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

Analytical and Bioanalytical Chemistry volume 409, pages 6583–6593 (2017)Cite this article

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.

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Acknowledgments

This study was supported by the Ministry of Science and Technology, Taiwan (MOST 105-2113-M-002-013-). The authors thank the NTU Integrated Core Facility for Functional Genomics of the National Research Program for Genomic Medicine of Taiwan for the technical assistance.

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Authors and Affiliations

  1. School of Pharmacy, College of Medicine, National Taiwan University, No. 33, Linsen S. Rd., Zhongzheng District, Taipei, 100, Taiwan, Republic of China

    Huai-Hsuan Chiu & Ching-Hua Kuo

  2. Department of Biochemistry and Molecular Cell Biology, School of Medicine, College of Medicine, Taipei Medical University, No. 250, Wu-Hsing Street, Taipei, 110, Taiwan, Republic of China

    I-Lin Tsai

  3. Department of Oncology, National Taiwan University Hospital, No. 7 Zhongshan S. Rd., Zhongzheng District, Taipei, 100, Taiwan, Republic of China

    Yen-Shen Lu & Ching-Hung Lin

  4. Department of Internal Medicine, National Taiwan University Hospital, Zhongzheng District, Taipei, 100, Taiwan, Republic of China

    Ching-Hung Lin

  5. The Metabolomics Core Laboratory, Center of Genomic Medicine, National Taiwan University, No. 2 Syu-jhou Road, Taipei, 10055, Taiwan, Republic of China

    Ching-Hua Kuo

  6. Department of Pharmacy, National Taiwan University Hospital, No. 33, Linsen S. Rd., Zhongzheng District, Taipei, 100, Taiwan, Republic of China

    Ching-Hua Kuo

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  1. Huai-Hsuan Chiu
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  2. I-Lin Tsai
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Correspondence to Ching-Hua Kuo.

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Plasma samples including healthy volunteers and patients were collected at the National Taiwan University Hospital and obtained with informed consent. The study was approved by the institutional review board of the National Taiwan University Hospital.

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The authors declare that they have no conflict of interest.

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Chiu, HH., Tsai, IL., Lu, YS. et al. Development of an LC-MS/MS method with protein G purification strategy for quantifying bevacizumab in human plasma. Anal Bioanal Chem 409, 6583–6593 (2017). https://doi.org/10.1007/s00216-017-0607-0

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  • DOI: https://doi.org/10.1007/s00216-017-0607-0

Keywords

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