Fully validated LCMS bioanalysis of Bevacizumab in human plasma using nano-surface and molecular-orientation limited (nSMOL) proteolysis

Iwamoto, Noriko; Umino, Yukari; Aoki, Chikage; Yamane, Naoe; Hamada, Akinobu; Shimada, Takashi

doi:10.1016/j.dmpk.2015.11.004

Cited by 43 publications

(24 citation statements)

References 8 publications

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“…In conclusion, we have developed a new validated LC‐MS bioanalysis for the Fc‐fusion biopharmaceuticals Etanercept and Abatacept, using the nSMOL application like the case for several antibody drugs reported previously . To our knowledge, this is the first study to apply a direct quantitation of Etanercept and Abatacept in human serum using validated LC‐MS bioanalysis.…”

Section: Resultsmentioning

confidence: 90%

Application of nSMOL coupled with LC‐MS bioanalysis for monitoring the Fc‐fusion biopharmaceuticals Etanercept and Abatacept in human serum

Iwamoto

Yokoyama

Takanashi

et al. 2018

Pharmacology Res & Perspec

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The principle of nano‐surface and molecular‐orientation limited (nSMOL) proteolysis has a unique characteristic Fab‐selective proteolysis for antibody bioanalysis that is independent of a variety of monoclonal antibodies by the binding antibody Fc via Protein A/G in a pore with 100 nm diameter and modified trypsin immobilization on the surface of nanoparticles with 200 nm diameter. Since minimizing peptide complexity and protease contamination while maintaining antibody sequence specificity enables a rapid and broad development of optimized methods for liquid chromatography‐mass spectrometry (LC‐MS) bioanalysis, the application of regulatory LC‐MS for monitoring antibody biopharmaceuticals is expected. nSMOL is theoretically anticipated to be applicable for representative Fc‐fusion biopharmaceuticals, because Protein A/G‐binding site Fc exists on the C‐terminus, and its functional domain is available to orient and interact with the reaction solution. In this report, we describe the validated LC‐MS bioanalysis for monitoring Ethanercept and Abatacept using nSMOL technology. The quantitation range of Ethanercept in human serum was from 0.195 to 100 μg/mL using the signature peptide VFCTK (aa.43‐47), and that of Abatacept was from 0.391 to 100 μg/mL using the signature peptide MHVAQPAVVLASSR (aa.1‐14). Both proteins fulfilled the guideline criteria for low‐molecular‐weight drug compounds. The results indicate that the clinical and therapeutic monitoring for antibody and Fc‐fusion biopharmaceuticals are adequately applicable using nSMOL proteolysis coupled with LC‐MS bioanalysis.

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Section: Resultsmentioning

confidence: 90%

Application of nSMOL coupled with LC‐MS bioanalysis for monitoring the Fc‐fusion biopharmaceuticals Etanercept and Abatacept in human serum

Iwamoto

Yokoyama

Takanashi

et al. 2018

Pharmacology Res & Perspec

Self Cite

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“… 2 , 3 Over the last decade, liquid chromatography (LC)–mass spectrometry (MS)-based methods have thus arisen as an alternative for the quantification of therapeutic mAbs in plasma for PK evaluation in preclinical and clinical studies, 4 − 7 as for trastuzumab, 8 nivolumab, 9 , 10 infliximab, 11 , 12 or adalimumab 12 among others. 13 − 15 LC–MS-based assays enable to reach high specificity, broad dynamic ranges, and versatility, are usually faster to develop, can be easily standardized, and also offer the possibility to monitor several compounds simultaneously. 12 , 15 As a first development step in such assays, a specific investigation of the targeted antibody sequence and complementarity-determining regions (CDR) is necessary to guarantee the specificity of the analysis.…”

Section: Introductionmentioning

confidence: 99%

Development and Validation of an LC–MS-Based Quantification Assay for New Therapeutic Antibodies: Application to a Novel Therapy against Herpes Simplex Virus

et al. 2020

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Multiple therapeutic monoclonal antibodies (mAbs) are currently under development or in (pre)clinical study phases to reach regulatory approval. Among these, a new mAb against herpes simplex virus, HDIT101, was recently tested in healthy volunteers during a phase I clinical trial (first-in-human, dose escalation). In the frame of the pharmacokinetic evaluation of this new therapy, a mass spectrometric (MS)-based method was developed for the quantification of HDIT101 in human plasma using liquid chromatography coupled to tandem mass spectrometry. In this work, we describe the development of this bioanalytical assay using the quantification of a HDIT101 surrogate peptide, the assay validation procedure according to the FDA guidelines within the calibration range from 20 to 5000 μg/mL, and its application to plasma samples from the first-in-human clinical trial. This work presents a generic workflow for the development of MS-based quantification assays of new therapeutic antibodies that allows reaching high immunopurification recovery (>98% for HDIT101 over the full calibration range with a precision of 6.9% CV). Surrogate peptide and stable isotopically labeled internal standard were stable, and batch-to-batch accuracies and precisions at the four quality standard levels ranged between −2 and 5% bias and 8 and 11% CV, respectively.

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“…Therefore, the use of immunoaffinity purification may be an effective pretreatment approach even with a tryptic digestion–LC/MS/MS method. In this direction, various affinity ligand molecules such as anti-drug antibody [9], protein A [10], and protein G [11,12] have been reported. The recent improvement in the performance of high-resolution mass spectrometers has allowed quantification of therapeutic mAbs treated with affinity purification as intact or Fab-fragmented forms without tryptic digestion [13,14,15].…”

Section: Introductionmentioning

confidence: 99%

Anti-Idiotype DNA Aptamer Affinity Purification–High-Temperature Reversed-Phase Liquid Chromatography: A Simple, Accurate, and Selective Bioanalysis of Bevacizumab

et al. 2019

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This study presents a simple, accurate, and selective bioanalytical method of bevacizumab detection from plasma samples based on aptamer affinity purification–high-temperature reversed-phased liquid chromatography (HT-RPLC) with fluorescence detection. Bevacizumab in plasma samples was purified using magnetic beads immobilized with an anti-idiotype DNA aptamer for bevacizumab. The purified bevacizumab was separated with HT-RPLC and detected with its native fluorescence. Using aptamer affinity beads, bevacizumab was selectively purified and detected as a single peak in the chromatogram. HT-RPLC achieved good separation for bevacizumab with a sharp peak within 10 min. The calibration curves of the two monoclonal antibodies ranged from 1 to 50 μg/mL and showed good correlation coefficients (r2 > 0.999). The limit of detection (LOD) and lower limit of quantification (LLOQ) values for bevacizumab were 0.15 and 0.51 μg/mL, respectively. The proposed method was successfully applied to the bioanalysis of the plasma samples obtained from the patients with lung cancer and may be extended to plan optimal therapeutic programs and for the evaluation of biological equivalencies in the development of biosimilars.

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Fully validated LCMS bioanalysis of Bevacizumab in human plasma using nano-surface and molecular-orientation limited (nSMOL) proteolysis

Cited by 43 publications

References 8 publications

Application of nSMOL coupled with LC‐MS bioanalysis for monitoring the Fc‐fusion biopharmaceuticals Etanercept and Abatacept in human serum

Application of nSMOL coupled with LC‐MS bioanalysis for monitoring the Fc‐fusion biopharmaceuticals Etanercept and Abatacept in human serum

Development and Validation of an LC–MS-Based Quantification Assay for New Therapeutic Antibodies: Application to a Novel Therapy against Herpes Simplex Virus

Anti-Idiotype DNA Aptamer Affinity Purification–High-Temperature Reversed-Phase Liquid Chromatography: A Simple, Accurate, and Selective Bioanalysis of Bevacizumab

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