Pharmaceutical Research

, Volume 20, Issue 11, pp 1885–1900

Recommendations for the Bioanalytical Method Validation of Ligand-Binding Assays to Support Pharmacokinetic Assessments of Macromolecules

  • Binodh DeSilva
  • Wendell Smith
  • Russell Weiner
  • Marian Kelley
  • JoMarie Smolec
  • Ben Lee
  • Masood Khan
  • Richard Tacey
  • Howard Hill
  • Abbie Celniker


Purpose.With this publication a subcommittee of the AAPS Ligand Binding Assay Bioanalytical Focus Group (LBABFG) makes recommendations for the development, validation, and implementation of ligand binding assays (LBAs) that are intended to support pharmacokinetic and toxicokinetic assessments of macromolecules.

Methods. This subcommittee was comprised of 10 members representing Pharmaceutical, Biotechnology, and the contract research organization industries from the United States, Canada, and Europe. Each section of this consensus document addresses a specific analytical performance characteristic or aspect for validation of a LBA. Within each section the topics are organized by an assay's life cycle, the development phase, pre-study validation, and in-study validation. Because unique issues often accompany bioanalytical assays for macromolecules, this document should be viewed as a guide for designing and conducting the validation of ligand binding assays.

Results. Values of ±20% (25% at the lower limit of quantification [LLOQ]) are recommended as default acceptance criteria for accuracy (% relative error [RE], mean bias) and interbatch precision (%coefficient of variation [CV]). In addition, we propose as secondary criteria for method acceptance that the sum of the interbatch precision (%CV) and the absolute value of the mean bias (%RE) be less than or equal to 30%. This added criterion is recommended to help ensure that in-study runs of test samples will meet the proposed run acceptance criteria of 4-6-30. Exceptions to the proposed process and acceptance criteria are appropriate when accompanied by a sound scientific rationale.

Conclusions. In this consensus document, we attempt to make recommendations that are based on bioanalytical best practices and statistical thinking for development and validation of LBAs.

best practices bioanalytical assay biological matrices consensus document immunoassay 


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Copyright information

© Plenum Publishing Corporation 2003

Authors and Affiliations

  • Binodh DeSilva
    • 1
  • Wendell Smith
    • 2
  • Russell Weiner
    • 3
  • Marian Kelley
    • 4
  • JoMarie Smolec
    • 5
  • Ben Lee
    • 6
  • Masood Khan
    • 7
  • Richard Tacey
    • 8
  • Howard Hill
    • 9
  • Abbie Celniker
    • 10
  1. 1.Amgen IncThousand Oaks
  2. 2.Lilly Research LaboratoriesGreenfield
  3. 3.Bristol-Myers SquibbPrinceton
  4. 4.Johnson and Johnson PharmaceuticalRaritan
  5. 5.Alta Analytical LaboratorySan Diego
  6. 6.Pfizer Global Research and DevelopmentAnn Arbor
  7. 7.Covance LaboratoriesChantilly
  8. 8.PPD DevelopmentRichmondVirginia
  9. 9.HLSCambridgeshireUK
  10. 10.Millennium PharmaceuticalsCambridge

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