A multiplexed hybrid LC-MS/MS pharmacokinetic assay to measure two co-administered monoclonal antibodies in a clinical study

Bioanalysis. 2014;6(13):1781-94. doi: 10.4155/bio.14.142.

Abstract

Background: Combination therapies with monoclonal antibodies (mAbs) enhance therapeutic activity and may circumvent drug resistance. However, these studies present bioanalytical challenges for ligand-binding assays (LBAs). Recent MS-based protein quantification offers an alternative for bioanalysis.

Results: A hybrid LC-MS/MS assay was developed to simultaneously measure human serum concentrations of two mAbs. Anti-idiotypic reagents that did not work in LBAs were successfully used for mAb affinity capture enrichment. Stable isotope-labeled peptide internal standards were employed. The mAb quantification involved measuring a signature CDR peptide derived from each mAb as a surrogate. Selected clinical samples were successfully analyzed.

Conclusion: The multiplexed LC-MS/MS method provided a powerful quantitative tool for clinical PK assessment of co-administered mAbs without the requirement for stringent affinity capture reagents.

MeSH terms

  • Amino Acid Sequence
  • Antibodies, Monoclonal / blood*
  • Antibodies, Monoclonal / metabolism
  • Antibodies, Monoclonal / pharmacokinetics
  • Chromatography, High Pressure Liquid / standards
  • Humans
  • Immunoglobulin Light Chains / chemistry
  • Immunoglobulin Light Chains / metabolism
  • Isotope Labeling
  • Molecular Sequence Data
  • Peptides / analysis
  • Peptides / chemistry
  • Peptides / standards
  • Quality Control
  • Recombinant Proteins / blood
  • Recombinant Proteins / metabolism
  • Recombinant Proteins / pharmacokinetics
  • Tandem Mass Spectrometry* / standards

Substances

  • Antibodies, Monoclonal
  • Immunoglobulin Light Chains
  • Peptides
  • Recombinant Proteins