Quantitation and modeling of post-translational modifications in a therapeutic monoclonal antibody from single- and multiple-dose monkey pharmacokinetic studies using mass spectrometry

Xu, Xiaobin; Huang, Yi-Hsiang; Pan, Hao; Molden, Rosalynn C.; Qiu, Haibo; Daly, Thomas J.; Li, Ning

doi:10.1371/journal.pone.0223899

Cited by 15 publications

(13 citation statements)

References 28 publications

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“…LBAs may utilize different combinations of generic anti-human IgG, therapeutic target, anti-idiotype and non-neutralizing antiidiotype reagents for capture or detection [1][2][3][4]. Over time, MS-based workflows progressed to enable quantitation of protein therapeutics with a digestion-based, surrogate peptide approach [5][6][7][8], where a single peptide is used to infer protein concentration and analysis is typically carried out on a triple quadrupole (QQQ) mass spectrometer [9]. For biotherapeutic peptides and small proteins, several examples have demonstrated intact mass quantification on triple-QQQ and high-resolution MS (HRMS) systems [10][11][12][13].…”

mentioning

confidence: 99%

Intact mAb LC–MS for Drug Concentration from Pre-Clinical Studies: Bioanalytical Method Performance and in-Life Samples

et al. 2020

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Background: Antibody biotherapeutic measurement from pharmacokinetic studies has not been traditionally based on intact molecular mass as is the case for small molecules. However, recent advancements in protein capture and mass spectrometer technology have enabled intact mass detection and quantitation for dosed biotherapeutics. A bioanalytical method validation is part of the regulatory requirement for sample analysis to determine drug concentration from in-life study samples. Results/methodology: Here, an intact protein LC–MS assay is subjected to mock bioanalytical method validation, and unknown samples are compared between intact protein LC–MS and established bioanalytical assay formats: Ligand-binding assay and peptide LC–MS/MS. Discussion/conclusion: Results are presented from the intact and traditional bioanalytical method evaluations, where the in-life sample concentrations were comparable across method types with associated data analyses presented. Furthermore, for intact protein LC–MS, modification monitoring and evaluation of data processing parameters is demonstrated.

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confidence: 99%

Intact mAb LC–MS for Drug Concentration from Pre-Clinical Studies: Bioanalytical Method Performance and in-Life Samples

et al. 2020

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“…CQAs may impact pharmacokinetics, binding properties, or immunogenicity, as they are product and application specific, their identification needs to be assessed by specific in vitro studies. Further in vivo studies are also required to confirm which attributes are indeed critical and should be included in the product specification criteria [ 16 , 17 ].…”

Section: Introductionmentioning

confidence: 99%

Multi attribute method implementation using a High Resolution Mass Spectrometry platform: From sample preparation to batch analysis

et al. 2022

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Quality control of biopharmaceuticals such as monoclonal antibodies (mAbs) has been evolving and becoming more challenging as the requirements of the regulatory agencies increase due to the demanding complexity of products under evaluation. Mass Spectrometry (MS)-based methods such as the multi-attribute method (MAM) are being explored to achieve a deeper understanding of the attributes critical for the safety, efficacy, and quality of these products. MAM uses high mass accuracy/high-resolution MS data that enables the direct and simultaneous monitoring of relevant product quality attributes (PQAs, in particular, chemical modifications) in a single workflow, replacing several orthogonal methods, reducing time and costs associated with these assays. Here we describe a MAM implementation process using a QTOF high resolution platform. Method implementation was accomplished using NIST (National Institute for Standards and Technology) mAb reference material and an in-process mAb sample. PQAs as glycosylation profiles, methionine oxidation, tryptophan dioxidation, asparagine deamidation, pyro-Glu at N-terminal and glycation were monitored. Focusing on applications that require batch analysis and high-throughput, sample preparation and LC-MS parameters troubleshooting are discussed. This MAM workflow was successfully explored as reference analytical tool for comprehensive characterization of a downstream processing (DSP) polishing platform and for a comparability study following technology transfer between different laboratories.

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“…With the rapid development of new instrumentation and methods, liquid chromatography-mass spectrometry (LC-MS) has emerged as a critical tool for the characterization of biologicals (13)(14)(15). Many efforts have been made to adopt the LC-MS-based approach as a supplementary tool for ADA analysis (16).…”

Section: Introductionmentioning

confidence: 99%

Isotyping and Semi-Quantitation of Monkey Anti-Drug Antibodies by Immunocapture Liquid Chromatography-Mass Spectrometry

Huang

Chen

et al. 2021

AAPS J

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There is an urgent demand to develop new technologies to characterize immunogenicity to biotherapeutics. Here, we developed an immunocapture LC-MS assay to isotype and semi-quantify monkey anti-drug antibodies (ADAs) to fully human monoclonal antibody (mAb) drugs. ADAs were isolated from serum samples using an immunocapture step with the Fab of the full-length mAb cross-linked to magnetic beads to minimize matrix interference. A positive monoclonal antibody control against the human immunoglobulin kappa light chain was used as a calibration standard for ADA quantitation. The final LC-MS method contains 17 multiple reaction monitoring (MRM) transitions and an optimized 15-min LC method. The results suggested that IgG1 was the most abundant isotype in ADA-positive samples. IgG2 and IgG4 were identified at lower levels, whereas IgG3 and IgA levels were only observed at very minor levels. In addition, levels of total ADA measured by the LC-MS assay were comparable to results obtained using a traditional ligand binding assay (LBA). The LC-MS ADA assay enabled rapid immunogenicity assessment with additional isotype information that LBAs cannot provide.

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Quantitation and modeling of post-translational modifications in a therapeutic monoclonal antibody from single- and multiple-dose monkey pharmacokinetic studies using mass spectrometry

Cited by 15 publications

References 28 publications

Intact mAb LC–MS for Drug Concentration from Pre-Clinical Studies: Bioanalytical Method Performance and in-Life Samples

Intact mAb LC–MS for Drug Concentration from Pre-Clinical Studies: Bioanalytical Method Performance and in-Life Samples

Multi attribute method implementation using a High Resolution Mass Spectrometry platform: From sample preparation to batch analysis

Isotyping and Semi-Quantitation of Monkey Anti-Drug Antibodies by Immunocapture Liquid Chromatography-Mass Spectrometry

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