# Two-Dimensional Multi-Stage Isotachophoretic Technology for Multiplex Analysis of Cancer Exosomes and Proteins Marker Panel

> **NIH NIH R21** · WASHINGTON STATE UNIVERSITY · 2021 · $207,968

## Abstract

Abstract
Exosomes are small-sized (30–120 nm) extracellular vesicles. They are secreted by most cell types and play
important roles in extracellular communication in normal and pathological processes. The exosomes derived
from cancers shuttle signaling molecules (e.g. proteins and miRNAs) from parental cancer cells and tissue to
distal recipient cells to reprogram the recipient cells and promote tumor growth and metastasis. Therefore,
circulating tumor-derived exosomes carrying signature protein markers of the tumor hold great potential as
invaluable liquid biopsy tools for the noninvasive diagnosis of early-stage cancers. Despite their potential clinical
significance, translating disease-derived exosomes into point-of-care (POC) applications for the early diagnosis
of cancers is hampered by a critical technical barrier: lack of a cost-effective POC approach capable of the
simultaneous analysis of specific exosomes and their content markers in clinical samples. Although a number of
methods for exosome isolation and characterization have been developed, either they are singly functionalized,
nonspecific, laborious, or time-consuming, or they lack the robustness to be adopted as a cost-effective POC
technique. Therefore, there is an urgent need for an effective, precise, easy to use, low-cost approach to
multiplex POC sample analysis to detect trace levels of specific populations of exosomes released by specific
cancer cells at an early stage and comprehensively profile the cancer markers carried by the exosomes. This
application aims to fill the gap by taking a multidisciplinary approach to developing a novel, disposable two-
dimensional paper-based multistage isotachophoresis (ITP) technology platform capable of the simultaneous
analysis of specific target exosomes and exosomal proteins in a cost-effective way. Our objective is to develop
an integrated paper-based isotachophoretic platform on which: 1) anionic cascade ITP is used to deplete high-
abundance plasma proteins and enrich target exosomes before their capture and analysis; 2) a second ITP
process simultaneously analyzes multiplex exosomal proteins released by lysing the exosomes captured in the
first ITP process; and 3) a miniaturized smartphone-based detection module quantifies the target exosomes and
protein markers captured by novel graded-binding test lines. We expect that integrating effective
isolation/identification of specific exosomes with multiplex analysis of their contents in a cost-effective modular
platform will provide a robust POC approach to advancing basic and clinical translational research on disease-
derived exosomes. Exosomes derived from breast cancer will be used as model targets to validate our
technology. The success of this project will not only provide a clinically compatible POC tool for tracking specific
exosomes and markers for early screening for cancers, but also prompt research on the profile analysis of
exosomal markers for the precision diagnosis of o...

## Key facts

- **NIH application ID:** 10119139
- **Project number:** 1R21CA248734-01A1
- **Recipient organization:** WASHINGTON STATE UNIVERSITY
- **Principal Investigator:** WEN-JI DONG
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $207,968
- **Award type:** 1
- **Project period:** 2021-01-01 → 2022-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10119139

## Citation

> US National Institutes of Health, RePORTER application 10119139, Two-Dimensional Multi-Stage Isotachophoretic Technology for Multiplex Analysis of Cancer Exosomes and Proteins Marker Panel (1R21CA248734-01A1). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10119139. Licensed CC0.

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