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Direct Measurement of KDM1A Target Engagement Using Chemoprobe-based Immunoassays
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Direct Measurement of KDM1A Target Engagement Using Chemoprobe-based Immunoassays

Published on: June 13, 2019

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Immunocapture-LC/MS-Based Target Engagement Measurement in Tumor Plasma Membrane

Hiroshi Sugimoto, Dhimankrishna Ghosh, Susan Chen

    Analytical Chemistry
    |October 30, 2018
    PubMed

    Insights

    A new assay platform using immunocapture-liquid chromatography mass spectrometry (IC-LC/MS) measures drug target engagement directly on tumor cell membranes. This method enhances understanding of drug efficacy for targeted therapies.

    Area of Science:

    • Biomedical Science
    • Analytical Chemistry
    • Pharmacology

    Background:

    • Immunocapture-liquid chromatography mass spectrometry (IC-LC/MS) is crucial for assessing drug exposure and target engagement in targeted therapies.
    • Current methods often analyze systemic circulation or tissue homogenates, but direct measurement at the cell plasma membrane, the primary drug action site, is preferred.
    • Existing IC-LC/MS methodologies for plasma membrane analysis in tissue samples are limited.

    Purpose of the Study:

    • To develop and validate an IC-LC/MS assay platform for quantifying target engagement specifically at the tumor plasma membrane.
    • To assess the in vivo target engagement of an antibody-drug conjugate (ADC) targeting guanylyl cyclase C (GCC) in tumor samples.
    • To improve the understanding of pharmacokinetic/pharmacodynamic (PK/PD) relationships for enhanced antitumor efficacy.

    Main Methods:

    • Development of an IC-LC/MS assay platform for analyzing tumor plasma membrane preparations.
    • Utilizing fully human IgG1 monoclonal antibody-based ADC treatment in tumor samples expressing GCC.
    • Differentiating between total and target-drug bound GCC fractions to calculate in vivo target engagement with minimal equilibrium shift.

    Main Results:

    • Successfully established an IC-LC/MS assay platform for tumor plasma membrane analysis.
    • Demonstrated the ability to accurately measure in vivo target engagement of an ADC targeting GCC.
    • The method minimizes potential equilibrium shifts between cell surface and organelle proteins.

    Conclusions:

    • The developed IC-LC/MS assay provides a robust method for assessing in vivo target engagement at the tumor plasma membrane.
    • This approach offers a more precise understanding of PK/PD relationships for targeted therapies.
    • The methodology has the potential to optimize therapeutic strategies for improved antitumor efficacy.

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