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Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms
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Bioanalytical platform comparison using a generic human IgG PK assay format.

Beth A Leary1, Rosemary Lawrence-Henderson, Carolyn Mallozzi

  • 1Department of Pharmacokinetics, Dynamics and Metabolism Pfizer Inc, One Burtt Road, Andover, MA 01810, USA.

Journal of Immunological Methods
|September 3, 2013
PubMed
Summary
This summary is machine-generated.

This study compared four ligand-binding assay platforms for quantifying human IgG₁ monoclonal antibodies (MAbs) in rat serum. Meso Scale Discovery and Gyros platforms demonstrated superior performance, becoming the preferred choice for discovery support.

Keywords:
Generic IgG assayImmunoaffinityImmunoassayLigand-binding assay (LBA)Mass spectrometry

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Immunology

Background:

  • Accurate quantification of monoclonal antibodies (MAbs) is crucial for biotherapeutic development.
  • Ligand-binding assay (LBA) platforms offer various technologies for MAb measurement.
  • Selecting an optimal platform impacts assay performance and resource allocation.

Purpose of the Study:

  • To compare the performance of four LBA platforms (ELISA, Meso Scale Discovery, Gyros, AlphaLISA) for human IgG₁ MAb quantification in rat serum.
  • To identify the most suitable platform for supporting non-regulated (discovery) studies.
  • To evaluate immunoaffinity LC-MS/MS as an alternative quantitation method.

Main Methods:

  • Quantitative assays were developed and optimized for each platform using Fc-specific reagents.
  • Human IgG₁ MAb levels were measured in rat serum using mock samples across all platforms.
  • Assay parameters including sensitivity, dynamic range, MRL, sample volume, cost, and equipment were evaluated.
  • Immunoaffinity LC-MS/MS was explored for generic Fc quantitation.

Main Results:

  • Meso Scale Discovery and Gyros platforms exhibited superior assay performance compared to ELISA and AlphaLISA.
  • These two platforms were selected as the default choice for discovery study support.
  • Immunoaffinity LC-MS/MS demonstrated comparable performance to LBAs for generic Fc quantitation.

Conclusions:

  • Meso Scale Discovery and Gyros are recommended as default platforms for quantifying monoclonal antibodies in discovery settings.
  • Platform selection should be based on assay performance, cost, and available resources.
  • Immunoaffinity LC-MS/MS serves as a viable alternative for Fc quantitation.