Particle-containing lysosomes can be enriched from a variety of cell types using superparamagnetic iron oxide nanoparticles (SPIONs). This approach has been widely used in comparative proteomic experiments of lysosomes as well as in lipidomic studies. Here, CD BioSciences provides an efficient service for the identification of lysosomal proteins by differential SPIONs treatment to support lysosomal proteomic studies.
Background of lysosomal Proteins by Differential SPIONs Treatment Identification
Currently, scientists have developed various methods to purify lysosomes. Among them, magnetic nanoparticles (ferrodextran particles (FeDex)/SPIONs) were delivered to the lysosomal compartment by nonspecific liquid-phase endocytosis, where SPIONs accumulated and gradually degraded in lysosomes. Then, lysosomes can be isolated by a magnetic field. Compared to other strategies, SPIONs' approach achieves an increase in identified lysosomal proteins and higher reproducibility of protein intensities for label-free quantification. Scientists have successfully studied the lysosomal proteome of mouse embryonic fibroblasts by SPIONs and LC-MS/MS, identifying 47,000 proteins of lysosomal-enriched fractions.
Fig. 1. Superparamagnetic iron oxide nanoparticles induced ferritin degradation by lysosomal cathepsins. (Laskar A, et al., 2012)
Our lysosomal Proteins by Differential SPIONs Treatment Identification Services
We offer an efficient service for the purification of lysosomes from superparamagnetic nanoparticles. However, we face the challenge that lysosomal sample preparation varies greatly between different data sets and there is no perfect solution for lysosomal histological analysis. Here, CD BioSciences is dedicated to perform LC-MS/MS analysis of lysosomal enriched fractions to identify a large number of lysosomal proteins.
Our engineers have successfully developed an optimized SPIONs purified lysosomal proteomic analysis protocol to identify the true lysosomal proteins. The protocol is based on SPIONs, SILAC (stable isotope labeling of amino acids in cell culture) and a bimodal distribution model for data analysis. In order to provide satisfactory lysosome identification services to our customers worldwide, we take the following optimization measures.
- Systematic evaluation and optimization of the main steps of lysosome-enriched fraction preparation.
- Mass spectrometry analysis of the lysosome-enriched fraction.
- Generation of a library of spectra for analysis of lysosomal proteins by data independent acquisition (DIA).
- Analysis of lysosomal proteins in combination with solid phase extraction resins, peptide separation and concentration methods.
Our Advantages
- Advanced technical support such as LC-MS/MS, solid-phase extraction resins, peptide separation and concentration methods.
- Enrichment of lysosomal proteins is not required, samples are lysosomal enriched fractions.
- Potential new lysosomal proteins can be identified.
- Enables the identification of selected candidate lysosomal localizations.
- Performs correlation analysis on individual cell lines and calculates how often each protein is identified in the putative lysosomal fraction.
- Allows for the assignment of potential new lysosomal proteins.
We look forward to collaborating with you. You are always welcome to engage in discussions with us at any point of the project. If you are interested in our services, please feel free to contact us for more information.
References
- Laskar A, Ghosh M, Khattak S I, et al. (2012) Degradation of superparamagnetic iron oxide nanoparticle-induced ferritin by lysosomal cathepsins and related immune response[J]. Nanomedicine. 7(5): 705-717.
- Ponnaiyan S, Akter F, Singh J, Winter D. (2020) Comprehensive draft of the mouse embryonic fibroblast lysosomal proteome by mass spectrometry based proteomics. Sci Data. 7(1):68.