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냉장보관이 가능한 Lyophilzed Exosome Standards 

Exosomes are small endosomederived lipid nanoparticles (30-100nm) actively secreted by exocytosis

by most living cells.

Exosomes have been isolated from many different cell lines (hematopoietic cells, tumor lines, primary

cultures, virus infected cells) as well as from biological fluids, in particular blood (serum and plasma) and

other body fluids (pleural effusions, synovial fluid, urine, amniotic fluid, semen, saliva, bronchoalveolar

lavage fluid etc). Exosome release occurs either constitutively or upon induction, under bothnormal and

pathological conditions, in a dynamic, regulated and functionally relevant manner so that both amount  and

molecular composition of released exosomes depend on the state of a parent cell. This is one of major

premises for employing exosomes for diagnostic purposes.

HBM(HansaBioMed) offers exosomes purified from several cell lines and human body fluids to facilitate

research in the extracellular vesicle field.

HBM Lyophilized Exosome Standards may be used as standard reference in experiments for profiling of

exosome-derived proteins and nucleic acids. Finally they can be used as calibration standards for

quantitative comparison of different biological samples. 

HBM Lyophilized Exosome Standards include,

1. Exosome Standard from Human Body Fluids

   Purified exosomes from human Plasma, Serum and Urine (pools of healthy donors)

2. Exosome Standard from Cell Culture Supernatants

   Purified exosomes from supernatants of a wide list of human tumor cell lines

3. Cell lysates

   Whole cell lysates from a wide list of human tumor cell lines 

Lyophilized Exosome Standards

Lyophilization is the ideal technique for preserving the long-term stability of exosomes at 4℃.

Lyophilized exosomes can be used as control standards for multiple applications including FACS, WB,

ELISA and as calibration standards for quantitation of exosome-derived markers from biological samples.

Lyophilized exosomes are easy to ship and store, and are stable for 12 months at 4℃. Purified and

lyophilized exosomes are obtained from a variety of biological sources (cell culture supernatant, human

plasma, serum, urine and saliva). Exosomes are purified following a combination of ultracentrifugation and

microfiltration steps. Exosomes are subsequently quantified and validated for overall protein contentand

particle number by NTA (Nanoparticles Tracking Analysis) with NanoSight LM10. Lyophilization does not

affect the stability of purified exosomes and the expression levels of their exosome markers (proteins and

nucleic acids).

Applications

• Assay calibration

• Control (spike-in) for exosome quantification

• Protein marker analysis with different techinques

• Exosomal nucleic acid extraction and analysis

• Positive control for researchers in the exosome field 

Advantages

• Higher purity and better performance than competitor

• Easy to reconstitute

• Long term storage (36 months in lyophilized form, stored at 4℃)

• Available from different human tumor cell lines

• Available from human healthy donors body fluids (Plasma, Serum, Urine & Saliva) 

Lyophilization is the ideal method for preserving exosome stability

Expression of exosomal markers was assayed with different techniques (WB, FACS, ELISA) and

nanovesicle count was measured with NTA. Lyophilization did not substantially affect exosome count or

biomarker expression compared to other storage methods (Fig 1, 2, 3, 4).

Fig. 1. Western Blot comparison of exosomal markers on fresh (F),

          frozen (-20℃) and lyophilized exosomes (L) 

Fig. 2. Titration of exosome standards purified from cell culture supernatant with CD63 detection. 

Fig. 3. Comparison of exosomal markers on fresh and lyophilized exosomes'

Fig. 4. Comparative Nanosight analysis of freshly purified (right panel) and lyophilized plasma exosomes

          (left panel). 

Comparing different storage methods of exosome standards (fresh vs. frozen vs lyophilized) with an anti-

CD81 ELISA assay, the loss of signal compared to fresh material is minimal when using lyophilized

exosomes (CV - 15%) (Fig 5). 

Fig. 5. ExoTEST™ comparative detection of CD81 on HMB-MM1 derived exosomes:

         #1. Fresh exosomes

         #2. Frozen exosomes (-20℃)

         #3. Lyophilized exosomes

HBM Exosome Standards guarantee higher purity and performances over competitors

HBM Exosome Standard from human serum (HBM-PES) were compared to human serum exosome

standard from a Competitor.

Fig. 6. ELISA quantification of HBM Exosome Standard vs Competitor’s standard (human serum) for

         exosomal markers CD9 and CD63

Fig. 7. Standard curve for exosome quantification. HBM Exosome Standard vs. Competitor’s standard 

Exosomal markers CD9 and CD63 from 50 μg of both serum exosome standards were detected using an

ELISA assay (fig 6). HBM Standard (50μg) generated the highest signal, whereas the signal from Competitor

Standard (50 μg) was lower than 30 μg of HBM Standard. In a second test, 50 μg of HBM and competitor’s

Standards were serially diluted to design a standard curve for exosome quantification (fig 7). The standard

curve generated with HBM Standard was linear (R2 = 0,9876) within the concentration range and

suitable for exosome quantification.