Step-by-Step Protocol for Superparamagnetic Nanoparticle-Based Endosome and Lysosome Isolation from Eukaryotic Cell

  • Deepak B. Thimiri Govinda RajEmail author
  • Niamat Ali Khan
  • Srisaran Venkatachalam
  • Dinh Toi Chu
  • Sivakumar Arumugam
Part of the Methods in Molecular Biology book series (MIMB, volume 2125)


Here, we report our step-by-step protocol for superparamagnetic nanoparticle (SPMNP)-based endosome and lysosome isolation from HeLa. Briefly, we synthesized SPMNP 1.0 with iron oxide (Fe3O4) core using thermal decomposition method. Further, we performed ligand-exchange strategy for surface functionalization of SPMNP 1.0 with dimercaptosuccinic acid (DMSA). Thus, we generated DMSA-SPMNP 2.0 and used DMSA-SPMNP 2.0 to isolate endosomes and lysosome from HeLa cells. Using our SPMNP subcellular fractionation protocol, we are able to isolate high-pure-high-yield lysosomes using DMSA-SPMNP 2.0 for lysosome proteomics and lipidomics in order to better understand subcellular compartments.


Superparamagnetic nanoparticles DMSA Endosomes Lysosomes and pulse-chase 



This work was supported by Envirotransgene® Global. Authors thank the infrastructural support from Bannari Amman Institute of Technology, India.


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Copyright information

© Springer Science+Business Media New York 2019

Authors and Affiliations

  • Deepak B. Thimiri Govinda Raj
    • 1
    Email author
  • Niamat Ali Khan
    • 2
  • Srisaran Venkatachalam
    • 3
  • Dinh Toi Chu
    • 4
    • 5
    • 7
  • Sivakumar Arumugam
    • 6
  1. 1.Envirotransgene® Bio-solutions Global Chennai India and Institute of Cancer ResearchOslo University HospitalOsloNorway
  2. 2.Laboratory of Lipid Metabolism and CancerLouvainBelgium
  3. 3.Bannari Amman Institute of TechnologySathyamangalamIndia
  4. 4.Faculty of BiologyHanoi National University of EducationHanoiVietnam
  5. 5.Institute for Research and DevelopmentDuy Tan UniversityDanangVietnam
  6. 6.School of Bio Sciences and Technology (SBST)VIT UniversityVelloreIndia
  7. 7.School of Odonto StomatologyHanoi Medical UniversityHanoiVietnam

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