Development of an LC-MS/MS method with protein G purification strategy for quantifying bevacizumab in human plasma

Huai Hsuan Chiu, I. Lin Tsai, Yen Shen Lu, Ching Hung Lin, Ching Hua Kuo

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Biopharmaceutical products such as protein drugs and monoclonal antibodies (mAb) are currently of great interest with monoclonal antibody drugs being one of the fastest growing categories of biopharmaceutical products. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) has gained high interest for measuring mAb drugs in biological samples in recent years due to its high selectivity. Bevacizumab is a humanized immunoglobulin G (IgG) mAb drug against human vascular endothelial cell growth factor A (VEGF-A). It is used for treating many types of cancers. Recent studies have indicated that clinical outcomes vary among patients treated with bevacizumab and produce various side effects, such as vascular disorders. In this study, we developed an LC-MS/MS method to quantify bevacizumab concentration. We selected readily available and economic materials for sample preparation to facilitate its wider use in clinical fields.—Protein G was used to trap bevacizumab from human plasma. In place of an extended stable isotope-labeled internal standard (SIL-IS), the IgG-based drug-IS tocilizumab was used because of its better calibration performance. The method was validated in terms of its precision, accuracy, linearity, and sensitivity. The accuracies which were expressed as percentage recoveries for three concentration levels were within 92.8 ± 3.2 to 112.7 ± 4.5%. Repeatability and intermediate precision in terms of peak area ratios were lower than 5.2 and 12.9% RSD, respectively. The application to patients’ sample measurements revealed a wide individual variability of drug concentrations, and the proposed simple and general method may facilitate personalized medicine for improving therapeutic efficacy and safety. [Figure not available: see fulltext.]

Original languageEnglish
Pages (from-to)1-11
Number of pages11
JournalAnalytical and Bioanalytical Chemistry
DOIs
Publication statusAccepted/In press - Sep 29 2017

Fingerprint

Plasma (human)
Purification
Monoclonal Antibodies
Pharmaceutical Preparations
Proteins
Immunoglobulin G
Precision Medicine
Liquid chromatography
Endothelial cells
Cell growth
Tandem Mass Spectrometry
Liquid Chromatography
Isotopes
Calibration
Medicine
Mass spectrometry
Blood Vessels
Bevacizumab
Intercellular Signaling Peptides and Proteins
Endothelial Cells

Keywords

  • Bevacizumab
  • In-solution digestion
  • LC-MS/MS
  • Monoclonal antibody (mAb)
  • Plasma
  • Protein G purification

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

Cite this

Development of an LC-MS/MS method with protein G purification strategy for quantifying bevacizumab in human plasma. / Chiu, Huai Hsuan; Tsai, I. Lin; Lu, Yen Shen; Lin, Ching Hung; Kuo, Ching Hua.

In: Analytical and Bioanalytical Chemistry, 29.09.2017, p. 1-11.

Research output: Contribution to journalArticle

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