-nGO-PEG at 6 h post i.v. injection.

Minimal uptake of free 125I was observed in the liver, spleen, as well as most other organs except thyroid and stomach due to the fast renal excretion of small iodine ions33.

Supplementary information

Supplementary Figure 1

(PDF 147 kb)

Supplementary Table 1

Summary of in vivo toxicity of different polymer-functionalized GO. (PDF 169 kb)

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Yang, K., Feng, L., Hong, H. et al. Preparation and functionalization of graphene nanocomposites for biomedical applications. Nat Protoc 8, 2392–2403 (2013). https://doi.org/10.1038/nprot.2013.146

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