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Validation of a new method by nano-liquid chromatography on chip tandem mass spectrometry for combined quantitation of C3f and the V65 vitronectin fragment as biomarkers of diagnosis and severity of osteoarthritis

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Validation of a new method by nano-liquid chromatography on chip tandem mass spectrometry for combined quantitation of C3f and the V65 vitronectin fragment as biomarkers of diagnosis and severity of osteoarthritis. / Cobraiville, Gaël; Fillet, Marianne; Sharif, Mohammed; Ourradi, Khadija; Nys, Gwenaël; Malaise, Michel G.; de Seny, Dominique.

In: Talanta, Vol. 169, 01.07.2017, p. 170-180.

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@article{6e6c9e697cc946d483a8867fb2c572d4,
title = "Validation of a new method by nano-liquid chromatography on chip tandem mass spectrometry for combined quantitation of C3f and the V65 vitronectin fragment as biomarkers of diagnosis and severity of osteoarthritis",
abstract = "Microfluidic liquid chromatography coupled to a nanoelectrospray source ion trap mass spectrometry was used for the absolute and simultaneous quantitation of C3f and the V65 vitronectin fragment in serum. The method was first carefully optimized and then validated in serum biological matrix. Stable isotopes for the two biomarkers of interest were used as stable isotope labeled peptide standards. A weighted 1/x2 quadratic regression for C3f and a weighted 1/x quadratic regression for the V65 vitronectin peptide were selected for calibration curves. Trueness (with a relative bias <10{\%}), precision (repeatability and intermediate precision <15{\%}) and accuracy (risk <15{\%}) of the method were successfully demonstrated. The linearity of results was validated in the concentration range of 2.5–200 ng/mL for C3f and 2.5–100 ng/mL for the V65 vitronectin fragment. Serum samples (n=147) classified in 7 groups [(healthy volunteers, OA with 5 grades of severity and rheumatoid arthritis (RA) patients] were analyzed with our new quantitative method. Our data confirm that C3f and the V65 vitronectin fragment are biomarkers of OA severity, but also that C3f fragment is further related to OA severity whereas the V65 vitronectin fragment is more related to early OA detection.",
keywords = "liquid chromatography-chip, mass spectrometry, peptide quantitation, complement C3, vitronectin, validation, serum",
author = "Ga{\"e}l Cobraiville and Marianne Fillet and Mohammed Sharif and Khadija Ourradi and Gwena{\"e}l Nys and Malaise, {Michel G.} and {de Seny}, Dominique",
year = "2017",
month = "7",
day = "1",
doi = "10.1016/j.talanta.2017.03.078",
language = "English",
volume = "169",
pages = "170--180",
journal = "Talanta",
issn = "0039-9140",
publisher = "Amsterdam:Elsevier",

}

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TY - JOUR

T1 - Validation of a new method by nano-liquid chromatography on chip tandem mass spectrometry for combined quantitation of C3f and the V65 vitronectin fragment as biomarkers of diagnosis and severity of osteoarthritis

AU - Cobraiville, Gaël

AU - Fillet, Marianne

AU - Sharif, Mohammed

AU - Ourradi, Khadija

AU - Nys, Gwenaël

AU - Malaise, Michel G.

AU - de Seny, Dominique

PY - 2017/7/1

Y1 - 2017/7/1

N2 - Microfluidic liquid chromatography coupled to a nanoelectrospray source ion trap mass spectrometry was used for the absolute and simultaneous quantitation of C3f and the V65 vitronectin fragment in serum. The method was first carefully optimized and then validated in serum biological matrix. Stable isotopes for the two biomarkers of interest were used as stable isotope labeled peptide standards. A weighted 1/x2 quadratic regression for C3f and a weighted 1/x quadratic regression for the V65 vitronectin peptide were selected for calibration curves. Trueness (with a relative bias <10%), precision (repeatability and intermediate precision <15%) and accuracy (risk <15%) of the method were successfully demonstrated. The linearity of results was validated in the concentration range of 2.5–200 ng/mL for C3f and 2.5–100 ng/mL for the V65 vitronectin fragment. Serum samples (n=147) classified in 7 groups [(healthy volunteers, OA with 5 grades of severity and rheumatoid arthritis (RA) patients] were analyzed with our new quantitative method. Our data confirm that C3f and the V65 vitronectin fragment are biomarkers of OA severity, but also that C3f fragment is further related to OA severity whereas the V65 vitronectin fragment is more related to early OA detection.

AB - Microfluidic liquid chromatography coupled to a nanoelectrospray source ion trap mass spectrometry was used for the absolute and simultaneous quantitation of C3f and the V65 vitronectin fragment in serum. The method was first carefully optimized and then validated in serum biological matrix. Stable isotopes for the two biomarkers of interest were used as stable isotope labeled peptide standards. A weighted 1/x2 quadratic regression for C3f and a weighted 1/x quadratic regression for the V65 vitronectin peptide were selected for calibration curves. Trueness (with a relative bias <10%), precision (repeatability and intermediate precision <15%) and accuracy (risk <15%) of the method were successfully demonstrated. The linearity of results was validated in the concentration range of 2.5–200 ng/mL for C3f and 2.5–100 ng/mL for the V65 vitronectin fragment. Serum samples (n=147) classified in 7 groups [(healthy volunteers, OA with 5 grades of severity and rheumatoid arthritis (RA) patients] were analyzed with our new quantitative method. Our data confirm that C3f and the V65 vitronectin fragment are biomarkers of OA severity, but also that C3f fragment is further related to OA severity whereas the V65 vitronectin fragment is more related to early OA detection.

KW - liquid chromatography-chip

KW - mass spectrometry

KW - peptide quantitation

KW - complement C3

KW - vitronectin

KW - validation

KW - serum

U2 - 10.1016/j.talanta.2017.03.078

DO - 10.1016/j.talanta.2017.03.078

M3 - Article

VL - 169

SP - 170

EP - 180

JO - Talanta

JF - Talanta

SN - 0039-9140

ER -