Open Access iconOpen Access

ARTICLE

A UHPLC/MS/MS Assay Based on an Isotope-Labeled Peptide for Sensitive miR-21 Detection in HCC Serum

Xinyue Wang1,#, Jing Xu1,#, Qihong Gu1, Dingxuan Tang1, Huoyan Ji2, Shaoqing Ju2, Feng Wang2,*, Lin Chen3, Ruoyu Yuan2,*

1 Department of Laboratory Medicine, School of Public Health, Nantong University, Nantong, 226001, China
2 Department of Clinical Laboratory, Affiliated Hospital of Nantong University, Nantong, 226001, China
3 Department of Hepatology Laboratory, Nantong Third Hospital Affiliated to Nantong University, Nantong, 226006, China

* Corresponding Authors: Feng Wang. Email: email; Ruoyu Yuan. Email: email
# These authors contributed equally to this work.

Oncologie 2022, 24(3), 513-526. https://doi.org/10.32604/oncologie.2022.024373

Abstract

Background: MicroRNAs (miRNAs) have been identified as promising novel biomarkers for cancer diagnosis and prognosis, especially for hepatocellular carcinoma (HCC). Nowadays, the expression level of miR-21 in serum samples is a diagnostic indicator for HCC diagnosis. Thus, the quantitative determination of miRNA concentration is of significance in clinical practice. It is particularly important to establish an analytical detection method for miR-21 in patient serum. Methods: The signal readout for miR-21 was based on the mass response of a reporter peptide using an isotope dilution mass spectrometry (MS) method in this work. To be more specific, miR-21 was biotinylated before being coupled with streptavidin (SA) agarose and then hybridized with a newly synthesized DNA-peptide probe. The release and purification of the sample was based on the method including trypsin digestion, solid-phase extraction, and drying, and the detection of the reporter peptide was carried out by UHPLC/MS/MS. The miR-21 in the corresponding samples was quantified by the ratio of the chromatographic peak area of the redissolved polypeptide to that of the isotope-labeled polypeptide. Additionally, within the calibration range, the performance of the method (including precision, accuracy, linearity, and recovery) was evaluated. Results: The concentration of miR-21 was determined using the ratio of relative peak area of stable isotope-labeled internal standard and reporter peptide, yielding a linear range of 0.1∼30.0 nM (y = 0.0818x + 0.7554, R2 = 0.9586, P < 0.01). The limit of detection (LOD) for miR-21 was 10 pM. For y5 , the recoveries (n = 3) were 91.36 ± 2.19%, 93.64 ± 3.55%, and 96.04 ± 2.02% for the levels of three miR-21 samples including RL, RM, and RH, respectively. Conclusions: Overall, this research provides a novel analytical approach for quantitative detection of miRNAs in clinical serum samples.

Keywords


Cite This Article

APA Style
Wang, X., Xu, J., Gu, Q., Tang, D., Ji, H. et al. (2022). A UHPLC/MS/MS assay based on an isotope-labeled peptide for sensitive mir-21 detection in HCC serum. Oncologie, 24(3), 513-526. https://doi.org/10.32604/oncologie.2022.024373
Vancouver Style
Wang X, Xu J, Gu Q, Tang D, Ji H, Ju S, et al. A UHPLC/MS/MS assay based on an isotope-labeled peptide for sensitive mir-21 detection in HCC serum. Oncologie . 2022;24(3):513-526 https://doi.org/10.32604/oncologie.2022.024373
IEEE Style
X. Wang et al., "A UHPLC/MS/MS Assay Based on an Isotope-Labeled Peptide for Sensitive miR-21 Detection in HCC Serum," Oncologie , vol. 24, no. 3, pp. 513-526. 2022. https://doi.org/10.32604/oncologie.2022.024373



cc This work is licensed under a Creative Commons Attribution 4.0 International License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
  • 1078

    View

  • 620

    Download

  • 0

    Like

Share Link