What sample preparation should be chosen for targeted MS monoclonal antibody quantification in human serum?

Bioanalysis. 2018 May 1;10(10):723-735. doi: 10.4155/bio-2018-0027. Epub 2018 May 17.

Abstract

Aim: Monoclonal antibody-based treatment of cancer has been established as one of the most successful therapeutic strategies.

Materials & methods: In this work, we developed a workflow based on an automated protein-A capture and LC-MS/MS analysis to quantify bevacizumab on patient serum during treatment. This analytical approach was fully validated and compared with a commercially available Monoclonal antibody-based treatment preparation (nanosurface and molecular-orientation limited kit).

Results: The analytical comparison of the two preparative workflows based on protein-A capture gave similar results with a better lower limit of quantification for the nanosurface and molecular-orientation limited kit (0.26986 vs 1.9565 μg/ml).

Conclusion: LC-MS/MS has clear advantages compared with ELISA when considering method development time, multiplexing capacities and absolute quantification with internal standardization.

Keywords: MS; automation; bevacizumab; nSMOL; protein-A purification; therapeutic drug monitoring.

MeSH terms

  • Amino Acid Sequence
  • Analytic Sample Preparation Methods / methods*
  • Antibodies, Monoclonal / blood*
  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal / isolation & purification
  • Chromatography, Liquid
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Limit of Detection
  • Mass Spectrometry*
  • Proteolysis
  • Staphylococcal Protein A / immunology

Substances

  • Antibodies, Monoclonal
  • Staphylococcal Protein A