Trehalose SG - Exosomes brochure

Hayashibara ― A world leading supplier of pharmaceutical grade Trehalose SG

Founded as a starch syrup manufacturer in 1883, Hayashibara has grown into a company with strong research and development expertise, engaging in original and creative research. Utilizing our technological background in biotechnology and in-depth knowledge of saccharide production, Hayashibara became the first company in the world to succeed in the mass production of “Trehalose”, a unique disaccharide recognized as a beneficial excipient for the biopharmaceutical industry. In addition to trehalose, Hayashibara has been supplying for many years other high purity ingredients to the pharmaceutical industry worldwide.

Trehalose SG – Exosomes

What is exosomes?

Trehalose SG – Exosomes are membrane vesicles of about 30 nm to 200 nm in diameter secreted by most cells and are observed in vivo in body fluids such as saliva, blood, urine, and milk, and are also secreted by cultured cells.
Utilizing the properties of exosomes, research is being conducted on their use as drug delivery systems (DDS), and exosomes derived from bovine milk are attracting attention as a means of orally administering anti-cancer drugs.

Characterization of bovine milk exosomes derived by ultracentrifugation

Materials & Methods

Non-fat bovine milk

Addition of acetic acid (final conc. 1%) and mix well
Centrifugation at 1500 × g for 30 min at 4℃
Filtration using 0.22 μm filter

Whey

Ultracentrifugation at 150,000 × g for 70 min at 4℃
Pellet (exosomes) was suspended in PBS
Ultracentrifugation at 150,000 × g for 70 min at 4℃
Pellet was resuspended in PBS
Centrifugation at 10,000 × g for 5 min at 4℃

Exosome (Western blotting)

Protein concentration was measured by BCA method
Seventy μg of whey or 5 μg of bovine milk exosomes were applied on SDS-PAGE
Blotted onto PVDF membrane
Staining with anti-CD9, CD63, or CD81 antibody, respectively
(1:1,000, SBI System Biosciences)
Washing the membrane 3 times for 5 min
Staining with HRP-conjugated goat anti-rabbit antibody
(1:10,000, SBI System Biosciences)
Washing the membrane 3 times for 5 min
Detection using ECL Select™ Western Blotting Detection System (GE Healthcare).

Results

The milk exosomes derived from whey by ultracentrifugation were positive for CD9, CD63 and CD81.
As far as tested by Western blotting, respective positive band detected by anti-exosome related antibody was not found in the whey.

Hayashibara Trehalose SG Exosome_Figure 1 — Fig.1. Characterization of bovine milk exosomes by Western blotting. No.1; whey (70 μg), No. 2; bovine milk exosomes (5 μg)

Inhibitory effect of Trehalose SG on exosome aggregation by lyophilization

Hayashibara Trehalose SG Exosome_Figure 2

See the full brochure on “Hayashibara Trehalose SG – Exosomes” here

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Source: Hayashibara brochure “Hayashibara Trehalose SG – Exosomes”

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