Quantification and immunoprofiling of bladder cancer cell-derived extracellular vesicles with microfluidic chemiluminescent ELISA
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...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-09-01
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| Series: | Biosensors and Bioelectronics: X |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2590137021000029 |
| _version_ | 1819134349752139776 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Xiaotian Tan |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-22T10:01:47Z |
| format | Article |
| id | doaj.art-f0e5c940854b43aa825d015c3ef79244 |
| institution | Directory Open Access Journal |
| issn | 2590-1370 |
| language | English |
| last_indexed | 2024-12-22T10:01:47Z |
| publishDate | 2021-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biosensors and Bioelectronics: X |
| spelling | doaj.art-f0e5c940854b43aa825d015c3ef792442022-12-21T18:30:05ZengElsevierBiosensors and Bioelectronics: X2590-13702021-09-018100066Quantification and immunoprofiling of bladder cancer cell-derived extracellular vesicles with microfluidic chemiluminescent ELISAXiaotian Tan0Kathleen C. Day1Xuzhou Li2Luke J. Broses3Wen Xue4Weishu Wu5William Y. Wang6Ting-Wen Lo7Emma Purcell8Sicong Wang9Yun-Lu Sun10Maung Kyaw Khaing Oo11Brendon M. Baker12Sunitha Nagrath13Mark L. Day14Xudong Fan15Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USADepartment of Urology, University of Michigan, Ann Arbor, MI, 48109, USA; Rogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI, 48109, USADepartment of Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA; Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI, 48109, USADepartment of Urology, University of Michigan, Ann Arbor, MI, 48109, USA; Rogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI, 48109, USADepartment of Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USADepartment of Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USADepartment of Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USADepartment of Chemical Engineering, University of Michigan, Ann Arbor, MI, 48109, USADepartment of Chemical Engineering, University of Michigan, Ann Arbor, MI, 48109, USADepartment of Mechanical Engineering, University of Michigan, Ann Arbor, MI, 48109, USADepartment of Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USAOptofluidic Bioassay, LLC, Ann Arbor, MI, 48103, USADepartment of Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USARogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI, 48109, USA; Department of Chemical Engineering, University of Michigan, Ann Arbor, MI, 48109, USADepartment 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.Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA; Corresponding author.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.http://www.sciencedirect.com/science/article/pii/S2590137021000029 |
| spellingShingle | 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 Quantification and immunoprofiling of bladder cancer cell-derived extracellular vesicles with microfluidic chemiluminescent ELISA Biosensors and Bioelectronics: X |
| title | Quantification and immunoprofiling of bladder cancer cell-derived extracellular vesicles with microfluidic chemiluminescent ELISA |
| title_full | Quantification and immunoprofiling of bladder cancer cell-derived extracellular vesicles with microfluidic chemiluminescent ELISA |
| title_fullStr | Quantification and immunoprofiling of bladder cancer cell-derived extracellular vesicles with microfluidic chemiluminescent ELISA |
| title_full_unstemmed | Quantification and immunoprofiling of bladder cancer cell-derived extracellular vesicles with microfluidic chemiluminescent ELISA |
| title_short | Quantification and immunoprofiling of bladder cancer cell-derived extracellular vesicles with microfluidic chemiluminescent ELISA |
| title_sort | quantification and immunoprofiling of bladder cancer cell derived extracellular vesicles with microfluidic chemiluminescent elisa |
| url | http://www.sciencedirect.com/science/article/pii/S2590137021000029 |
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