Abstract
Here, we elaborate our detailed protocol for synthesis, functionalization, and application of superparamagnetic nanoparticle (SPMNP) for plasma membrane and lysosome isolation. We used standard thermal decomposition-based synthesis of iron oxide (Fe3O4) core SPMNP 1.0. Using ligand addition methodology, we surface functionalized SPMNP 1.0 with phospholipids and generated phospholipid-SPMNP 2.0. Further we used NH2-phospholipid-SPMNP 2.0 to isolate plasma membrane. Using our SPMNP subcellular fractionation protocol, we are able to isolate high-pure-high-yield plasma membrane using NH2-phospholipid-SPMNP 2.0. As a future perspective, we propose to use SPMNP on clinical patient samples and perform mass spectrometry-based proteomics, lipidomics, and glycomics for early cancer diagnosis.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Thimiri Govinda Raj DB et al (2011) A novel strategy for the comprehensive analysis of the biomolecular composition of isolated plasma membranes. Mol Syst Biol 7:541
Sun S, Zeng H (2002) Size-controlled synthesis of magnetite nanoparticles. J Am Chem Soc 124(28):8204–8205
Dubertret B, Skourides P, Norris DJ, Noireaux V, Brivanlou AH, Libchaber A (2002) In vivo imaging of quantum dots encapsulated in 39 phospholipid micelles. Science 298(5599):1759–1762
Raj DBTG (2011) Superparamagnetic nanoparticle based (10 nm) isolation of plasma membrane for high resolution mass spec. analysis of the proteome, lipidome and glycome. In: Semiconductor physics section. KU Leuven, Leuven
Thimiri Govinda Raj DB, Khan NA (2018) Surface functionalization dependent subcellular localization of superparamagnetic nanoparticle in plasma membrane and endosome. Nanoconvergence 5:4
Thimiri Govinda Raj DB, Khan NA (2018) Synthesis of hybrid gold nanoparticles functionalized superparamagnetic nanoparticles. Micro-Nano Lett 13(3):292. https://doi.org/10.1049/mnl.2017.0574
Thimiri Govinda Raj DB, Khan NA (2018) Protocol for eukaryotic plasma membrane isolation using superparamagnetic nanoparticles. J Magn Magn Mater 476:628–631. https://doi.org/10.1016/j.jmmm.2017.12.070
Thimiri Govinda Raj DB, Khan NA (2017) Designer nanoparticle: nano-biotechnology tool for cell biology. Nano Convergence 3(1):22
Acknowledgments
This work was supported by Envirotransgene® Global. Authors thank the infrastructural support from Bannari Amman Institute of Technology, India.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media New York
About this protocol
Cite this protocol
Thimiri Govinda Raj, D.B., Khan, N.A., Venkatachalam, S., Chu, D.T. (2019). Step-by-Step Protocol for Superparamagnetic Nanoparticle-Based Plasma Membrane Isolation from Eukaryotic Cell. In: Turksen, K. (eds) Stem Cell Nanotechnology. Methods in Molecular Biology, vol 2125. Humana, New York, NY. https://doi.org/10.1007/7651_2019_211
Download citation
DOI: https://doi.org/10.1007/7651_2019_211
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0359-8
Online ISBN: 978-1-0716-0360-4
eBook Packages: Springer Protocols