Ocrelizumab quantitation by liquid chromatography-tandem mass spectrometry.
J Mass Spectrom Adv Clin Lab
; 25: 53-60, 2022 Aug.
Article
em En
| MEDLINE
| ID: mdl-35910410
ABSTRACT
Introduction:
Ocrelizumab is a monoclonal anti-CD20 antibody approved for the treatment of multiple sclerosis (MS). The clinical value of therapeutic drug monitoring (TDM) for this antibody in treatment of MS is unknown, and an adequately specific and precise quantitation method for ocrelizumab in patient serum could facilitate investigation. Liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based quantitation methods have been shown to have higher analytic specificity and precision than enzyme-linked immunosorbent assays.Objectives:
To establish and validate an LC-MS/MS-based quantitation method for ocrelizumab.Methods:
We present an LC-MS/MS-based quantitation method using immunocapture purification followed by trypsinization and analysis by a triple quadrupole mass analyzer obtaining results within the same day.Results:
We found that the ocrelizumab peptide GLEWVGAIYPGNGDTSYNQK (Q1/Q3 Quantifier ion 723.683+/590.77 y112+ Qualifier ion 723.683+/672.30 y122+) can be used for quantitation and thereby developed a method for quantifying ocrelizumab in human serum with a quantitation range of 1.56 to 200 µg/mL. The method was validated in accordance with EMA requirements in terms of selectivity, carry-over, lower limit of quantitation, calibration curve, accuracy, precision and matrix effect. Ocrelizumab serum concentrations were measured in three MS patients treated with ocrelizumab, immediately before and after ocrelizumab infusion, with additional sampling after 2, 4, 8 and 12 weeks. Measured serum concentrations of ocrelizumab showed expected values for both Cmax and drug half-life over the sampled time period.Conclusion:
We have established a reliable quantitation method for serum ocrelizumab that can be applied in clinical studies, facilitating the evaluation of ocrelizumab TDM in MS.
EMA, European Medicines Agency; IDA, Information dependent acqusition; IgG1, immunoglobulin G1; LC-MS/MS; LC-MS/MS, Liquid chromatography-tandem mass spectrometry; MS, Multiple sclerosis; MWCO, molecular weight cut-off; Multiple sclerosis; OVERLORD-MS, Ocrelizumab vErsus Rituximab Off-Label at the Onset of Relapsing MS; Ocrelizumab; QTOF, quadrupole time-of-flight; QTRAP, quadrupole ion trap; ROS-MS, Rituximab and Ocrelizumab in Serum with Multiple Sclerosis study; TDM, therapeutic drug monitoring; Therapeutic drug monitoring; Therapeutic monoclonal antibodies; UHPLC, ultra high performance liquid chromatography; t-mAb, therapeutic monoclonal antibody
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MEDLINE
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En
Ano de publicação:
2022
Tipo de documento:
Article