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A new reliable, transposable and cost-effective assay for absolute quantification of total plasmatic bevacizumab by LC MS/MS in human plasma comparing two internal standard calibration approaches

hal.structure.identifierBiologie des maladies cardiovasculaires = Biology of Cardiovascular Diseases
dc.contributor.authorLEGERON, Rachel
hal.structure.identifierBiologie des maladies cardiovasculaires = Biology of Cardiovascular Diseases
dc.contributor.authorXUEREB, Fabien
hal.structure.identifierChimie et Biologie des Membranes et des Nanoobjets [CBMN]
dc.contributor.authorCHAIGNEPAIN, Stephane
hal.structure.identifierBiologie des maladies cardiovasculaires = Biology of Cardiovascular Diseases
dc.contributor.authorGADEAU, Alain-Pierre
hal.structure.identifierChimie et Biologie des Membranes et des Nanoobjets [CBMN]
dc.contributor.authorCLAVEROL, Stephane
hal.structure.identifierChimie et Biologie des Membranes et des Nanoobjets [CBMN]
dc.contributor.authorDUPUY, Jean-William
hal.structure.identifierBiologie des maladies cardiovasculaires = Biology of Cardiovascular Diseases
dc.contributor.authorDJABAROUTI, Sarah
hal.structure.identifierBiologie des maladies cardiovasculaires = Biology of Cardiovascular Diseases
dc.contributor.authorCOUFFINHAL, Thierry
hal.structure.identifierChimie et Biologie des Membranes et des Nanoobjets [CBMN]
dc.contributor.authorSCHMITTER, Jean-Marie
hal.structure.identifierChimie et Biologie des Membranes et des Nanoobjets [CBMN]
dc.contributor.authorBREILH, Dominique
dc.date.accessioned2020-07-09T14:16:45Z
dc.date.available2020-07-09T14:16:45Z
dc.date.issued2017-12-01
dc.identifier.issn1570-0232
dc.identifier.urihttps://oskar-bordeaux.fr/handle/20.500.12278/10297
dc.description.abstractEnThe quantification of monoclonal antibodies (mAbs) such as bevacizumab, a recombinant humanized immunoglobulin G1 (hIgG1), in biological fluids, is an essential prerequisite to any pharmacokinetic preclinical and clinical study. To date, reference techniques used to quantify mAbs rely on enzyme-linked immunosorbent assay (ELISA) lacking specificity. Furthermore, the commercially available ELISA kit to quantify bevacizumab in human plasma only assesses the free fraction of the drug. However, the conditions of storage and analysis of plasma samples could alter the physiological equilibrium between the free, bound and partially bound forms of bevacizumab and this could result in over- or underestimation of drug concentration. We developed a new assay for absolute quantification of total fraction of bevacizumab by liquid chromatography tandem mass spectrometry (LC-MS/MS) basing identification and quantification of bevacizumab on two specific peptides. In this report we compare our assay with two internal standard (IS) calibration approaches: one using a different human mAb (Trastuzumab) and the other using a stable isotope labeled specific peptide. After enrichment by affinity chromatography on protein A and concentration by ultrafiltration, human plasma samples were proteolyzed by trypsin. Linearity was established from 12.5 to 500 mu g/mL with an interday accuracy ranging from 101.7 to 110.6% and precision from 7.0% to 9.9%. This study demonstrates the importance of the choice of the IS in quantifying bevacizumab in human plasma and highlights the difficulty of reaching a reliable proteolysis with a sufficient recovery. We developed a reliable and cost-effective LC-MS/MS method to quantify total plasmatic fraction of bevacizumab in human plasma. Through our development we proposed a generic methodology easily transposable to quantify all IgG1 subclass very useful for clinical pharmacokinetics studies.
dc.title.enA new reliable, transposable and cost-effective assay for absolute quantification of total plasmatic bevacizumab by LC MS/MS in human plasma comparing two internal standard calibration approaches
dc.typeArticle de revue
dc.identifier.doi10.1016/j.jchromb.2017.10.042
dc.subject.halChimie/Matériaux
bordeaux.journalJournal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences
bordeaux.page43-53
bordeaux.volume1070
bordeaux.hal.laboratoriesInstitut de Chimie & de Biologie des Membranes & des Nano-objets (CBMN) - UMR 5248*
bordeaux.hal.laboratoriesInstitut de Chimie & de Biologie des Membranes & des Nano-objets (CBMN, UMR 5248)
bordeaux.institutionUniversité de Bordeaux
bordeaux.institutionBordeaux INP
bordeaux.COinSctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.jtitle=Journal%20of%20Chromatography%20B-Analytical%20Technologies%20in%20the%20Biomedical%20and%20Life%20Sciences&rft.date=2017-12-01&rft.volume=1070&rft.spage=43-53&rft.epage=43-53&rft.eissn=1570-0232&rft.issn=1570-0232&rft.au=LEGERON,%20Rachel&XUEREB,%20Fabien&CHAIGNEPAIN,%20Stephane&GADEAU,%20Alain-Pierre&CLAVEROL,%20Stephane&rft.genre=article


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