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RSS FeedsIJMS, Vol. 22, Pages 11144: Human Serum Extracellular Vesicle Proteomic Profile Depends on the Enrichment Method Employed (International Journal of Molecular Sciences)

 
 

15 october 2021 15:20:37

 
IJMS, Vol. 22, Pages 11144: Human Serum Extracellular Vesicle Proteomic Profile Depends on the Enrichment Method Employed (International Journal of Molecular Sciences)
 


The proteomic profiling of serum samples supposes a challenge due to the large abundance of a few blood proteins in comparison with other circulating proteins coming from different tissues and cells. Although the sensitivity of protein detection has increased enormously in the last years, specific strategies are still required to enrich less abundant proteins and get rid of abundant proteins such as albumin, lipoproteins, and immunoglobulins. One of the alternatives that has become more promising is to characterize circulating extracellular vesicles from serum samples that have great interest in biomedicine. In the present work, we enriched the extracellular vesicles fraction from human serum by applying different techniques, including ultracentrifugation, size-exclusion chromatography, and two commercial precipitation methods based on different mechanisms of action. To improve the performance and efficacy of the techniques to promote purity of the preparations, we have employed a small volume of serum samples (<100 mL). The comparative proteomic profiling of the enriched preparations shows that ultracentrifugation procedure yielded a larger and completely different set of proteins than other techniques, including mitochondrial and ribosome related proteins. The results showed that size exclusion chromatography carries over lipoprotein associated proteins, while a polymer-based precipitation kit has more affinity for proteins associated with granules of platelets. The precipitation kit that targets glycosylation molecules enriches differentially protein harboring glycosylation sites, including immunoglobulins and proteins of the membrane attack complex.


 
150 viewsCategory: Biochemistry, Biophysics, Molecular Biology
 
IJMS, Vol. 22, Pages 11145: Trimethylamine n-Oxide (TMAO) Modulates the Expression of Cardiovascular Disease-Related microRNAs and Their Targets (International Journal of Molecular Sciences)
IJMS, Vol. 22, Pages 11146: Synergistic Action of Antimicrobial Lung Proteins against Klebsiella pneumoniae (International Journal of Molecular Sciences)
 
 
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