Development of a general method for quantifying IgG-based therapeutic monoclonal antibodies in human plasma using protein G purification coupled with a two internal standard calibration strategy using LC-MS/MS

Huai Hsuan Chiu, Hsiao Wei Liao, Yu Yun Shao, Yen Shen Lu, Ching Hung Lin, I. Lin Tsai, Ching Hua Kuo

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Monoclonal antibody (mAb) drugs have generated much interest in recent years for treating various diseases. Immunoglobulin G (IgG) represents a high percentage of mAb drugs that have been approved by the Food and Drug Administration (FDA). To facilitate therapeutic drug monitoring and pharmacokinetic/pharmacodynamic studies, we developed a general liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to quantify the concentration of IgG-based mAbs in human plasma. Three IgG-based drugs (bevacizumab, nivolumab and pembrolizumab) were selected to demonstrate our method. Protein G beads were used for sample pretreatment due to their universal ability to trap IgG-based drugs. Surrogate peptides that were obtained after trypsin digestion were quantified by using LC-MS/MS. To calibrate sample preparation errors and matrix effects that occur during LC-MS/MS analysis, we used two internal standards (IS) method that include the IgG-based drug-IS tocilizumab and post-column infused IS. Using two internal standards was found to effectively improve quantification accuracy, which was within 15% for all mAb drugs that were tested at three different concentrations. This general method was validated in term of its precision, accuracy, linearity and sensitivity for 3 demonstration mAb drugs. The successful application of the method to clinical samples demonstrated its' applicability in clinical analysis. It is anticipated that this general method could be applied to other mAb-based drugs for use in precision medicine and clinical studies.

Original languageEnglish
JournalAnalytica Chimica Acta
DOIs
Publication statusAccepted/In press - Jan 1 2018

Fingerprint

Calibration
antibody
Purification
purification
Blood Proteins
drug
Immunoglobulin G
Monoclonal Antibodies
calibration
plasma
protein
Pharmaceutical Preparations
Therapeutics
Pharmacodynamics
Precision Medicine
Plasma (human)
method
Drug Monitoring
Pharmacokinetics
United States Food and Drug Administration

Keywords

  • Liquid chromatography-tandem mass spectrometry (LC-MS/MS)
  • Plasma
  • Post-column infusion internal standard (PCI-IS)
  • Protein G purification
  • Therapeutic monoclonal antibody (mAb)

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy

Cite this

Development of a general method for quantifying IgG-based therapeutic monoclonal antibodies in human plasma using protein G purification coupled with a two internal standard calibration strategy using LC-MS/MS. / Chiu, Huai Hsuan; Liao, Hsiao Wei; Shao, Yu Yun; Lu, Yen Shen; Lin, Ching Hung; Tsai, I. Lin; Kuo, Ching Hua.

In: Analytica Chimica Acta, 01.01.2018.

Research output: Contribution to journalArticle

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