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Gentle and Effective Plasma Exosome Isolation Strategy

Authors: Morávek Ondřej | Bednářová Šárka | Škultéty Ľudovít | Kozovská Zuzana | Kupčík Rudolf | Bílková Zuzana

Year: 2024

Type of publication: ostatní - přednáška nebo poster

Page from-to: nestránkováno

Titles:

Language	Name	Abstract	Keywords
eng	Gentle and Effective Plasma Exosome Isolation Strategy	Exosomes, small extracellular vesicles with a diameter of 30-200 nm, are formed through a complex biogenetic process. These nano-sized vesicles are released by cells into the extracellular space, where they play a critical role in intercellular communication. They carry proteins, lipids, RNA, and other biomolecules, enabling them to influence recipient cells and contribute to various physiological and pathological processes, as demonstrated by numerous studies. Due to their complex functions, exosomes are also implicated in diseases such as cancer and neurodegenerative disorders. This makes exosomes – particularly those originating from tumor cells, known as oncosomes – promising candidates for investigating disease pathogenesis and facilitating early, accurate diagnosis. However, exosome research faces significant challenges, particularly in isolating them with high quality and purity. Their size, composition, and origin heterogeneity complicates standardization, often leading to selective enrichment and biased findings. Common issues include low yield, contamination with non-exosomal components, and structural damage during isolation. This variability affects reproducibility and accurate quantification, especially in samples with low exosome concentrations. To address these challenges, unlike the widely used ultracentrifugation-based techniques, researchers are increasingly utilizing advanced techniques like size-exclusion chromatography, which improves yields and reduces contamination, enhancing the quality of isolated exosomes. Standardized protocols and quality control measures, including monitoring markers such as CD63 and CD81, are being implemented to enhance isolation reliability. This study introduces a useful combination of centrifugation, filtration, and low-pressure size-exclusion chromatography, that provides robustness, high capacity, and efficient recovery of intact exosomes with minimal contamination. Techniques such as SDS-PAGE, dynamic light sc	Exosomes; chromatography; low-pressure; size-exclusion chromatography