In addition to conventional exosomes of animal origin and exosome-like vesicles of plant origin, bacteria, fungi, and protozoa have been found to produce vesicles of similar size to exosomes. Moreover, these microorganism-derived exosomes can be used for drug delivery, study of infection mechanisms, cancer therapy, modulation of host immune response, and engineering as vaccines, etc. Creative Biolabs provides customized isolation and development services for microorganism-derived exosome research.
1. Microorganism culture
Select the appropriate medium and conditions to culture the target microorganisms. Microorganisms can be taken from bacteria, fungi, and protozoa.
2. Microorganism removal
Centrifuge the culture in a centrifuge tube to remove microorganism residue. Normally low-speed centrifugation is chosen to minimize the loss of exosomes.
3. Exosome extraction
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Collect the supernatant from which the microorganisms have been removed and centrifuge it at high speed to precipitate the exosomes. Typically, exosomes can be purified by multiple centrifugation and washing steps.
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Capturing various membrane vesicles in the bacterial culture medium based on affinity resin chromatography provides high yield and purity while removing small molecules and ions from the bacterial culture supernatant, providing an alternative to ultracentrifugation.
4. Screening and concentration
Screen the purified exosomes using filter membranes of appropriate pore size. Then, concentrate the exosomes using ultrafiltration or concentration devices.
5. Identification and characterization
The exosomes are examined and characterized using TEM and NTA. In addition, Western blot, mass spectrometry, and other techniques can be used to further characterize the protein or nucleic acid components of exosomes.
Fig. 1 Bottom-loaded density gradient ultracentrifugation for isolation of bacterial EVs.1
Fig. 2 Host- and microbial-derived extracellular vesicles in humans.2
