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Bioanalytical Approaches to Quantify “Total” and “Free” Therapeutic Antibodies and Their Targets: Technical Challenges and PK/PD Applications Over the Course of Drug Development

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Abstract

The predominant driver of bioanalysis in supporting drug development is the intended use of the data. Ligand-binding assays (LBA) are widely used for the analysis of protein biotherapeutics and target ligands (L) to support pharmacokinetics/pharmacodynamics (PK/PD) and safety assessments. For monoclonal antibody drugs (mAb), in particular, which non-covalently bind to L, multiple forms of mAb and L can exist in vivo, including free mAb, free L, and mono- and/or bivalent complexes of mAb and L. Given the complexity of the dynamic binding equilibrium occurring in the body after dosing and multiple sources of perturbation of the equilibrium during bioanalysis, it is clear that ex vivo quantification of the forms of interest (free, bound, or total mAb and L) may differ from the actual ones in vivo. LBA reagents and assay formats can be designed in principle to measure the total or free forms of mAb and L. However, confirmation of the forms being measured under the specified conditions can be technically challenging. The assay forms and issues must be clearly communicated and understood appropriately by all stakeholders as the program proceeds through the development process. This paper focuses on monoclonal antibody biotherapeutics and their circulatory L that are either secreted as soluble forms or shed from membrane receptors. It presents an investigation into the theoretical and practical considerations for total/free analyte assessment to increase awareness in the scientific community and offer bioanalytical approaches to provide appropriate PK/PD information required at specific phases of drug development.

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Abbreviations

ADA:

Anti-drug antibodies

Anti-id:

Anti-idiotypic

CDR:

Complementarity-determining regions

ELISA:

Enzyme-linked immunosorbent assay

FIH:

First in human

Fc:

Fragment cystallizable region

IC50 :

The concentration of the interfering compound at which the analyte recovery is 50% of that in the absence of the interfering compound (50% inhibition point)

IgG:

Immunoglobulin G

K d :

Dissociation equilibrium constant

LBA:

Ligand-binding assay

L:

Target ligand

Lfree :

Target ligand not bound to drug

Ltotal :

Sum of bound and unbound target ligand

mAb:

Monoclonal antibody

mAbfree :

The unbound and partially free forms of the monoclonal antibody drug that are able to bind to the target ligand to mediate biological actions

mAbtotal :

Sum of bound partially bound and unbound forms of the monoclonal antibody drug

mAb*L:

Complex of monoclonal antibody drug and target ligand

pAb:

Polyclonal antibodies

PD:

Pharmacodynamics

PK:

Pharmacokinetics

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Authors and Affiliations

  1. Amgen Inc, One Amgen Center Drive, Thousand Oaks, California, 91320, USA

    Jean W. Lee, Hossein Salimi-Moosavi & Jian-Feng Lu

  2. MKelley Consulting LLC, West Chester, Pennsylvania, USA

    Marian Kelley

  3. Pfizer Inc, Groton, Connecticut, USA

    Lindsay E. King

  4. Genentech, South San Francisco, California, USA

    Jihong Yang & Meina T. Tang

  5. Covance, Princeton, New Jersey, USA

    John Kamerud

  6. Pharmargo LLC, Union City, California, USA

    Ago Ahene

  7. Bristol-Myers Squibb, Princeton, New Jersey, USA

    Heather Myler

  8. Genzyme, Framingham, Massachusetts, USA

    Cindy Rogers

Authors
  1. Jean W. Lee
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  2. Marian Kelley
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  3. Lindsay E. King
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  4. Jihong Yang
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  7. Jian-Feng Lu
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  9. Ago Ahene
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  10. Heather Myler
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  11. Cindy Rogers
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Correspondence to Jean W. Lee.

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Lee, J.W., Kelley, M., King, L.E. et al. Bioanalytical Approaches to Quantify “Total” and “Free” Therapeutic Antibodies and Their Targets: Technical Challenges and PK/PD Applications Over the Course of Drug Development. AAPS J 13, 99–110 (2011). https://doi.org/10.1208/s12248-011-9251-3

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  • DOI: https://doi.org/10.1208/s12248-011-9251-3

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