Background of Lysosome Purification by Superparamagnetic Nanoparticles
Lysosomes are dynamic organelles that serve as regulatory centers of cellular homeostasis. In lysosomal storage diseases, lysosomes redistribute or diffuse in conventional density gradient methods resulting in reduced yield and purity. It was therefore not possible to generate enough pure material for functional lysosomal assays. Superparamagnetic iron oxide nanoparticles (SPIONs) are nanoparticles between 50 and 180 nm. It has attracted much attention due to its excellent superparamagnetic properties such as controllable size, large specific surface area, and non-toxicity. In the field of biomedicine, SPIONs are currently the most widely used functional nanomaterials due to their unique physical, chemical, magnetic and biocompatibility, such as contrast agents, iron replacement therapy, drug delivery, tissue repair, hyperthermia, cellular and tissue targeting and transfection. Researchers have successfully used SPIONs to rapidly isolate lysosomes from tissue culture cells.
Fig. 1. Superparamagnetic nanoparticles for lysosome isolation to identify spatial alterations in lysosomal protein and lipid composition. (Tharkeshwar A K, et al., 2020)
Our Superparamagnetic Nanoparticles Lysosome Purification Services
We are very enthusiastic about using nanobiotechnology methods to isolate lysosomes and study their biological relevance. For many years, CD BioSciences has focused on the synthesis and use of SPIONs, and has established a platform to synthesize SPIONs for the isolation of endosomes and lysosomes. Here, we provide a method for isolation of lysosomes using pulse-chasing superparamagnetic nanoparticles that can purify high yields of ultrapure functional lysosomes from a variety of cell types, including lysosomal storage barriers.
SPIONs target the lumen of lysosomes through the endocytic pathway and accumulate in lysosomes allowing lysosomes to be magnetically separated. Our engineers developed a step-by-step protocol for the isolation of SPIONs-based endosomes and lysosomes.
(1) We use thermal decomposition to synthesize superparamagnetic nanoparticles with iron oxide (Fe3O4) cores.
(2) We performed a ligand exchange strategy for the surface functionalization of SPIONs with dimercaptosuccinic acid (DMSA).
(3) We isolated endosomes and lysosomes from HeLa cells using the generated DMSA-SPIONs.
Why Choose Us
- Our professional team is very familiar with the principles of SPIONs and potential pitfalls can be avoided.
- A fast and robust nanoparticle-based magnetic separation method.
- Obtain high-purity, high-yield lysosomes.
- Purified lysosomes are suitable for in vitro fusion assays, lipidomic and proteomic analysis.
- Provides theoretical support for potential functional analysis of lysosomes.
CD BioSciences can meet any reasonable needs of our clients, taking time and budget into consideration for you. Our aim is to be customer-centric and to provide the highest quality services to customers. Our customer service representatives are enthusiastic and trustworthy 24 hours a day, 7 days a week. If you are interested in our services, please feel free to contact us for more information or a detailed discussion.
References
- Tharkeshwar A K, Demedts D, Annaert W. (2020) Superparamagnetic nanoparticles for lysosome isolation to identify spatial alterations in lysosomal protein and lipid composition[J]. STAR protocols. 1(3): 100122.
- Walker M W, Lloyd-Evans E. (2015) A rapid method for the preparation of ultrapure, functional lysosomes using functionalized superparamagnetic iron oxide nanoparticles[M]//Methods in cell biology. Academic Press. 126: 21-43.