Products

Exosome Isolation Tools, Immunoplates for exosome capture

To isolate overall exosomes or specific exosome sub-populations from a wide range of biofluids

or cell media

HansaBioMed offers different Immunoplates for exosome capture and/or enrichment

HBM Immunoplates are 96 multiwell plates covalently pre-coated with specific exosome-binding antibodies

enabling exosome capture and isolation from different media (cell culture supernatants, human plasma,

serum, urine and saliva). The immunocapture approach maximizes the quantity of captured exosomes,

compared to ultracentrifugation, resulting in higher exosome yields, with greater purity. HBM has

developeddifferent type of plates for the enrichment of specific exosome subpopulations (tumor, neural, glial,

monocytes and platelets) for dedicated applications. Plates are blocked and stabilized for long term storage

and ziplocked in individual alluminium bags.

Capture of overall Exosomes

Plates are coated with antibodies against exosome surface antigens, present on overell exosome population.

- From human biofluids (plasma, serum, urine, saliva)

- From cell media of human or mouse cell lines

Capture of specific Exosome sub-population

Plates are coated with antibodies overexpressed in particular pathological conditions on exosome surface.

- Capture from human plasma

- Capture and enrichment of tumor, glial, neuro, platelet derived exosomes

Applications

• Multiple profiling of exosomal markers from a single sample or screening of a larger number of samples

•  Exosome capure and quantification from human biofluids (plasma, serum, urine, saliva)

• Exosome capture and quantification from human or mouse cell media

• Titration of purified exosomes

• Capture and enrichment of specific exosome subpopulations

Advantages

• Ready to use

• Long term storage (up to 2 years)

• No exosome pre-purification required (by ultracentrifugation or other methods)

• Small amount of sample required (100 μl per well)

• Suitable for nucleic acid extraction from exosome captured on the plate

• Flexibility in designing a multiplexing assay

• Open platform for customized coating solutions

Immunoplates allow exosome protein profiling without pre-purification (by ultracentrifuge,

chemical precipitation or microfiltration).

HBM plates are useful tools for immunocapturing exosomes from biofluids or culture media, for protein

analyses and protein marker profiling. They allow quantitative and qualitative simultaneous analysis of

different protein markers (Fig 1) from the same sample, or expression profiling of a single marker in different

samples (Fig 2), without exosome pre-purification via ultracentrifuge or other methods. CD9 expression

(Fig 3) of immunocaptured plasma exosomes mimic that of the correspondent ultracentrifuged fraction. No

significant cross-reactivity is observed with soluble antigens or other vesicleassociated proteins (Fig 4).

Fig. 1. Common exosomal biomarkers analysis in a healthy donor's plasma sample

Fig. 2. CD63 profiling on exosomes derived from supernatants of different cell lines

Fig. 3. Comparison of CD9 detection on purified (ultracentrifuged) plasma exosomes vs unfractionated samples in

a set of healthy donor's plasma

Fig. 4. HBM plate is selective in capturing purified exosomes (pellet after centrifugation 120000g) and no other

circulating microvesicle (pellet 1200g and 10000g)

Immunoplates enable enrichment in tumor-derived exosome subpopulation from human

plasma

Tumor-derived Exosome capture Immunoplates are precoated with a proprietary capturing antibody which

allows enrichment and selection of exosomes from tumor origin from human plasma, particularly useful for

cancer biomarker profiling (Fig 5). Figure 6 shows that by simply using the Tumor-derived exosome plate it

is possible to discriminate healthy donors from cancer patient (indicated by a black arrows).

Fig. 5. Enrichment in Tumor-derived exosome in early and late stage melanoma (Mel E; Mel ADV) and ovary

carcinoma (Ova E, Ova ADV) using Tumor-derived exosome capture plate vs Overall exosome capture plate

(blank subtracted).

Fig. 6.  Immunoplate for Tumor-derived exosomes capture allows discrimination of cancer patients

(black arrows)

Specific Immunoplates for enrichment in neural, glial, platelet-derived exosome

subpopulation from human plasma

These plates are precoated with exosome associated antibody indicative of neurological, glial or

monocyte/platelet origin enabling specific enrichment of exosomes from human plasma samples. In the

examples reported (Fig 7) we show the specificity of binding of exosome derived from neuroblastoma (SH)

or glioblastoma (U87) cell lines when plates for neural or glial-derived exosome capture are used.

In figure 8 and 9, increasing amount of purified exosomes from neuroblastoma (SH) or glioblastoma (U87)

cell lines were spiked in human plasma from healthy donors. Selective capture of specific exosome

subpopulation was tested comparing the same plasma sample after spiking of purified plasma exosomes

(HD). In both experiments (Fig 8, 9) the signal relative to the specific exosomes subpopulation increases

after spiking, while no changes are detectable with HD, suggesting the enrichment of neural or glial derived

exosomes respectively.

Fig. 7. Specific immunocapture of 30 μg of exosome isolated from Neuroblastoma (SH-30) or Glioblastoma

(U87-30) cell lines in specific immunoplates. COLO cell and plasma (PEP) purified exosomes were used as

controls.

Fig. 8. Immunocapture of neuroblastoma derived exosomes (SH) subpopulation diluted in human plasma

from healthy donors.

Fig. 9. Immunocapture of glioblastoma derived exosomes (U87) subpopulation diluted in human plasma from

healthy donors