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Methods Mol Biol. 2019 Feb 16. doi: 10.1007/7651_2019_211. [Epub ahead of print]

Step-by-Step Protocol for Superparamagnetic Nanoparticle-Based Plasma Membrane Isolation from Eukaryotic Cell.

Author information

1
EnvirotransgeneĀ® Bio-solutions Global Chennai India and Institute of Cancer Research, Oslo University Hospital, Oslo, Norway. deepak.balaji@bitsaa.org.
2
Laboratory of Lipid Metabolism and Cancer, Louvain, Belgium.
3
Bannari Amman Institute of Technology, Sathyamangalam, TN, India.
4
Faculty of Biology, Hanoi National University of Education, Hanoi, Vietnam.
5
Institute for Research and Development, Duy Tan University, Danang, Vietnam.
6
School of Odonto Stomatology, Hanoi Medical University, Hanoi, Vietnam.

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.

KEYWORDS:

Phospholipids; Plasma membrane and pulse-chase; Superparamagnetic nanoparticles

PMID:
30771189
DOI:
10.1007/7651_2019_211

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