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Infliximab quantitation in human plasma by liquid chromatography-tandem mass spectrometry: towards a standardization of the methods?

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Abstract

Infliximab (IFX) is a chimeric monoclonal antibody targeting tumor necrosis factor-alpha. It is currently approved for the treatment of certain rheumatic diseases or inflammatory bowel diseases. Clinical studies have suggested that monitoring IFX concentrations could improve treatment response. However, in most studies, IFX was quantified using ELISA assays, the resulting discrepancies of which raised concerns about their reliability. Here, we describe the development and validation of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for IFX quantification in human plasma. Full-length stable-isotope-labeled antibody (SIL-IFX) was added to plasma samples as internal standard. Samples were then prepared using Mass Spectrometry Immuno Assay (MSIA™) followed by trypsin digestion and submitted to multiple reaction monitoring (MRM) for quantification of IFX. The chromatographic run lasted 13 min. The range of quantification was 1 to 26 mg/L. For two internal quality controls spiked with 6 and 12 mg/L of IFX, the method was reproducible (coefficients of variation (CV%): 12.7 and 2.1), repeatable (intra-day CV%: 5.5 and 5.0), and accurate (inter-day and intra-day deviations from nominal values: +6.4 to +3.7 % and 5.5 to 9.2 %, respectively). There was no cross - contamination effect. Samples from 45 patients treated with IFX were retrospectively analyzed by LC-MS/MS and results were compared to those obtained with an in-house ELISA assay and the commercial Lisa Tracker® method. Good agreement was found between LC-MS/MS and in-house ELISA (mean underestimation of 13 % for in-house ELISA), but a significant bias was found with commercial ELISA (mean underestimation of 136 % for commercial ELISA). This method will make it possible to standardize IFX quantification between laboratories.

Interassay comparison of the three methods: LC-MS/MS vs inhouse ELISA assay or vs Lisa Tracker® ELISA assays, Passing & Bablok (a) and Bland & Altman (b) for the comparison of LC-MS/MS vs in-house ELISA assay; Passing & Bablok (c); Bland & Altman (d) for the comparison LC-MS/MS vs Lisa Tracker® ELISA assay

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Acknowledgments

We thank Ravindra Chaudhari from Thermo-Fisher for advice and technical assistance with the MSIA procedure. We are also indebted to Karine Scalabrino for excellent relevant technical assistance.

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Authors and Affiliations

  1. Grenoble-Alpes University Hospital, Hospital Albert Michalon, Laboratory of Pharmacology and Toxicology, CS10217, 38043, Grenoble, France

    Jean-Francois Jourdil, Elodie Gautier-Veyret, Bruno Bonaz, Julia Tonini & Françoise Stanke-Labesque

  2. PROMISE Advanced Proteomics, 7 Parvis Louis Néel, 38040, Grenoble, France

    Dorothée Lebert & Guillaume Picard

  3. University Grenoble Alpes, 38041, Grenoble, France

    Elodie Gautier-Veyret, Bruno Bonaz & Françoise Stanke-Labesque

  4. INSERM U1042, HP2, 38041, Grenoble, France

    Elodie Gautier-Veyret

  5. Department of Clinical and Biological Pharmacology and Pharmacovigilance, Pharmacoepidemiology and Drug Information Center, University Hospital Rennes, 35000, Rennes, France

    Florian Lemaitre

  6. IINSERM, U1216, 38000, Grenoble, France

    Bruno Bonaz

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  1. Jean-Francois Jourdil
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  2. Dorothée Lebert
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Correspondence to Jean-Francois Jourdil.

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This study has been approved by the Grenoble University Hospital ethics committee IRB6705 and has been performed in accordance with the ethical standards.

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The authors declare that they have no conflict of interest.

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Jourdil, JF., Lebert, D., Gautier-Veyret, E. et al. Infliximab quantitation in human plasma by liquid chromatography-tandem mass spectrometry: towards a standardization of the methods?. Anal Bioanal Chem 409, 1195–1205 (2017). https://doi.org/10.1007/s00216-016-0045-4

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  • DOI: https://doi.org/10.1007/s00216-016-0045-4

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