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Shotgun proteomic analysis using human serum from type 2 diabetes mellitus patients

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International Journal of Diabetes in Developing Countries Aims and scope Submit manuscript

Abstract

Background

Type 2 diabetes mellitus (T2DM), also known as adult-onset diabetes or noninsulin-dependent diabetes mellitus, is characterized by hyperglycemia and insulin resistance. Protein biomarker screening plays an essential role in different diseases. Proteomic methods such as MALDI-TOF based peptide mass fingerprinting, LC-MS/MS based peptide sequencing, and multidimensional liquid phase chromatography (MDLC) coupled with tandem mass spectrometry (MS) shotgun proteomics are used to identify biomarkers.

Methods

In this study, we used a MDLC coupled with tandem MS shotgun proteomic method to demonstrate protein quantitation results by comparing human serum samples from T2DM patients with those of healthy subjects. We utilized quantitative techniques, dimethyl labeling, MDLC by hydrophilic interaction liquid chromatography separated column, and reverse-phase high-performance liquid chromatography coupled with tandem MS to identify proteins with high potential to be T2DM biomarker candidates.

Results

Identified candidates included vitamin D–binding protein, apolipoprotein B-100, apolipoprotein A2, apolipoprotein A1, transthyretin, Ig heavy-chain V–III region BRO, antithrombin-3, fibrinogen gamma chains, fibrinogen alpha chains, and alpha-1-antitrypsin. In addition, we also generated relative protein networks using STRING bioinformatic software.

Conclusion

These potential biomarker candidates might be verified by further experiments such as an ELISA assay or multiple reaction monitoring MS screening.

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Acknowledgements

We thank the funding support from a grant of the Ministry of Science and Technology (NSC 102-2113-M-037-013-MY2), Taipei, Taiwan, and a grant from the cooperation of the National Sun Yat-Sen University and Kaohsiung Medical University (NSYSUKMU 103-P010). We are thankful for the assistance in tandem MS of the Center for Resources, Research and Development (CRRD) of Kaohsiung Medical University.

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Authors and Affiliations

  1. Department of Biomedical Science and Environmental Biology, College of Life Science, Kaohsiung Medical University, Kaohsiung, Taiwan

    Ruei-Nian Li

  2. Cancer Center, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan

    Ruei-Nian Li

  3. Protein Chemistry Core Laboratory, Core Instrument Center, National Health Research Institutes, Miaoli, Taiwan

    Po-Tsun Shen

  4. Graduate Institute of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

    Hugo You-Hsien Lin

  5. Division of Nephrology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan

    Hugo You-Hsien Lin

  6. Department of Internal Medicine, Kaohsiung Municipal Ta-Tung Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan

    Hugo You-Hsien Lin

  7. Department of Biotechnology, College of Life Science, Kaohsiung Medical University, 100, Shih-Chuan 1st Road, Kaohsiung, 80708, Taiwan

    Shih-Shin Liang

  8. Institute of Biomedical Science, College of Science, National Sun Yat-sen University, Kaohsiung, Taiwan

    Shih-Shin Liang

  9. Department of Medical Research, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan

    Shih-Shin Liang

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  1. Ruei-Nian Li
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  2. Po-Tsun Shen
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Correspondence to Shih-Shin Liang.

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The authors declare no competing interests.

Ethical Approval

All serum samples and procedures were approved by the clinical research ethics committee at Kaohsiung Medical University Hospital.

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Li, RN., Shen, PT., Lin, H.YH. et al. Shotgun proteomic analysis using human serum from type 2 diabetes mellitus patients. Int J Diabetes Dev Ctries 43, 145–154 (2023). https://doi.org/10.1007/s13410-021-01038-z

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  • DOI: https://doi.org/10.1007/s13410-021-01038-z

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