# 3030

Fangting Wu* and Travis Antes

ExoQuick

An Exosome Isolation System for Serum and Bio uid-based Cancer Biomarker Discovery
Highlights
Exosomes are released from tumors in high abundance Exosome cargo re ects the origin and physiological state of the source cells MicroRNAs are found in high abundance in circulating exosomes ExoQuick easily precipitates exosomes from patient bio- uids Higher purity exosome recovery with a simple protocol Discover novel disease-speci c biomarkers with ExoQuick

1616 N. Shoreline Blvd. Mountain View, CA 94043

Exosomes - MicroRNAs - Cancer Biomarkers

Exosomes are 40100 nm membrane vesicles secreted by most cell types, including all cancer cells. While the exact biological functions of exosomes remain unclear, exosomes can mediate communication between cells, facilitating processes such as antigen presentation and trans- signaling to neighboring cells. The abundance of exosomes present in the circulation of cancer patients increases with tumor grade. Studies identifying protein and microRNA biomarkers in these circulating exosomes can reveal the tumor type and origin. These recent studies highlight the growing use and need of standardized exosome enrichment. What is missing in this research area is a method for simple, rapid and reproducible exosome puri cation to facilitate research into the role of exosome in both physiological and pathological process, as well as possible diagnostic and therapeutic potentials. In this study, SBI utilizes proprietary polymer mixture (ExoQuick) to precipitate exosomes from human serum and tumor ascites uid samples. Exosome precipitation from serum samples was veri ed by Western blot using antibodies against two commonly found exosome biomarker proteins, CD63 and CD9. In addition, ExoQuick is capable of e ciently precipitating tumor exosomes from ovarian tumor ascites uid with greater purity, more exosomeal protein content and higher amounts of exosomal microRNAs than all other current methods. We present a new approach using ExoQuick to facilitate high-throughput exosome analysis which will enable major strides forward in circulating microRNA biomarker discoveries for cancer diagnostic and prognostic tools.

ExoQuick works with Tumor Ascites Fluid

Isolate Exosomes from Ovarian Tumor Ascites uid

Enrich for more exosomal proteins

2.0

Isolate more exosomal microRNAs

ExoQuick
[FU] 30 25 20 15 10 5 0

Exosomal Protein (mg/ml)

1.6 1.2 0.8 0.4 0.0

ExoQuick

Chromatography DynaBeads Ultracentrifugation

Small RNA: 16,660.5 pg/l microRNA: 8,904.1 pg/l

Ratio: 53%

microRNAs
25 30 35 40 45 50 55 60 65 [s]

Ascites 1

Ascites 2

Ascites 3
[FU]

Whole Ascites

Placental Alkaline Phosphatase Biomarker Enrichment

Chromat.
As1 As2

60 50 40 30 20 10 0

DynaBeads
As2

Ultracent.

ExoQuick

Small RNA: 5,297.3 pg/l microRNA: 1,119.5 pg/l Ratio: 21%

As3 As1

As3 As1 As2 As3 As1 As2 As3

PLAP

microRNAs
25 30 35 40 45 50 55 60 65 [s]

DynaBeads

ExoQuick Easily Precipitates Exosomes

Serum or Bio-fluid

While each method puri es exosomal material, circulating exosomes isolated by ExoQuick precipitation produces exosomal RNA and protein with greater purity and quantity than chromatography, ultracentrifugation, and anti-EpCAM magnetic DynaBeads. This will enhance the sensitivity and accuracy of down-stream analyses, such as qRT-PCR pro ling of microRNA and mass spectrometric and electrophoretic analyses of exosomal proteins.

[FU] 30 25 20 15 10 5 0 25

Small RNA: 11,116.0 pg/l microRNA: 5,255.4 pg/l Ratio: 47%

Simple one-step precipitation

Isolate exosomes with ease

No time-consuming ultracentrifugation Less expensive than costly DynaBeads More eective than any other method Use as little as 250l of serum or bio- uid

Exosome marker proteins analysis

(26 kDa) kDa MW
55 35 27

microRNAs
30 35 40 45 50 55 60 65 [s]

CD63

(25 kDa)

CD9

Supernatant
ExoQuick + Solution

Sup EXO Sup EXO

Use ExoQuick to isolate exosomes from:

Combine ExoQuick with SBIs microRNA miRNome qPCR arrays to rapidly nd microRNA signatures. Patient Serum and Plasma Patient Urine
1000.00 100.00

Patient Ascites uid Works even with tissue culture media

1 2 3 4 5 6 7 8 9 10 11 12 A B C E D F H G

Exosome pellet

10.00

15

1.00

0.10

0.01

0.00

Western blot analysis

www.systembio.com