MilliporeSigma
  • Home
  • Search Results
  • Isolation and mutational assessment of pancreatic cancer extracellular vesicles using a microfluidic platform.

Isolation and mutational assessment of pancreatic cancer extracellular vesicles using a microfluidic platform.

Biomedical microdevices (2020-03-13)
Nabiollah Kamyabi, Reza Abbasgholizadeh, Anirban Maitra, Arezoo Ardekani, Sibani L Biswal, K Jane Grande-Allen
ABSTRACT

Cancer cells release extracellular vesicles known as extracellular vesicles (EVs), containing tumor-derived DNA, RNA and proteins within their cargo, into the circulation. Circulating tumor-derived extracellular vesicles (TEV) can be used in the context of serial "liquid biopsies" for early detection of cancer, for monitoring disease burden in patients, and for assessing recurrence in the post-resection setting. Nonetheless, isolating sufficient TEV by ultracentrifugation-based approaches, in order to enable molecular assessment of EVs cargo, can be an arduous, time-consuming process and is inconsistent in the context of yield and purity among institutions. Herein, we describe a microfluidic platform, which we have named MITEV (Microfluidic Isolation of Tumor-derived Extracellular Vesicles) for the rapid isolation of TEV from the plasma of pancreatic cancer patients. The device, which has ~100,000 pillars placed in a zigzag pattern and is coated with antibodies against generic EV surface proteins (anti-CD63, -CD9, and -CD81 antibodies) or the TEV specific anti-Epithelial Cell Adhesion Molecule (EpCAM) antibody, is capable of high-throughput EVs isolation and yields sufficient DNA (total of ~2-14 ng from 2-ml plasma) for downstream genomic analysis. Using two independent quantitative platforms, droplet digital polymerase chain reaction (ddPCR) and molecular barcoding using nanoString nCounter® technology, we can reliably identify KRAS mutations within isolated TEV of treatment-naïve metastatic pancreatic cancer patients. Our study suggests that the MITEV device can be used for point-of-care applications, such as in the context of monitoring residual or recurrent tumor presence in pancreatic cancer patients undergoing therapy.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
(3-Mercaptopropyl)trimethoxysilane, 95%
Sigma-Aldrich
4-Maleimidobutyric acid N-hydroxysuccinimide ester, ≥98.0% (HPLC)