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Automated Bioanalytical Workflow for Ligand Binding-Based Pharmacokinetic Assay Development.

Brad R Evans1, Armen G Beck2, Lai Yeung1

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|December 29, 2023
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Summary

Developing robust pharmacokinetic (PK) assays for therapeutic monoclonal antibodies (mAbs) is crucial. We created an automated platform to efficiently screen anti-idiotypic antibodies (anti-IDs) and assay formats, reducing time and resources for reliable drug concentration measurements.

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Area of Science:

  • Bioanalytical Chemistry
  • Pharmacokinetics
  • Monoclonal Antibody Therapeutics

Background:

  • Therapeutic monoclonal antibodies (mAbs) are increasingly important, necessitating accurate pharmacokinetic (PK) measurement in serum.
  • Ligand-binding assays using anti-idiotypic antibodies (anti-IDs) are common but can be affected by soluble antigens, leading to underestimation of drug levels.
  • Robust PK assays require anti-IDs with high affinity, minimal interference from soluble antigens, and low matrix effects.

Purpose of the Study:

  • To develop an integrated, automated screening and analysis platform for robust bioanalytical pharmacokinetic (PK) assay development.
  • To efficiently screen and select optimal anti-idiotypic antibodies (anti-IDs) and assay formats for therapeutic monoclonal antibody (mAb) quantification.
  • To reduce the time and resources required for developing reliable PK assays compared to traditional methods.

Main Methods:

  • Developed an automated workflow and scoring platform utilizing multiple bioanalytical assay parameters for ranking anti-idiotypic antibodies (anti-IDs).
  • Employed a primary indirect electrochemiluminescence (ECL) screen to shortlist anti-IDs for further testing.
  • Conducted a secondary ECL sandwich assay screen with labeled anti-ID pairings to evaluate multiple PK assay formats and identified the optimal anti-ID/assay combination.

Main Results:

  • Successfully developed an automated assay incorporating fixed plate maps and a human-guided graphical user interface (GUI) for scoring.
  • Compared the GUI-based scoring system with a data-dependent Gaussian mixture model scoring system for automated selection.
  • The automated approach significantly reduced the time and resources needed for screening anti-IDs and identifying the most robust PK assay format.

Conclusions:

  • The developed automated platform enables efficient screening of anti-idiotypic antibodies (anti-IDs) and identification of optimal pharmacokinetic (PK) assay formats.
  • This integrated approach enhances efficiency, quality, and robustness in bioanalytical assay development for therapeutic monoclonal antibodies (mAbs).
  • Standardized, automated platforms are critical for accelerating PK assay development to support future clinical trials.