Determination of plasma concentration of Belimumab by LC-MS/MS: Method development, validation, and clinical application

Clémence Marin; Gaëlle Noé; Dimitri Schlemmer; Quentin Beaulieu; Pascal Robidou; Bochra Mansour; Christophe Hirtz; Jérôme Vialaret; Marie Antignac; Quentin Moyon; Neila Benameur; Zahir Amoura; Noël Zahr

doi:10.1016/j.jpba.2023.115730

Determination of plasma concentration of Belimumab by LC-MS/MS: Method development, validation, and clinical application

J Pharm Biomed Anal. 2023 Nov 30:236:115730. doi: 10.1016/j.jpba.2023.115730. Epub 2023 Sep 17.

Authors

Affiliations

¹ AP-HP Sorbonne Université, Pitié-Salpêtrière Hospital, Department of Pharmacology, CIC-1901, Pharmacokinetics and Therapeutic Drug Monitoring Unit, UMR-S 1166, F-75013 Paris, France.
² AP-HP. Sorbonne Université, Laboratoire de suivi thérapeutique pharmacologique spécialisé, F-75013 Paris, France.
³ IRMB-PPC, INM, Montpellier University Hospital, INSERM, CNRS, University of Montpellier, 34295 Montpellier, France.
⁴ Department of Pharmacy, PITIE-SALPETRIERE Hospital, AP-HP Sorbonne université, F-75013 Paris, France.
⁵ Sorbonne Université, Assistance Publique-Hôpitaux de Paris (AP-HP), Groupement Hospitalier Pitié-Salpêtrière, Centre de Référence pour le Lupus, le Syndrome des anti-phospholipides et autres maladies auto-immunes rares, Service de Médecine Interne 2, Paris, France.
⁶ AP-HP Sorbonne Université, Pitié-Salpêtrière Hospital, Department of Pharmacology, CIC-1901, Pharmacokinetics and Therapeutic Drug Monitoring Unit, UMR-S 1166, F-75013 Paris, France; AP-HP. Sorbonne Université, Laboratoire de suivi thérapeutique pharmacologique spécialisé, F-75013 Paris, France. Electronic address: noel.zahr@aphp.fr.

PMID: 37734255
DOI: 10.1016/j.jpba.2023.115730

Abstract

Introduction: Belimumab is a monoclonal antibody against B cell activating factor (BLyS). This monoclonal antibody (mAb) has been shown to be effective in reducing disease activity in patients with systemic lupus erythematosus (SLE). Belimumab is available in two forms as a lyophilized powder for intravenous (IV) use, or single-dose syringe for subcutaneous (SC) use. The aim of this study was to develop and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for quantitation of belimumab in human serum.

Material and methods: All analyses relied on nano-surface and molecular-orientation limited (nSMOL) proteolysis coupled with LC-MS/MS. Quantifications was performed in multiple reactions monitoring (MRM) mode, and electrospray ionization was conducted in positive mode.

Results: Belimumab was quantified with signature peptide QAPGQGLEWMGGIPFGTAK and normalized using P14R. The total run time per assay was 10 min. Linearity was measured from 5 to 800 μg/mL (r² > 0.995). Accuracy and precision based on three quality control levels range from 11.2 - 9.51 % and 1.24 - 13.12 % respectively. The carryover was less than 7 %. In all, 87 patient samples were processed (65, IV; 22, SC). Mean concentration of belimumab was significantly higher for SC (93.0 ± 74.0 µg/mL) than for IV (67.4 ± 38.9 µg/mL) administration.

Conclusion: We have developed the first method of belimumab quantification combining LC-MS/MS and nSMOL proteolysis. It can be used for future clinical pharmacokinetic studies of belimumab and for investigating the relationship between belimumab concentration, efficacy, and toxicity in SLE patients.

Keywords: Belimumab; Lupus; Pharmacokinetic; Therapeutic Drug Monitoring (TDM).