The AAPS Journal

, 21:101 | Cite as

Comparison of Bevacizumab Quantification Results in Plasma of Non-small Cell Lung Cancer Patients Using Bioanalytical Techniques Between LC-MS/MS, ELISA, and Microfluidic-based Immunoassay

  • Noriko Iwamoto
  • Megumi Takanashi
  • Takashi ShimadaEmail author
  • Jiichiro Sasaki
  • Akinobu HamadaEmail author
Research Article


The development of analytical techniques to study therapeutic monoclonal antibodies is expected to be useful for pharmacokinetic analysis and for the development of therapeutic indexes to determine dosage standards. To date, the blood concentration of antibody drugs has been analyzed by the enzyme-linked immunosorbent assay (ELISA). However, with the development of mass spectrometry and microfluidization technologies, the assay implication is drastically changing. We have developed an analytical validation method for many monoclonal antibodies and Fc-fusion proteins using Fab-selective proteolysis nSMOL coupled with liquid chromatography-mass spectrometry (LC-MS/MS). However, the correlation between the analyzed data characterization and the referable value from individual measurement techniques has not been adequately discussed. Therefore, in this study, we discussed in detail the relationship of the bioanalytical data from three different techniques, LC-MS/MS, ELISA, and microfluidic immunoassay, using 245 clinical plasma samples from non-small cell lung cancer patients treated with bevacizumab. The quantified concentration data of bevacizumab in human plasma indicated that the results obtained were almost the same correlation regardless of which technique was used. And the referable value from LC-MS/MS and microfluidic immunoassay were similar and correlated compared with ELISA.


bevacizumab bioanalysis ELISA LC-MS microfluidic immunoassay nSMOL 



This study of the LBA-based assay was performed by Dr. Naoe Yamane of CMIC Pharma Science (Tokyo, Japan).

Compliance with Ethical Standards

This study was reviewed and approved by the relevant institutional review boards and signed informed consent was obtained from all patients prior to participation. The procedures were in accordance with the Helsinki Declaration.

Supplementary material

12248_2019_369_MOESM1_ESM.docx (33 kb)
ESM 1 (DOCX 33 kb)


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Copyright information

© American Association of Pharmaceutical Scientists 2019

Authors and Affiliations

  1. 1.Leading Technology of Bioanalysis and Protein ChemistryShimadzu CorporationKyotoJapan
  2. 2.Shimadzu Bioscience Research PartnershipShimadzu Scientific InstrumentsBothellUSA
  3. 3.Research and Development Center for New Medical FrontiersKitasato University School of MedicineSagamiharaJapan
  4. 4.Division of Molecular PharmacologyNational Cancer Center Research InstituteTokyoJapan
  5. 5.Department of Pharmacology and Therapeutics, Fundamental Innovative Oncology CoreNational Cancer Center Research InstituteTokyoJapan
  6. 6.Division of Clinical Pharmacology and Translational Research, Exploratory Oncology Research & Clinical Trial CenterNational Cancer CenterTokyoJapan

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