Custom Exosome Services

Exosomes are small (~40‐150 nm) extracellular vesicles (EVs) released from all cell types and found in body fluids and cell culture supernatants. Exosomes are generated by fusion of a specialized endosome, the multivesicular body (MVB), with the plasma membrane. Exosomes have been proposed to provide means for intercellular exchange of macromolecules, allowing the transfer cellular information, contributing to intercellular communication in relevant biological processes.

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Exosome Isolation and Purification

Immunostep has developed a series of protocols for the isolation and purification of high-quality exosomes derived from multiple sources including cell culture samples and all human proximal fluids, including plasma, urine, cerebrospinal fluid, amniotic fluid, bronchoalveolar lavage fluid, synovial fluid, sperm, saliva, malignant and pleural effusions of ascites and breast milk.

Ultracentrifugation

Based on density and size under centrifugal force is generally considered being the gold standard method for extracellular vesicles isolation, although it may produce some aggregation of EVs.

Precipitation

Based on EVs solubility, is a quick method, with  high recovery although it usually precipitates non-EV material.

Size Exclusion Chromatography (SEC)

This is probably the most suitable method, alone or combined, to isolate exosomes from biological samples and in particular  from plasma/serum, since it allows the separation of exosomes from important contaminants such as proteins and HDL. Additionally, Immunostep has columns that allow separation between EVs-Exosomes and other Extracellular Vesicles.

Immunoaffinity capture

Based on the use of capture beads, it allows the specific isolation of exosomes based on the membrane display of specific marker. For example: CD63, EpCAM, PD-L1, etc.

Protocols for Exosome Purification

At Immunostep we have developed a wide range of protocols for exosome isolation/purification from all kind of samples, providing a customized solution for each project, with high purity and optimal recovery results and compatible with any downstream processing or analysis. 

Article: Most commonly used Exosome Isolation Methods

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Characterization Methods

We have developed and implemented several characterization and validation methods, for both research and clinical purposes, that allow analyze purity and composition and to quantify exosomal cargo, which contains critical information that has proved to be tremendously promising  for diagnostic, prognostic and therapeutic use in a variety of disease areas. These methods include:

Graphic representation: Exosome Composition

  • Transmission electron microscopy (TEM)

  • Scanning electron microscopy (SEM)

  • Nanoparticle tracking analysis (NTA) 

  • Western blot (WB)

  • Flow cytometry

  • Fluorescence-activated cell-sorting (FAQS)

  • Multiplexing Bead Arrays

  • Enzyme-linked immunosorbent assay (ELISA)

  • Gene expression analysis (mRNA and microRNA)

Electronic microscopy: exosomes isolated by a capture bead

Fluorescence: activated cell sorting (FACS)

Nanoparticle tracking analysis (NTA).Size and particle concentration analysis report.

Exosome Characterization

Exosome characterization and analysis presents a tremendous clinical opportunity and specifically in the area of liquid biopsy. However, realizing the potential requires detecting and characterizing exosomes with high accuracy and reproducibility, which can be challenging for any project or research. Immunostep makes available to its clients all their experience and technical capabilities in the field of exosomes, to make the clinical use of these EVs a reality.

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