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Quantifying mouse antibody drugs in mice is challenging. A new LC/MS/MS method using immunoaffinity enrichment and a unique peptide offers a sensitive and cost-effective solution for pharmacokinetic studies.

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Area of Science:

  • Immuno-oncology
  • Bioanalytical Chemistry
  • Pharmacokinetics

Background:

  • Surrogate mouse monoclonal antibodies are crucial for immuno-oncology research, requiring accurate quantification in mouse models.
  • Traditional ligand-binding assays (LBAs) face challenges with cross-reactivity and reagent availability for mouse antibody drug quantification.
  • High endogenous immunoglobulin levels in mice complicate bioanalytical method development.

Purpose of the Study:

  • To develop a sensitive and specific bioanalytical method for quantifying surrogate mouse antibody drugs in mouse plasma.
  • To overcome the limitations of LBAs when specific reagents are unavailable.
  • To establish a cost-effective method for pharmacokinetic (PK) studies.

Main Methods:

  • Development of an automated multicycle immunoaffinity enrichment method using a commercially available anti-mouse IgG1 secondary antibody.
  • Identification of a unique surrogate peptide from the mouse antibody sequence.
  • Quantification using liquid chromatography-tandem mass spectrometry (LC/MS/MS).
  • Reuse of the capture antibody up to six times to address binding capacity issues.

Main Results:

  • The LC/MS/MS assay achieved a limit of quantitation of 5 ng/mL.
  • The method demonstrated satisfactory assay precision, accuracy, and dynamic range.
  • The immunoaffinity enrichment step was made more cost-effective through antibody reuse.
  • The assay successfully quantified a surrogate mouse IgG1 (mIgG1) antibody drug in mouse plasma.

Conclusions:

  • A novel automated immunoaffinity enrichment LC/MS/MS assay enables sensitive quantification of mouse antibody drugs in mice.
  • This approach eliminates the need for specially generated immunoaffinity capturing reagents.
  • The method is suitable for discovery pharmacokinetic studies, improving mouse to human translation.