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Cross-Validation of a Multiplex LC-MS/MS Method for Assaying mAbs Plasma Levels in Patients with Cancer: A GPCO-UNICANCER Study

Background: Different liquid chromatography tandem mass spectrometry (LC–MS/MS) methods have been published for quantification of monoclonal antibodies (mAbs) in plasma but thus far none allowed the simultaneous quantification of several mAbs, including immune checkpoint inhibitors. We developed and...

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Autores principales: Marin, Clémence, Khoudour, Nihel, Millet, Aurélien, Lebert, Dorothée, Bros, Pauline, Thomas, Fabienne, Ternant, David, Lacarelle, Bruno, Guitton, Jérôme, Ciccolini, Joseph, Blanchet, Benoit
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8401780/
https://www.ncbi.nlm.nih.gov/pubmed/34451893
http://dx.doi.org/10.3390/ph14080796
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author Marin, Clémence
Khoudour, Nihel
Millet, Aurélien
Lebert, Dorothée
Bros, Pauline
Thomas, Fabienne
Ternant, David
Lacarelle, Bruno
Guitton, Jérôme
Ciccolini, Joseph
Blanchet, Benoit
author_facet Marin, Clémence
Khoudour, Nihel
Millet, Aurélien
Lebert, Dorothée
Bros, Pauline
Thomas, Fabienne
Ternant, David
Lacarelle, Bruno
Guitton, Jérôme
Ciccolini, Joseph
Blanchet, Benoit
author_sort Marin, Clémence
collection PubMed
description Background: Different liquid chromatography tandem mass spectrometry (LC–MS/MS) methods have been published for quantification of monoclonal antibodies (mAbs) in plasma but thus far none allowed the simultaneous quantification of several mAbs, including immune checkpoint inhibitors. We developed and validated an original multiplex LC–MS/MS method using a ready-to-use kit to simultaneously assay 7 mAbs (i.e., bevacizumab, cetuximab, ipilimumab, nivolumab, pembrolizumab, rituximab and trastuzumab) in plasma. This method was next cross-validated with respective reference methods (ELISA or LC–MS/MS). Methods: The mAbXmise kit was used for mAb extraction and full-length stable-isotope-labeled antibodies as internal standards. The LC–MS/MS method was fully validated following current EMA guidelines. Each cross validation between reference methods and ours included 16–28 plasma samples from cancer patients. Results: The method was linear from 2 to 100 µg/mL for all mAbs. Inter- and intra-assay precision was <14.6% and accuracy was 90.1–111.1%. The mean absolute bias of measured concentrations between multiplex and reference methods was 10.6% (range 3.0–19.9%). Conclusions: We developed and cross-validated a simple, accurate and precise method that allows the assay of up to 7 mAbs. Furthermore, the present method is the first to offer a simultaneous quantification of three immune checkpoint inhibitors likely to be associated in patients.
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spelling pubmed-84017802021-08-29 Cross-Validation of a Multiplex LC-MS/MS Method for Assaying mAbs Plasma Levels in Patients with Cancer: A GPCO-UNICANCER Study Marin, Clémence Khoudour, Nihel Millet, Aurélien Lebert, Dorothée Bros, Pauline Thomas, Fabienne Ternant, David Lacarelle, Bruno Guitton, Jérôme Ciccolini, Joseph Blanchet, Benoit Pharmaceuticals (Basel) Article Background: Different liquid chromatography tandem mass spectrometry (LC–MS/MS) methods have been published for quantification of monoclonal antibodies (mAbs) in plasma but thus far none allowed the simultaneous quantification of several mAbs, including immune checkpoint inhibitors. We developed and validated an original multiplex LC–MS/MS method using a ready-to-use kit to simultaneously assay 7 mAbs (i.e., bevacizumab, cetuximab, ipilimumab, nivolumab, pembrolizumab, rituximab and trastuzumab) in plasma. This method was next cross-validated with respective reference methods (ELISA or LC–MS/MS). Methods: The mAbXmise kit was used for mAb extraction and full-length stable-isotope-labeled antibodies as internal standards. The LC–MS/MS method was fully validated following current EMA guidelines. Each cross validation between reference methods and ours included 16–28 plasma samples from cancer patients. Results: The method was linear from 2 to 100 µg/mL for all mAbs. Inter- and intra-assay precision was <14.6% and accuracy was 90.1–111.1%. The mean absolute bias of measured concentrations between multiplex and reference methods was 10.6% (range 3.0–19.9%). Conclusions: We developed and cross-validated a simple, accurate and precise method that allows the assay of up to 7 mAbs. Furthermore, the present method is the first to offer a simultaneous quantification of three immune checkpoint inhibitors likely to be associated in patients. MDPI 2021-08-12 /pmc/articles/PMC8401780/ /pubmed/34451893 http://dx.doi.org/10.3390/ph14080796 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Marin, Clémence
Khoudour, Nihel
Millet, Aurélien
Lebert, Dorothée
Bros, Pauline
Thomas, Fabienne
Ternant, David
Lacarelle, Bruno
Guitton, Jérôme
Ciccolini, Joseph
Blanchet, Benoit
Cross-Validation of a Multiplex LC-MS/MS Method for Assaying mAbs Plasma Levels in Patients with Cancer: A GPCO-UNICANCER Study
title Cross-Validation of a Multiplex LC-MS/MS Method for Assaying mAbs Plasma Levels in Patients with Cancer: A GPCO-UNICANCER Study
title_full Cross-Validation of a Multiplex LC-MS/MS Method for Assaying mAbs Plasma Levels in Patients with Cancer: A GPCO-UNICANCER Study
title_fullStr Cross-Validation of a Multiplex LC-MS/MS Method for Assaying mAbs Plasma Levels in Patients with Cancer: A GPCO-UNICANCER Study
title_full_unstemmed Cross-Validation of a Multiplex LC-MS/MS Method for Assaying mAbs Plasma Levels in Patients with Cancer: A GPCO-UNICANCER Study
title_short Cross-Validation of a Multiplex LC-MS/MS Method for Assaying mAbs Plasma Levels in Patients with Cancer: A GPCO-UNICANCER Study
title_sort cross-validation of a multiplex lc-ms/ms method for assaying mabs plasma levels in patients with cancer: a gpco-unicancer study
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8401780/
https://www.ncbi.nlm.nih.gov/pubmed/34451893
http://dx.doi.org/10.3390/ph14080796
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