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Development of commutable candidate certified reference materials from protein solutions: concept and application to human insulin

  • Shiwen Luo , Yao Tan , Ziliang Wang , Bin Yang , Yahui Liu , Jing Wang and Liqing Wu EMAIL logo
Published/Copyright: April 24, 2025
Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 63 Issue 9

Abstract

Objectives

To explore the reasons behind the lack of commutability in protein solution reference materials (RMs) and seeks to eliminate these factors in order to develop commutable reference materials, using human insulin (hINS) as example, to meet the growing demand for standardization in in vitro diagnostics.

Methods

A concept for development of commutable protein solution RMs by matrix matching, structural-activity analysis, higher order structure adjustment, active concentration measurement, and commutability verification was investigated. This concept was applied in the development of hINS solution candidate certified RMs (cCRMs). Bovine serum solution (7 %) was used for matrix matching and hINS-Zn2+ aggregates were found to be the key for commutability. hINS-Zn2+ aggregates were constructed in vitro and the presence of the aggregates was confirmed through circular dichroism spectroscopy and mass photometry. The active concentration of the aggregate solution was analyzed using surface plasmon resonance. Then, six levels of hINS solution cCRMs were developed and the commutability was evaluated using both CLSI EP14 and IFCC approaches.

Results

The hINS solution cCRMs exhibited excellent homogeneity and remained stable for at least 6 months when stored at −70 °C. The relative uncertainties of these cCRMs ranged from 4.0 to 5.0 %. Both CLSI EP14 and IFCC commutability evaluations indicated good commutability between routine chemiluminescent immunoassay systems.

Conclusions

A new concept for developing commutable RMs with pure protein material, which avoids the challenges in developing commutable matrix RMs and should contribute to the standardization of clinical test results for proteins, was successfully applied to develop commutable hINS solution cCRMs.

Keywords: insulin; commutability; reference material; structure adjustment; protein
Corresponding author: Liqing Wu, National Institute of Metrology, Beijing, 100029, China, E-mail:

Funding source: the Key R&D Plan of the Ministry of Science and Technology

Award Identifier / Grant number: 2021YFF0600802

Funding source: Biological Breeding-National Science and Technology Major Project

Award Identifier / Grant number: 2022ZD0402007

Acknowledgment

Victoria Muir, PhD, from Liwen Bianji, (Edanz) (www.liwenbianji.cn/), edited the English text of a draft of this manuscript.

  1. Research ethics: Not applicable.

  2. Informed consent: Not applicable.

  3. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  4. Use of Large Language Models, AI and Machine Learning Tools: None declared.

  5. Conflict of interest: The authors state no conflict of interest.

  6. Research funding: This work was supported by the Key R&D Plan of the Ministry of Science and Technology with grant no. 2021YFF0600802, and Biological Breeding-National Science and Technology Major Project with grant no. 2022ZD0402007.

  7. 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-2025-0329).

Received: 2025-03-17
Accepted: 2025-04-14
Published Online: 2025-04-24
Published in Print: 2025-08-26

© 2025 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2025-0329
Keywords for this article
insulin; commutability; reference material; structure adjustment; protein

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Macroprolactinaemia – some progress but still an ongoing problem
  4. Review
  5. Understanding the circulating forms of cardiac troponin: insights for clinical practice
  6. Opinion Papers
  7. New insights in preanalytical quality
  8. IFCC recommendations for internal quality control practice: a missed opportunity
  9. Genetics and Molecular Diagnostics
  10. Evaluation of error detection and treatment recommendations in nucleic acid test reports using ChatGPT models
  11. General Clinical Chemistry and Laboratory Medicine
  12. Pre-analytical phase errors constitute the vast majority of errors in clinical laboratory testing
  13. Improving the efficiency of quality control in clinical laboratory with an integrated PBRTQC system based on patient risk
  14. IgA-type macroprolactin among 130 patients with macroprolactinemia
  15. Prevalence and re-evaluation of macroprolactinemia in hyperprolactinemic patients: a retrospective study in the Turkish population
  16. Defining dried blood spot diameter: implications for measurement and specimen rejection rates
  17. Screening primary aldosteronism by plasma aldosterone-to-angiotensin II ratio
  18. Assessment of serum free light chain measurements in a large Chinese chronic kidney disease cohort: a multicenter real-world study
  19. Beyond the Hydrashift assay: the utility of isoelectric focusing for therapeutic antibody and paraprotein detection
  20. Direct screening and quantification of monoclonal immunoglobulins in serum using MALDI-TOF mass spectrometry without antibody enrichment
  21. Effect of long-term frozen storage on stability of kappa free light chain index
  22. Impact of renal function impairment on kappa free light chain index
  23. Standardization challenges in antipsychotic drug monitoring: insights from a national survey in Chinese TDM practices
  24. Potential coeliac disease in children: a single-center experience
  25. Vitamin D metabolome in preterm infants: insights into postnatal metabolism
  26. Candidate Reference Measurement Procedures and Materials
  27. Development of commutable candidate certified reference materials from protein solutions: concept and application to human insulin
  28. Reference Values and Biological Variations
  29. Biological variation of serum cholinesterase activity in healthy subjects
  30. Hematology and Coagulation
  31. Diagnostic performance of morphological analysis and red blood cell parameter-based algorithms in the routine laboratory screening of heterozygous haemoglobinopathies
  32. Cancer Diagnostics
  33. Promising protein biomarkers for early gastric cancer: clinical performance of combined detection
  34. Infectious Diseases
  35. The accuracy of presepsin in diagnosing neonatal late-onset sepsis in critically ill neonates: a prospective study
  36. Corrigendum
  37. The Unholy Grail of cancer screening: or is it just about the Benjamins?
  38. Letters to the Editor
  39. Analytical validation of hemolysis detection on GEM Premier 7000
  40. Reconciling reference ranges and clinical decision limits: the case of thyroid stimulating hormone
  41. Contradictory definitions give rise to demands for a right to unambiguous definitions
  42. Biomarkers to measure the need and the effectiveness of therapeutic supplementation: a critical issue
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