Quantification of human mAbs in mouse tissues using generic affinity enrichment procedures and LC-MS detection

Bioanalysis. 2014;6(13):1795-811. doi: 10.4155/bio.14.143.

Abstract

Background: The disease state can modulate the penetration of large antibody-sized therapeutic molecules into affected tissues. Suitable bioanalytical methods are required for the quantitative analysis of drug tissue levels to enable a better understanding of the parameters influencing drug penetration and target engagement.

Results: Described is a sensitive and selective LC-MS/MS assay for the quantification of human mAb molecules in mouse tissues. By homogenizing tissues directly into serum, a common serum calibration curve can be used for multiple tissues. A generic procedure was used for affinity enrichment. An analytical range of 20 - 20,000 ng/ml was achieved in serum.

Conclusion: The method described here can be applied for the quantitative analysis of mAb and Fc-fusion therapeutic molecules in a variety of animal tissue matrices.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal / analysis*
  • Antibodies, Monoclonal / blood
  • Antibodies, Monoclonal / metabolism
  • Antibodies, Monoclonal, Humanized / analysis
  • Antibodies, Monoclonal, Humanized / blood
  • Antibodies, Monoclonal, Humanized / metabolism
  • Chromatography, Affinity
  • Chromatography, High Pressure Liquid*
  • Female
  • Humans
  • Liver / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Molecular Sequence Data
  • Peptides / analysis
  • Regression Analysis
  • Skin / metabolism
  • Tandem Mass Spectrometry*
  • Trypsin / metabolism
  • Ustekinumab

Substances

  • Antibodies, Monoclonal
  • Antibodies, Monoclonal, Humanized
  • Peptides
  • Trypsin
  • Ustekinumab