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A high-throughput assay for the quantification of intact insulin-like growth factor I in human serum using online SPE-LC-HRMS

Gilles Coppieters (UGent) , Péter Judák (UGent) , Nicolas Van Haecke (UGent) , Pieter Van Renterghem (UGent) , Peter Van Eenoo (UGent) and Koen Deventer (UGent)
(2020) CLINICA CHIMICA ACTA. 510. p.391-399
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Abstract
Quantification of IGF-I is relevant in both doping control as a biomarker of growth hormone (GH) misuse in sports, and in the clinical field for longitudinal follow-up of patients with disorders related to the GH axis. Currently, better standardization of IGF-I measurements using mass spectrometry is in our best interest as it would enable long-term monitoring of an athletes' IGF-I levels by its addition to the Athlete Biological Passport (ABP). Here, a simplified and rapid top-down LC-HAMS method for quantification of IGF-I in human serum is presented. A ten-minute precipitation-based offline sample preparation is combined with online sample clean-up and separation on a conventional LC, resulting in a total runtime of nine minutes in between injections. The method was validated in the relevant range of 50-1000 ng/mL for the following parameters: linearity, precision, bias, Limit Of Quantification (LOQ), carry-over, selectivity, recovery and ion suppression. As proof of concept, the presented LC-HAMS assay was compared with results from a previous inter-laboratory study on intact IGF-I quantification using four human GH administration samples. It was additionally compared with the IDS-iSYS immunoassay using 47 athlete serum samples, showing good overall agreement with a slight positive bias of 24.2 ng/mL for the LC-HAMS assay at a mean sample concentration of 234 ng/mL. Also, a discrepancy between commercially available IGF-I reference material for the calibration of quantitative assays is discussed. This is of importance if LC-MS assays for IGF-I are to be harmonized.
Keywords
MASS-SPECTROMETRY, IGF-I, IMMUNOAFFINITY PURIFICATION, HORMONE ABUSE, PROTEINS, PLASMA, BIOMARKERS, QUANTITATION, AGREEMENT, DISEASE, Insulin-like Growth Factor I (IGF-I), Liquid Chromatography, High-Resolution Mass, Spectrometry (LC-HAMS), Top-down, Quantification, Serum, Doping control

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Citation

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MLA
Coppieters, Gilles, et al. “A High-Throughput Assay for the Quantification of Intact Insulin-like Growth Factor I in Human Serum Using Online SPE-LC-HRMS.” CLINICA CHIMICA ACTA, vol. 510, 2020, pp. 391–99, doi:10.1016/j.cca.2020.07.054.
APA
Coppieters, G., Judák, P., Van Haecke, N., Van Renterghem, P., Van Eenoo, P., & Deventer, K. (2020). A high-throughput assay for the quantification of intact insulin-like growth factor I in human serum using online SPE-LC-HRMS. CLINICA CHIMICA ACTA, 510, 391–399. https://doi.org/10.1016/j.cca.2020.07.054
Chicago author-date
Coppieters, Gilles, Péter Judák, Nicolas Van Haecke, Pieter Van Renterghem, Peter Van Eenoo, and Koen Deventer. 2020. “A High-Throughput Assay for the Quantification of Intact Insulin-like Growth Factor I in Human Serum Using Online SPE-LC-HRMS.” CLINICA CHIMICA ACTA 510: 391–99. https://doi.org/10.1016/j.cca.2020.07.054.
Chicago author-date (all authors)
Coppieters, Gilles, Péter Judák, Nicolas Van Haecke, Pieter Van Renterghem, Peter Van Eenoo, and Koen Deventer. 2020. “A High-Throughput Assay for the Quantification of Intact Insulin-like Growth Factor I in Human Serum Using Online SPE-LC-HRMS.” CLINICA CHIMICA ACTA 510: 391–399. doi:10.1016/j.cca.2020.07.054.
Vancouver
1.
Coppieters G, Judák P, Van Haecke N, Van Renterghem P, Van Eenoo P, Deventer K. A high-throughput assay for the quantification of intact insulin-like growth factor I in human serum using online SPE-LC-HRMS. CLINICA CHIMICA ACTA. 2020;510:391–9.
IEEE
[1]
G. Coppieters, P. Judák, N. Van Haecke, P. Van Renterghem, P. Van Eenoo, and K. Deventer, “A high-throughput assay for the quantification of intact insulin-like growth factor I in human serum using online SPE-LC-HRMS,” CLINICA CHIMICA ACTA, vol. 510, pp. 391–399, 2020.
@article{8686577,
  abstract     = {{Quantification of IGF-I is relevant in both doping control as a biomarker of growth hormone (GH) misuse in sports, and in the clinical field for longitudinal follow-up of patients with disorders related to the GH axis. Currently, better standardization of IGF-I measurements using mass spectrometry is in our best interest as it would enable long-term monitoring of an athletes' IGF-I levels by its addition to the Athlete Biological Passport (ABP). Here, a simplified and rapid top-down LC-HAMS method for quantification of IGF-I in human serum is presented. A ten-minute precipitation-based offline sample preparation is combined with online sample clean-up and separation on a conventional LC, resulting in a total runtime of nine minutes in between injections. The method was validated in the relevant range of 50-1000 ng/mL for the following parameters: linearity, precision, bias, Limit Of Quantification (LOQ), carry-over, selectivity, recovery and ion suppression. As proof of concept, the presented LC-HAMS assay was compared with results from a previous inter-laboratory study on intact IGF-I quantification using four human GH administration samples. It was additionally compared with the IDS-iSYS immunoassay using 47 athlete serum samples, showing good overall agreement with a slight positive bias of 24.2 ng/mL for the LC-HAMS assay at a mean sample concentration of 234 ng/mL. Also, a discrepancy between commercially available IGF-I reference material for the calibration of quantitative assays is discussed. This is of importance if LC-MS assays for IGF-I are to be harmonized.}},
  author       = {{Coppieters, Gilles and Judák, Péter and Van Haecke, Nicolas and Van Renterghem, Pieter and Van Eenoo, Peter and Deventer, Koen}},
  issn         = {{0009-8981}},
  journal      = {{CLINICA CHIMICA ACTA}},
  keywords     = {{MASS-SPECTROMETRY,IGF-I,IMMUNOAFFINITY PURIFICATION,HORMONE ABUSE,PROTEINS,PLASMA,BIOMARKERS,QUANTITATION,AGREEMENT,DISEASE,Insulin-like Growth Factor I (IGF-I),Liquid Chromatography,High-Resolution Mass,Spectrometry (LC-HAMS),Top-down,Quantification,Serum,Doping control}},
  language     = {{eng}},
  pages        = {{391--399}},
  title        = {{A high-throughput assay for the quantification of intact insulin-like growth factor I in human serum using online SPE-LC-HRMS}},
  url          = {{http://dx.doi.org/10.1016/j.cca.2020.07.054}},
  volume       = {{510}},
  year         = {{2020}},
}
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