Using Mass Spectrometry to Quantify Rituximab and Perform Individualized Immunoglobulin Phenotyping in ANCA-Associated Vasculitis

John R. Mills, Divi Cornec, Surendra Dasari, Paula M. Ladwig, Amber M. Hummel, Melissa Cheu, David L. Murray, Maria A. Willrich, Melissa R. Snyder, Gary S. Hoffman, Cees G M Kallenberg, Carol A. Langford, Peter A. Merkel, Paul A. Monach, Philip Seo, Robert F. Spiera, E. William St Clair, John H. Stone, Ulrich Specks, David R. Barnidge

Research output: Contribution to journalArticle

Abstract

Therapeutic monoclonal immunoglobulins (mAbs) are used to treat patients with a wide range of disorders including autoimmune diseases. As pharmaceutical companies bring more fully humanized therapeutic mAb drugs to the healthcare market analytical platforms that perform therapeutic drug monitoring (TDM) without relying on mAb specific reagents will be needed. In this study we demonstrate that liquid-chromatography-mass spectrometry (LC-MS) can be used to perform TDM of mAbs in the same manner as smaller nonbiologic drugs. The assay uses commercially available reagents combined with heavy and light chain disulfide bond reduction followed by light chain analysis by microflow-LC-electrospray ionization-quadrupole-time-of-flight mass spectrometry (ESI-Q-TOF MS). Quantification is performed using the peak areas from multiply charged mAb light chain ions using an in-house developed software package developed for TDM of mAbs. The data presented here demonstrate the ability of an LC-MS assay to quantify a therapeutic mAb in a large cohort of patients in a clinical trial. The ability to quantify any mAb in serum via the reduced light chain without the need for reagents specific for each mAb demonstrates the unique capabilities of LC-MS. This fact, coupled with the ability to phenotype a patient's polyclonal repertoire in the same analysis further shows the potential of this approach to mAb analysis.

Original languageEnglish (US)
Pages (from-to)6317-6325
Number of pages9
JournalAnalytical Chemistry
Volume88
Issue number12
DOIs
StatePublished - Jun 21 2016

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Antineutrophil Cytoplasmic Antibodies
Mass spectrometry
Immunoglobulins
Liquid chromatography
Pharmaceutical Preparations
Monitoring
Assays
Electrospray ionization
Software packages
Disulfides
Rituximab
Ions
Industry

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Mills, J. R., Cornec, D., Dasari, S., Ladwig, P. M., Hummel, A. M., Cheu, M., ... Barnidge, D. R. (2016). Using Mass Spectrometry to Quantify Rituximab and Perform Individualized Immunoglobulin Phenotyping in ANCA-Associated Vasculitis. Analytical Chemistry, 88(12), 6317-6325. https://doi.org/10.1021/acs.analchem.6b00544

Using Mass Spectrometry to Quantify Rituximab and Perform Individualized Immunoglobulin Phenotyping in ANCA-Associated Vasculitis. / Mills, John R.; Cornec, Divi; Dasari, Surendra; Ladwig, Paula M.; Hummel, Amber M.; Cheu, Melissa; Murray, David L.; Willrich, Maria A.; Snyder, Melissa R.; Hoffman, Gary S.; Kallenberg, Cees G M; Langford, Carol A.; Merkel, Peter A.; Monach, Paul A.; Seo, Philip; Spiera, Robert F.; St Clair, E. William; Stone, John H.; Specks, Ulrich; Barnidge, David R.

In: Analytical Chemistry, Vol. 88, No. 12, 21.06.2016, p. 6317-6325.

Research output: Contribution to journalArticle

Mills, JR, Cornec, D, Dasari, S, Ladwig, PM, Hummel, AM, Cheu, M, Murray, DL, Willrich, MA, Snyder, MR, Hoffman, GS, Kallenberg, CGM, Langford, CA, Merkel, PA, Monach, PA, Seo, P, Spiera, RF, St Clair, EW, Stone, JH, Specks, U & Barnidge, DR 2016, 'Using Mass Spectrometry to Quantify Rituximab and Perform Individualized Immunoglobulin Phenotyping in ANCA-Associated Vasculitis', Analytical Chemistry, vol. 88, no. 12, pp. 6317-6325. https://doi.org/10.1021/acs.analchem.6b00544
Mills, John R. ; Cornec, Divi ; Dasari, Surendra ; Ladwig, Paula M. ; Hummel, Amber M. ; Cheu, Melissa ; Murray, David L. ; Willrich, Maria A. ; Snyder, Melissa R. ; Hoffman, Gary S. ; Kallenberg, Cees G M ; Langford, Carol A. ; Merkel, Peter A. ; Monach, Paul A. ; Seo, Philip ; Spiera, Robert F. ; St Clair, E. William ; Stone, John H. ; Specks, Ulrich ; Barnidge, David R. / Using Mass Spectrometry to Quantify Rituximab and Perform Individualized Immunoglobulin Phenotyping in ANCA-Associated Vasculitis. In: Analytical Chemistry. 2016 ; Vol. 88, No. 12. pp. 6317-6325.
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