Development of a High Throughput Exosome Isolation Microfluidic Device for Disease Diagnostics (2016)

Undergraduate: Andrianna Anderson

Faculty Advisor: Steven Soper
Department: Chemistry

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Exosomes, small vesicles (30-100 nm diameter) that are shed from cells, have cargo that contain nucleic acids indicative of their origin. It is of clinical interest to isolate exosomes for analysis of its cargo. Exosomes, like circulating tumor cells and cell-free DNA, can be isolated from patient blood, forming the basis of a liquid biopsy for the patient¿¿¿s disease state. There have been some recent advances proposing microfluidic devices to isolate exosomes, but are limited by their low-throughput, which limits the quantity of disease specific exosomes that can be isolated. We designed a microfluidic device for positive affinity selection of exosomes that can rapidly (<30 minutes) process 1 mL plasma. This device is manufactured via hot embossing of a cyclic olefin copolymer (COC), and features a configuration of micro-pillared array of solid phase multiple extraction beds. The antibody used for exosomes selection was disease specific anti-EpCAM which was covalently attached to microfluidic surface. The design of the chip was computationally optimized using fluidic resistance and diffusion-based capture modelling. The embossing master will be fabricated with UV-LIGA techniques that allows for replication of very high aspect ratio and narrow features that were design to maximize the recovery of exosomes. From the exosome isolate, we extract RNA which is reverse transcribed for qPCR and mutation screening which gives information indicative of a patient¿¿¿s disease state.