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An LC–MS/MS method for serum cystatin C quantification and its comparison with two commercial immunoassays

  • Li Zhang , Xiaerbanu Nizhamuding , Hao Zheng , Jie Zeng , Xinyi Yuan , Zijia Ma , Weiyan Zhou ORCID logo , Chao Zhang , Tianjiao Zhang EMAIL logo and Chuanbao Zhang ORCID logo EMAIL logo
Published/Copyright: January 23, 2024
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 62 Issue 6

Abstract

Objectives

The standardization of cystatin C (CysC) measurement has received increasing attention in recent years due to its importance in estimating glomerular filtration rate (GFR). Mass spectrometry-based assays have the potential to provide an accuracy base for CysC measurement. However, a precise, accurate and sustainable LC–MS/MS method for CysC is still lacking.

Methods

The developed LC–MS/MS method quantified CysC by detecting signature peptide (T3) obtained from tryptic digestion. Stable isotope labeled T3 peptide (SIL-T3) was spiked to control matrix effects and errors caused by liquid handling. The protein denaturation, reduction and alkylation procedures were combined into a single step with incubation time of 1 h, and the digestion lasted for 3.5 h. In the method validation, digestion time-course, imprecision, accuracy, matrix effect, interference, limit of quantification (LOQ), carryover, linearity, and the comparability to two routine immunoassays were evaluated.

Results

No significant matrix effect or interference was observed with the CysC measurement. The LOQ was 0.21 mg/L; the within-run and total imprecision were 1.33–2.05 % and 2.18–3.90 % for three serum pools (1.18–5.34 mg/L). The LC–MS/MS method was calibrated by ERM-DA471/IFCC and showed good correlation with two immunoassays traceable to ERM-DA471/IFCC. However, significant bias was observed for immunoassays against the LC–MS/MS method.

Conclusions

The developed LC–MS/MS method is robust and simpler and holds the promise to provide an accuracy base for routine immunoassays, which will promote the standardization of CysC measurement.

Keywords: cystatin C; LC–MS/MS; isotope dilution; method validation; reference method; standardization
Corresponding authors: Tianjiao Zhang and Chuanbao Zhang, National Center for Clinical Laboratories, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing Hospital, Beijing, P.R. China; and National Center of Gerontology, Beijing Engineering Research Center of Laboratory Medicine, Beijing, P.R. China, Phone: +86 010 58115059, Fax: +86 010 65132968, E-mail: (T. Zhang), (C. Zhang)

Funding source: National Key Research and Development Program of China

Award Identifier / Grant number: 2022YFC3602301

Funding source: Beijing Natural Science Foundation

Award Identifier / Grant number: L234058

Acknowledgments

We appreciate the technical support of engineers from Roche and DiaSys.

  1. Research ethics: The Ethics Committee of Beijing Hospital approved this study (2022BJYYEC-369-01).

  2. Informed consent: The Ethics Committee of Beijing Hospital exempted the need for obtaining informed consent (2022BJYYEC-369-01).

  3. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission. Li Zhang conceived the study, designed and implemented the experiment, performed data analysis, and drafted the manuscript. Xiaerbanu Nizhamuding, Hao Zheng and Jie Zeng reviewed the literature, participated in experiment design and provided critical revisions. Xinyi Yuan and Zijia Ma conducted the statistical analysis and contributed to the experimental design. Weiyan Zhou and Chao Zhang contributed to experimental design and interpretation of the results. Tianjiao Zhang and Chuanbao Zhang administrated the project, acquired funding and provided final approval of the manuscript for submission.

  4. Competing interests: The authors declare that they have no competing interests.

  5. Research funding: National Key Research and Development Program of China: No. 2022YFC3602301. Beijing Natural Science Foundation: No. L234058.

  6. Data availability: The raw data can be obtained on request from the corresponding author.

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Supplementary Material

This article contains supplementary material (https://doi.org/10.1515/cclm-2023-0821).

Received: 2023-08-01
Accepted: 2024-01-08
Published Online: 2024-01-23
Published in Print: 2024-05-27

© 2024 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2023-0821
Keywords for this article
cystatin C; LC–MS/MS; isotope dilution; method validation; reference method; standardization

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  2. Editorial
  3. SARS-CoV-2 is here to stay: do not lower our guard
  4. Reviews
  5. SARS-CoV-2 subgenomic RNA: formation process and rapid molecular diagnostic methods
  6. Prognostic value of anti-SARS-CoV-2 antibodies: a systematic review
  7. Presence of SARS-CoV-2 RNA in COVID-19 survivors with post-COVID symptoms: a systematic review of the literature
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  13. Report from the HarmoSter study: different LC-MS/MS androstenedione, DHEAS and testosterone methods compare well; however, unifying calibration is a double-edged sword
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  15. CX3CL1/Fractalkine as a biomarker for early pregnancy prediction of preterm premature rupture of membranes
  16. Elevated S100B urine levels predict seizures in infants complicated by perinatal asphyxia and undergoing therapeutic hypothermia
  17. The correlation of urea and creatinine concentrations in sweat and saliva with plasma during hemodialysis: an observational cohort study
  18. Tubular phosphate transport: a comparison between different methods of urine sample collection in FGF23-dependent hypophosphatemic syndromes
  19. Reference Values and Biological Variations
  20. Monocyte distribution width (MDW): study of reference values in blood donors
  21. Data mining of reference intervals for serum creatinine: an improvement in glomerular filtration rate estimating equations based on Q-values
  22. Hematology and Coagulation
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