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Validated LC/MS Bioanalysis of Rituximab CDR Peptides Using Nano-surface and Molecular-Orientation Limited (nSMOL) Proteolysis

Iwamoto, Noriko ; Takanashi, Megumi ; Hamada, Akinobu ; Shimada, Takashi

Biological and Pharmaceutical Bulletin, 2016/07/01, Vol.39(7), pp.1187-1194 [Periódico revisado por pares]

Japan: The Pharmaceutical Society of Japan

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  • Título:
    Validated LC/MS Bioanalysis of Rituximab CDR Peptides Using Nano-surface and Molecular-Orientation Limited (nSMOL) Proteolysis
  • Autor: Iwamoto, Noriko ; Takanashi, Megumi ; Hamada, Akinobu ; Shimada, Takashi
  • Assuntos: antibody drug ; bioanalysis ; Chromatography, Liquid ; clinical pharmacokinetics ; Complementarity Determining Regions ; Humans ; LC/MS ; Limit of Detection ; nano-surface and molecular-orientation limited (nSMOL) proteolysis ; Peptides - blood ; Peptides - chemistry ; Proteolysis ; Reproducibility of Results ; Rituximab ; Rituximab - blood ; Rituximab - chemistry ; Tandem Mass Spectrometry
  • É parte de: Biological and Pharmaceutical Bulletin, 2016/07/01, Vol.39(7), pp.1187-1194
  • Notas: ObjectType-Article-1
    SourceType-Scholarly Journals-1
    ObjectType-Feature-2
    content type line 23
  • Descrição: Presently, monoclonal antibodies (mAbs) therapeutics have big global sales and are starting to receive competition from biosimilars. We previously reported that the nano-surface and molecular-orientation limited (nSMOL) proteolysis which is optimal method for bioanalysis of antibody drugs in plasma. The nSMOL is a Fab-selective limited proteolysis, which utilize the difference of protease nanoparticle diameter (200 nm) and antibody resin pore diameter (100 nm). In this report, we have demonstrated that the full validation for chimeric antibody Rituximab bioanalysis in human plasma using nSMOL proteolysis. The immunoglobulin fraction was collected using Protein A resin from plasma, which was then followed by the nSMOL proteolysis using the FG nanoparticle-immobilized trypsin under a nondenaturing condition at 50°C for 6 h. After removal of resin and nanoparticles, Rituximab signature peptides (GLEWIGAIYPGNGDTSYNQK, ASGYTFTSYNMHWVK, and FSGSGSGTSYSLTISR) including complementarity-determining region (CDR) and internal standard P14R were simultaneously quantified by multiple reaction monitoring (MRM). This quantification of Rituximab using nSMOL proteolysis showed lower limit of quantification (LLOQ) of 0.586 µg/mL and linearity of 0.586 to 300 µg/mL. The intra- and inter-assay precision of LLOQ, low quality control (LQC), middle quality control (MQC), and high quality control (HQC) was 5.45–12.9% and 11.8, 5.77–8.84% and 9.22, 2.58–6.39 and 6.48%, and 2.69–7.29 and 4.77%, respectively. These results indicate that nSMOL can be applied to clinical pharmacokinetics study of Rituximab, based on the precise analysis.
  • Editor: Japan: The Pharmaceutical Society of Japan
  • Idioma: Inglês;Japonês
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  • Realização: ABCD-USP USP   Logos de Redes Sociais: Freepik

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