Automated Extracellular Vesicles Extractor Based on Nanotechnology for High Throughput Sample Preparation and Phosphoproteomics Analysis

Yuchen Yang; Hao Zhang; Yuhan Cai; Yajie Ding; Guiyuan Zhang; Yufeng Liu; Jie Sun; Weiguo Andy Tao; Yanhong Gu

doi:doi:10.11648/j.sd.20221002.15

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Automated Extracellular Vesicles Extractor Based on Nanotechnology for High Throughput Sample Preparation and Phosphoproteomics Analysis

Yuchen Yang, Hao Zhang, Yuhan Cai, Yajie Ding, Guiyuan Zhang, Yufeng Liu, Jie Sun, Weiguo Andy Tao, Yanhong Gu

Published in Science Discovery (Volume 10, Issue 2)

Received: 18 April 2022 Published: 22 April 2022

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Abstract

Extracellular vesicles (EVs) play important roles in the diagnosis and treatment of diseases because of its lavish bioinformatic molecular contents. However, how to separate plentiful of extracellular vesicles in a short time with high consistency is still a problem. Here, we’d like to introduce an automated extracellular vesicles capturing machine named EVenrich (Extracellular Vesicles Enrichment) that helps realize capturing EVs of biosamples in a high-throughput manner. Incorporating EVenrich with EVtraps-a kind of biological magnetic beads capturing extracellular vesicles specifically, we were able to isolate EVs from up to 48 urine samples in 60 min, with similar quality and quantity and superior stability to manual capture. In clinical urine EVs analysis, over 14000 peptide fragments and 2000 proteins were detected by automated capture, which was similar to manual capture and significantly superior to ultracentrifugation. We further collected urine samples from 21 patients with negative prostate biopsy results and 21 patients with confirmed prostate cancer. Every three samples were mixed into a group to make mixed samples of 7 control groups and 7 disease groups. After exosome extraction and exosome proteomics / phosphorylated proteomics analysis, we found 211 overexpressed proteins and 187 overexpressed phosphorylated peptides corresponding to 40 phosphorylated proteins as potential biomarkers for prostate cancer. In addition, qRT-PCR results showed that the ratio of miR-125b to miR-145 could well distinguish prostate cancer patients from non-prostate cancer patients, suggesting that it could be used as a potential marker of prostate cancer.

Published in	Science Discovery (Volume 10, Issue 2)
DOI	10.11648/j.sd.20221002.15
Page(s)	48-59
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2022. Published by Science Publishing Group

Keywords

Extracellular Vesicles, EVenrich, Phosphoproteomics, Prostate Cancer

References

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APA Style

Yuchen Yang, Hao Zhang, Yuhan Cai, Yajie Ding, Guiyuan Zhang, et al. (2022). Automated Extracellular Vesicles Extractor Based on Nanotechnology for High Throughput Sample Preparation and Phosphoproteomics Analysis. Science Discovery, 10(2), 48-59. https://doi.org/10.11648/j.sd.20221002.15

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ACS Style

Yuchen Yang; Hao Zhang; Yuhan Cai; Yajie Ding; Guiyuan Zhang, et al. Automated Extracellular Vesicles Extractor Based on Nanotechnology for High Throughput Sample Preparation and Phosphoproteomics Analysis. Sci. Discov. 2022, 10(2), 48-59. doi: 10.11648/j.sd.20221002.15

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AMA Style

Yuchen Yang, Hao Zhang, Yuhan Cai, Yajie Ding, Guiyuan Zhang, et al. Automated Extracellular Vesicles Extractor Based on Nanotechnology for High Throughput Sample Preparation and Phosphoproteomics Analysis. Sci Discov. 2022;10(2):48-59. doi: 10.11648/j.sd.20221002.15

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@article{10.11648/j.sd.20221002.15,
  author = {Yuchen Yang and Hao Zhang and Yuhan Cai and Yajie Ding and Guiyuan Zhang and Yufeng Liu and Jie Sun and Weiguo Andy Tao and Yanhong Gu},
  title = {Automated Extracellular Vesicles Extractor Based on Nanotechnology for High Throughput Sample Preparation and Phosphoproteomics Analysis},
  journal = {Science Discovery},
  volume = {10},
  number = {2},
  pages = {48-59},
  doi = {10.11648/j.sd.20221002.15},
  url = {https://doi.org/10.11648/j.sd.20221002.15},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.sd.20221002.15},
  abstract = {Extracellular vesicles (EVs) play important roles in the diagnosis and treatment of diseases because of its lavish bioinformatic molecular contents. However, how to separate plentiful of extracellular vesicles in a short time with high consistency is still a problem. Here, we’d like to introduce an automated extracellular vesicles capturing machine named EVenrich (Extracellular Vesicles Enrichment) that helps realize capturing EVs of biosamples in a high-throughput manner. Incorporating EVenrich with EVtraps-a kind of biological magnetic beads capturing extracellular vesicles specifically, we were able to isolate EVs from up to 48 urine samples in 60 min, with similar quality and quantity and superior stability to manual capture. In clinical urine EVs analysis, over 14000 peptide fragments and 2000 proteins were detected by automated capture, which was similar to manual capture and significantly superior to ultracentrifugation. We further collected urine samples from 21 patients with negative prostate biopsy results and 21 patients with confirmed prostate cancer. Every three samples were mixed into a group to make mixed samples of 7 control groups and 7 disease groups. After exosome extraction and exosome proteomics / phosphorylated proteomics analysis, we found 211 overexpressed proteins and 187 overexpressed phosphorylated peptides corresponding to 40 phosphorylated proteins as potential biomarkers for prostate cancer. In addition, qRT-PCR results showed that the ratio of miR-125b to miR-145 could well distinguish prostate cancer patients from non-prostate cancer patients, suggesting that it could be used as a potential marker of prostate cancer.},
 year = {2022}
}

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TY  - JOUR
T1  - Automated Extracellular Vesicles Extractor Based on Nanotechnology for High Throughput Sample Preparation and Phosphoproteomics Analysis
AU  - Yuchen Yang
AU  - Hao Zhang
AU  - Yuhan Cai
AU  - Yajie Ding
AU  - Guiyuan Zhang
AU  - Yufeng Liu
AU  - Jie Sun
AU  - Weiguo Andy Tao
AU  - Yanhong Gu
Y1  - 2022/04/22
PY  - 2022
N1  - https://doi.org/10.11648/j.sd.20221002.15
DO  - 10.11648/j.sd.20221002.15
T2  - Science Discovery
JF  - Science Discovery
JO  - Science Discovery
SP  - 48
EP  - 59
PB  - Science Publishing Group
SN  - 2331-0650
UR  - https://doi.org/10.11648/j.sd.20221002.15
AB  - Extracellular vesicles (EVs) play important roles in the diagnosis and treatment of diseases because of its lavish bioinformatic molecular contents. However, how to separate plentiful of extracellular vesicles in a short time with high consistency is still a problem. Here, we’d like to introduce an automated extracellular vesicles capturing machine named EVenrich (Extracellular Vesicles Enrichment) that helps realize capturing EVs of biosamples in a high-throughput manner. Incorporating EVenrich with EVtraps-a kind of biological magnetic beads capturing extracellular vesicles specifically, we were able to isolate EVs from up to 48 urine samples in 60 min, with similar quality and quantity and superior stability to manual capture. In clinical urine EVs analysis, over 14000 peptide fragments and 2000 proteins were detected by automated capture, which was similar to manual capture and significantly superior to ultracentrifugation. We further collected urine samples from 21 patients with negative prostate biopsy results and 21 patients with confirmed prostate cancer. Every three samples were mixed into a group to make mixed samples of 7 control groups and 7 disease groups. After exosome extraction and exosome proteomics / phosphorylated proteomics analysis, we found 211 overexpressed proteins and 187 overexpressed phosphorylated peptides corresponding to 40 phosphorylated proteins as potential biomarkers for prostate cancer. In addition, qRT-PCR results showed that the ratio of miR-125b to miR-145 could well distinguish prostate cancer patients from non-prostate cancer patients, suggesting that it could be used as a potential marker of prostate cancer.
VL  - 10
IS  - 2
ER  -

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Author Information

Yuchen Yang

Department of Oncology & Cancer Rehabilitation Center, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China
Hao Zhang

State Key Laboratory of Bioelectronics, National Demonstration Center for Experimental Biomedical Engineering Education, Southeast University, Nanjing, China
Yuhan Cai

State Key Laboratory of Bioelectronics, National Demonstration Center for Experimental Biomedical Engineering Education, Southeast University, Nanjing, China
Yajie Ding

State Key Laboratory of Bioelectronics, National Demonstration Center for Experimental Biomedical Engineering Education, Southeast University, Nanjing, China
Guiyuan Zhang

State Key Laboratory of Bioelectronics, National Demonstration Center for Experimental Biomedical Engineering Education, Southeast University, Nanjing, China
Yufeng Liu

State Key Laboratory of Bioelectronics, National Demonstration Center for Experimental Biomedical Engineering Education, Southeast University, Nanjing, China
Jie Sun

State Key Laboratory of Bioelectronics, National Demonstration Center for Experimental Biomedical Engineering Education, Southeast University, Nanjing, China
Weiguo Andy Tao

Department of Chemistry, Department of Biochemistry & Center for Cancer Research, Purdue University, West Lafayette, United States
Yanhong Gu

Department of Oncology & Cancer Rehabilitation Center, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China

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APA Style

Yuchen Yang, Hao Zhang, Yuhan Cai, Yajie Ding, Guiyuan Zhang, et al. (2022). Automated Extracellular Vesicles Extractor Based on Nanotechnology for High Throughput Sample Preparation and Phosphoproteomics Analysis. Science Discovery, 10(2), 48-59. https://doi.org/10.11648/j.sd.20221002.15

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ACS Style

Yuchen Yang; Hao Zhang; Yuhan Cai; Yajie Ding; Guiyuan Zhang, et al. Automated Extracellular Vesicles Extractor Based on Nanotechnology for High Throughput Sample Preparation and Phosphoproteomics Analysis. Sci. Discov. 2022, 10(2), 48-59. doi: 10.11648/j.sd.20221002.15

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AMA Style

Yuchen Yang, Hao Zhang, Yuhan Cai, Yajie Ding, Guiyuan Zhang, et al. Automated Extracellular Vesicles Extractor Based on Nanotechnology for High Throughput Sample Preparation and Phosphoproteomics Analysis. Sci Discov. 2022;10(2):48-59. doi: 10.11648/j.sd.20221002.15

Copy | Download

@article{10.11648/j.sd.20221002.15,
  author = {Yuchen Yang and Hao Zhang and Yuhan Cai and Yajie Ding and Guiyuan Zhang and Yufeng Liu and Jie Sun and Weiguo Andy Tao and Yanhong Gu},
  title = {Automated Extracellular Vesicles Extractor Based on Nanotechnology for High Throughput Sample Preparation and Phosphoproteomics Analysis},
  journal = {Science Discovery},
  volume = {10},
  number = {2},
  pages = {48-59},
  doi = {10.11648/j.sd.20221002.15},
  url = {https://doi.org/10.11648/j.sd.20221002.15},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.sd.20221002.15},
  abstract = {Extracellular vesicles (EVs) play important roles in the diagnosis and treatment of diseases because of its lavish bioinformatic molecular contents. However, how to separate plentiful of extracellular vesicles in a short time with high consistency is still a problem. Here, we’d like to introduce an automated extracellular vesicles capturing machine named EVenrich (Extracellular Vesicles Enrichment) that helps realize capturing EVs of biosamples in a high-throughput manner. Incorporating EVenrich with EVtraps-a kind of biological magnetic beads capturing extracellular vesicles specifically, we were able to isolate EVs from up to 48 urine samples in 60 min, with similar quality and quantity and superior stability to manual capture. In clinical urine EVs analysis, over 14000 peptide fragments and 2000 proteins were detected by automated capture, which was similar to manual capture and significantly superior to ultracentrifugation. We further collected urine samples from 21 patients with negative prostate biopsy results and 21 patients with confirmed prostate cancer. Every three samples were mixed into a group to make mixed samples of 7 control groups and 7 disease groups. After exosome extraction and exosome proteomics / phosphorylated proteomics analysis, we found 211 overexpressed proteins and 187 overexpressed phosphorylated peptides corresponding to 40 phosphorylated proteins as potential biomarkers for prostate cancer. In addition, qRT-PCR results showed that the ratio of miR-125b to miR-145 could well distinguish prostate cancer patients from non-prostate cancer patients, suggesting that it could be used as a potential marker of prostate cancer.},
 year = {2022}
}

Copy | Download

TY  - JOUR
T1  - Automated Extracellular Vesicles Extractor Based on Nanotechnology for High Throughput Sample Preparation and Phosphoproteomics Analysis
AU  - Yuchen Yang
AU  - Hao Zhang
AU  - Yuhan Cai
AU  - Yajie Ding
AU  - Guiyuan Zhang
AU  - Yufeng Liu
AU  - Jie Sun
AU  - Weiguo Andy Tao
AU  - Yanhong Gu
Y1  - 2022/04/22
PY  - 2022
N1  - https://doi.org/10.11648/j.sd.20221002.15
DO  - 10.11648/j.sd.20221002.15
T2  - Science Discovery
JF  - Science Discovery
JO  - Science Discovery
SP  - 48
EP  - 59
PB  - Science Publishing Group
SN  - 2331-0650
UR  - https://doi.org/10.11648/j.sd.20221002.15
AB  - Extracellular vesicles (EVs) play important roles in the diagnosis and treatment of diseases because of its lavish bioinformatic molecular contents. However, how to separate plentiful of extracellular vesicles in a short time with high consistency is still a problem. Here, we’d like to introduce an automated extracellular vesicles capturing machine named EVenrich (Extracellular Vesicles Enrichment) that helps realize capturing EVs of biosamples in a high-throughput manner. Incorporating EVenrich with EVtraps-a kind of biological magnetic beads capturing extracellular vesicles specifically, we were able to isolate EVs from up to 48 urine samples in 60 min, with similar quality and quantity and superior stability to manual capture. In clinical urine EVs analysis, over 14000 peptide fragments and 2000 proteins were detected by automated capture, which was similar to manual capture and significantly superior to ultracentrifugation. We further collected urine samples from 21 patients with negative prostate biopsy results and 21 patients with confirmed prostate cancer. Every three samples were mixed into a group to make mixed samples of 7 control groups and 7 disease groups. After exosome extraction and exosome proteomics / phosphorylated proteomics analysis, we found 211 overexpressed proteins and 187 overexpressed phosphorylated peptides corresponding to 40 phosphorylated proteins as potential biomarkers for prostate cancer. In addition, qRT-PCR results showed that the ratio of miR-125b to miR-145 could well distinguish prostate cancer patients from non-prostate cancer patients, suggesting that it could be used as a potential marker of prostate cancer.
VL  - 10
IS  - 2
ER  -

Copy | Download