Quantification and immunoprofiling of bladder cancer cell-derived extracellular vesicles with microfluidic chemiluminescent ELISA

Xiaotian Tan; Kathleen C. Day; Xuzhou Li; Luke J. Broses; Wen Xue; Weishu Wu; William Y. Wang; Ting-Wen Lo; Emma Purcell; Sicong Wang; Yun-Lu Sun; Maung Kyaw Khaing Oo; Brendon M. Baker; Sunitha Nagrath; Mark L. Day; Xudong Fan

Biosensors and Bioelectronics: X (Sep 2021)

Quantification and immunoprofiling of bladder cancer cell-derived extracellular vesicles with microfluidic chemiluminescent ELISA

Xiaotian Tan,
Kathleen C. Day,
Xuzhou Li,
Luke J. Broses,
Wen Xue,
Weishu Wu,
William Y. Wang,
Ting-Wen Lo,
Emma Purcell,
Sicong Wang,
Yun-Lu Sun,
Maung Kyaw Khaing Oo,
Brendon M. Baker,
Sunitha Nagrath,
Mark L. Day,
Xudong Fan

Affiliations

Xiaotian Tan: Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA
Kathleen C. Day: Department of Urology, University of Michigan, Ann Arbor, MI, 48109, USA; Rogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI, 48109, USA
Xuzhou Li: Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA; Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA
Luke J. Broses: Department of Urology, University of Michigan, Ann Arbor, MI, 48109, USA; Rogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI, 48109, USA
Wen Xue: Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA
Weishu Wu: Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA
William Y. Wang: Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA
Ting-Wen Lo: Department of Chemical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA
Emma Purcell: Department of Chemical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA
Sicong Wang: Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA
Yun-Lu Sun: Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA
Maung Kyaw Khaing Oo: Optofluidic Bioassay, LLC, Ann Arbor, MI, 48103, USA
Brendon M. Baker: Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA
Sunitha Nagrath: Rogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI, 48109, USA; Department of Chemical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA
Mark L. Day: Department of Urology, University of Michigan, Ann Arbor, MI, 48109, USA; Rogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI, 48109, USA; Corresponding author. Department of Urology, University of Michigan, Ann Arbor, MI, 48109, USA.
Xudong Fan: Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA; Corresponding author.

Journal volume & issue: Vol. 8
p. 100066

Abstract

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The functional membrane proteins on tumor-cell-derived EVs contain a large amount of biomolecular information, and can serve as a comprehensive marker to delineate the molecular nature of cancer. However, due to low secretion rates, it is difficult to perform accurate quantification and biomolecular analysis with conventional EV analysis technologies such as the Western blots. Here, we introduce a multifunctional EV analysis technology based on an automated microfluidic chemiluminescent ELISA (Enzyme-Linked ImmunoSorbent Assay) platform. With this system, we were able to achieve rapid EV quantification (<1 h) with relatively small sample volume (~8 μL) and high sensitivity (optimal LOD = 8.7×107 EV/mL). In addition to the EV quantification, we evaluated the expression levels for a panel of four cancer-related EV membrane proteins (EGFR, HER2, MHC-I, and EpCAM) using a newly developed immunoprofiling assay that combines immunoprecipitation and sandwich ELISA. Due to high sensitivity, this immunoprofiling assay only requires a very small amount of input protein (<40 ng/marker). Our studies show that the expression level of functional EV membrane proteins is stable under external stimulation, which suggests that the expression profile of the EV membrane proteins may serve as a robust and unique “molecular fingerprint” for the immunophenotyping of cancer cell lines.

Published in Biosensors and Bioelectronics: X

ISSN: 2590-1370 (Online)
Publisher: Elsevier
Country of publisher: Netherlands
LCC subjects: Technology: Chemical technology: Biotechnology
Website: https://www.journals.elsevier.com/biosensors-and-bioelectronics-x

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