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Extracell Vesicles Circ Nucleic Acids 2020;1:20-56 I http://dx.doi.org/10.20517/evcna.2020.10 Page 41

27. Use of high-capacity membranes for simple, rapid exosome isolations with high yield and
purity

Authors: Yi Zhao*, Boris Levitan, Brenda Huang, Micheal Haugwitz, Andrew Farmer
E-mail: yi_zhao@takarabio.com
Affiliations: Takara Bio USA, Inc, Mountain View, CA, USA.
Abstracts:
Introduction: Despite their small size, exosomes play important roles in normal physiological processes
(e.g., immune response, neuronal function, and stem cell maintenance) and diseases (e.g., cancer and liver
disease). The isolation of the vesicles has historically been accomplished via ultracentrifugation. However,
ultracentrifugation is time-consuming, is not scalable, requires specialized equipment, may damage vesicles
during the high-speed spins, and suffers from low yield. More recently, precipitation solutions have been
utilized to simplify exosome isolation protocols, but precipitation-based techniques are often inconsistent,
with low yield and reduced purity. Thus, there is a significant need for a method to isolate exosomes
without compromising purity or yield rapidly.
Methods: Here we describe the use of novel membranes conjugated to a proprietary, non- antibody based
exosome-binding compound to isolate exosomes selectively. The membranes, which we have named
Capturem™ membranes, have been chemically modified to have increased surface area, which allows higher
binding capacity while providing highly pure and concentrated samples. Additionally, the membranes have
been assembled into benchtop centrifuge-compatible spin columns, which can be used to isolate exosomes
from plasma in under 30 minutes.
Results and Conclusions: Isolations performed with the Capturem exosome isolation spin columns
produced exosomes of sizes comparable to experimental values reported in the literature; containing the
key exosome protein markers CD63, CD9, and Alix; and with little or no expression of the exosome-
negative markers Calnexin and Albumin. As a whole, the Capturem columns enable researchers to study
exosomes to accelerate the pace of their research by obtaining high yields of noncontaminated exosomes
simply and rapidly.

28. Detection of EGFR mutations in extracellular vesicle RNA and protein corresponds to
disease status in metastatic lung cancer patients

1,2
1,2#
1,2#
1,2
1,2
Authors: Emma Purcell , Sarah Owen , Emily Prantzalos , Abigail Radomski , Mina Zeinali ,
3,4*
1,2
1,2
Nayri Carman , Nithya Ramnath , Sunitha Nagrath *
E-mail: eapurcel@umich.edu
Affiliations:
1 Department of Chemical Engineering, University of Michigan, Ann Arbor, MI, USA.
2 Biointerface Institute, University of Michigan, Ann Arbor, MI, USA.
3 Department of Internal Medicine, University of Michigan Rogel Cancer Center, Ann Arbor, MI, USA.
4 Veterans Administration, Ann Arbor Healthcare System, Ann Arbor, MI, USA.
Abstracts:
Introduction: Current cancer detection and characterization methods require a lung tissue biopsy, an
invasive procedure, performed only when the patient is showing symptoms that often only arise in late-
stage cancers. Extracellular vesicles offer a stable, abundant biomarker in the blood to serially profile
molecular characteristics of patient tumors through a non-invasive liquid biopsy. In non-small cell lung
cancer (NSCLC), identifying the presence of sensitizing and resistance epidermal growth factor receptor
(EGFR) mutations informs sensitivity to tyrosine kinase inhibitors (TKIs) and dictates treatment regimes.