Table 3 Comparison of existing technologies for endosomes and lysosome isolation
From: Designer nanoparticle: nanobiotechnology tool for cell biology
Isolation methodologies | Advantages | Disadvantages |
|---|---|---|
Density gradient centrifugation; | Conventional method that can be used to isolate endosomes and lysosomes along other subcellular compartments Effective method to isolate lysosomes from tissue or in vivo cell fractions Simple procedure that can be performed using an ultracentrifuge | Low yield Difficulties in separating endosomes from lysosome vesicles Difficulties in separating different endosomal and its associated vesicles |
Antibody based pull-down assay | Can be used to pull down proteins after post-fractionation Can also be used to target selective endocytic uptakes Can be used in combination with biotin-streptavidin assay | Limited applicability for certain endocytosis uptake Limited yield and low purity Cannot isolate vesicles under native conditions |
SPMNPs based isolation; | A novel strategy that is generic for any kind of cell systems with reasonable purity and yield Method does not involve the use of detergent or antibody that affects native conditions Method can also use targeted endosomal uptake pathway by using ligand tagged nanoparticle Can be used to isolate protein under native conditions and all endosomal uptake | Technology has not be established for isolation of vesicles form tissue cells and in vivo experiments Technology has not been established to isolate vesicles from cell suspension. Possibility exists to use this technology for cells in suspension culture |