Multiplexed monitoring of therapeutic antibodies for inflammatory diseases using Fab-selective proteolysis nSMOL coupled with LC-MS

Iwamoto, Noriko; Takanashi, Megumi; Yokoyama, Kotoko; Yonezawa, Atsushi; Denda, Masaya; Hashimoto, Motomu; Tanaka, Masao; Ito, Hiromu; Matsuura, Minoru; Yamamoto, Shuji; Honzawa, Yusuke; Matsubara, Kazuo; Shimada, Takashi

doi:10.1016/j.jim.2019.06.014

Cited by 22 publications

(10 citation statements)

References 31 publications

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“…Further, the best LLOQ was 0.06 μg/ml in the trastuzumab assay 50 . The nSMOL assay has an excellent linear quantitative range: the LLOQ of the nSMOL assay is typically lower than 1 μg/mL and it can measure mAbs at up to 100-300 μg/ml 51 . This feature curtails errors introduced through sample dilution and during the process of extrapolation of mAb concentration from the standard curve, while such errors are difficult to mitigate in ELISA (e.g., a linear standard curve is typically accompanied by narrower dynamic range whereas wider dynamic range ends up with a sigmoidal standard curve).…”

Section: Discussionmentioning

confidence: 99%

“…These patients can be treated with other mAb targeting IL-12/23p40, C5, or IL-6R at a medical center for autoimmunity and inflammation. Therefore, we developed multiplexed nSMOL assay to simultaneously detect 9 mAbs such as infliximab, adalimumab, ustekinumab, golimumab, eculizumab, etanercept, abatacept, tocilizumab, and mepolizumab in a single run and successfully detected mAbs from patient samples 41,51 . Others have also reported simultaneous detection of 2-7 mAbs 19,41,55-58 , further proving the feasibility of multiplexed detection of mAbs by LC-MS.…”

Section: Discussionmentioning

confidence: 99%

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A rapid and universal liquid chromatograph-mass spectrometry-based platform, refmAb-Q nSMOL, for monitoring monoclonal antibody therapeutics

Iwamoto

Koguchi

Yokoyama

et al. 2022

Preprint

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Accurate quantitation of antibody is critical for development of monoclonal antibody therapeutics (mAbs). Therapeutic drug monitoring has been applied to measure levels of mAbs in clinics for dose adjustment for autoimmune disease. Trough levels of mAbs can be a biomarker for cancer immunotherapy. Thus, the deployment of a rapid and universal platform for mAb monitoring may benefit processes ranging from drug development to clinical practice for a wide spectrum of diseases. However, mAb monitoring often requires development and conduct of an individual ligand binding assay such as ELISA, which is impractical to scale. We streamlined quantitation of antibody therapeutics by a nano-surface and molecular-orientation limited (nSMOL) proteolysis assay using LC-MS with a universal reference antibody (refmAb-Q), for accurate multiplexed quantitation of unique signature peptides derived from mAbs. This innovative refmAb-Q nSMOL platform may provide a practical solution for quantitating an ever-increasing number of mAbs from developmental to clinical use settings.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

A rapid and universal liquid chromatograph-mass spectrometry-based platform, refmAb-Q nSMOL, for monitoring monoclonal antibody therapeutics

Iwamoto

Koguchi

Yokoyama

et al. 2022

Preprint

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“…Blood samples for measuring trough serum IFX levels were collected immediately before a new infusion. Serum IFX levels were measured using an LCMS-8060 quadrupole mass spectrometer (SHIMADZU, Kyoto, Japan), as previously reported, with some modifications [ 24 – 26 ]. Briefly, to obtain the peptides from the fragment antigen-binding (Fab) region of immunoglobulin G, serum samples were pretreated using the nSMOL ™ Antibody BA Kit (SHIMADZU, Kyoto, Japan) according to the provided protocol.…”

Section: Methodsmentioning

confidence: 99%

Potential application of measuring serum infliximab levels in rheumatoid arthritis management: A retrospective study based on KURAMA cohort data

et al. 2021

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Infliximab (IFX) therapy has considerably improved the treatment of rheumatoid arthritis (RA). However, some patients still do not respond adequately to IFX therapy, or the efficacy of the treatment diminishes over time. Although previous studies have reported a relationship between serum IFX levels and therapeutic efficacy, the potential applications of IFX therapeutic drug monitoring (TDM) in clinical practice remain unclear. The purpose of this study was to investigate the potential applications of IFX TDM by analyzing a Japanese cohort database. Data were collected retrospectively from the Kyoto University Rheumatoid Arthritis Management Alliance cohort between January 1, 2011, and December 31, 2018. Serum IFX levels were measured using a liquid chromatography-tandem mass spectrometer. Out of the 311 RA patients that used IFX, 41 were eligible for the analysis. Serum IFX levels were significantly higher in responders than in non-responders. An optimal cut-off value was determined to be 0.32 μg/mL based on a receiver operating characteristic curve. At the IFX measurement point, a better therapeutic response was observed in the high IFX group (n = 32) than in the low IFX group (n = 9). Conversely, at the maximum effect point, when DAS28-ESR was the lowest between IFX introduction and measurement points, there were no differences in responder proportions between the low and high IFX groups. IFX primary ineffectiveness could be avoided with appropriate dose escalation without blood concentration measurement in clinical practice. In conclusion, IFX TDM could facilitate the identification of secondary non-responders and in turn, proper IFX use.

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“…Blood samples for measuring trough serum IFX levels were collected immediately before a new infusion. Serum IFX levels were measured using an LCMS-8060 quadrupole mass spectrometer (SHIMADZU, Kyoto, Japan), as previously reported, with some modifications [24][25][26]. Briefly, to obtain the peptides from the fragment antigen-binding (Fab) region of immunoglobulin G, serum samples were pretreated using the nSMOL TM Antibody BA Kit (SHIMADZU, Kyoto, Japan) according to the provided protocol.…”

Section: Measurement Of Serum Ifx Levelsmentioning

confidence: 99%

Potential Application of Measuring Serum Infliximab Levels in Rheumatoid Arthritis Management: A Retrospective Study based on KURAMA Cohort Data

Nakae

Masui

Yonezawa

et al. 2021

Preprint

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Background Infliximab (IFX) therapy has considerably improved the treatment of rheumatoid arthritis (RA). On the other hand, in some patients, the efficacy of IFX therapy is not adequate, or gradually diminishes with the lapse of the treatment. Although previous studies have reported a positive relationship between serum IFX levels and therapeutic efficacy, the potential application of IFX therapeutic drug monitoring (TDM) in clinical practice remains unclear. The purpose of this study was to investigate the potential applications of IFX TDM by analyzing a Japanese cohort database. Methods Data were collected retrospectively from the Kyoto University Rheumatoid Arthritis Management Alliance, KURAMA,cohort between January 1, 2011, and December 31, 2018. Serum IFX levels were measured using liquid chromatography-tandem mass spectrometry. Results Out of the 311 RA patients who received IFX therapy, 41 were eligible for analysis. Serum IFX levels were significantly higher in responders than in non-responders. An optimal cut-off value was determined to be 0.4 µg/mL based on a receiver operating characteristic curve. At the IFX measurement point, a better therapeutic response was observed in the High-IFX group (n = 31) than in the Low-IFX group (n = 10). Conversely, at the maximum effect point, when DAS28-ESR (the 28 joint disease activity score incorporating erythrocyte sedimentation rate) was the lowest between IFX introduction and measurement points, there were no differences in responder proportions between the Low- and High-IFX groups. Conclusions In clinical practice, IFX primary ineffectiveness could be avoided with appropriate dose escalation without blood concentration measurement. However, IFX TDM could facilitate the identification of secondary non-responders, and in turn, proper IFX use.

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Multiplexed monitoring of therapeutic antibodies for inflammatory diseases using Fab-selective proteolysis nSMOL coupled with LC-MS

Cited by 22 publications

References 31 publications

A rapid and universal liquid chromatograph-mass spectrometry-based platform, refmAb-Q nSMOL, for monitoring monoclonal antibody therapeutics

A rapid and universal liquid chromatograph-mass spectrometry-based platform, refmAb-Q nSMOL, for monitoring monoclonal antibody therapeutics

Potential application of measuring serum infliximab levels in rheumatoid arthritis management: A retrospective study based on KURAMA cohort data

Potential Application of Measuring Serum Infliximab Levels in Rheumatoid Arthritis Management: A Retrospective Study based on KURAMA Cohort Data

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