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Development of a Highly Specific Anti-drug Antibody Assay in Support of a Nanoparticle-based Therapeutic

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Abstract

PEGylated biotherapeutics can elicit anti-PEG (polyethylene glycol) immune responses in patients treated with this category of drugs. While anti-PEG antibody assays for this class of biotherapeutics have become a common element of the clinical immunogenicity testing strategy, the overall antibody incidence induced by the nanoparticle (NP) delivery system (such as ACCURINS®) has not been fully studied to date. To support the immunogenicity assessment of one of Pfizer’s NP-based therapeutics, consisting of gedatolisib (GEDA) encapsulated in ACCURINS® (GEDA-NP), we developed an anti-GEDA-NP antibody (ADA) assay on the MSD platform for the detection of GEDA-NP induced ADA in human serum. The focus of our strategy was on developing a clinically relevant ADA assay and systematically addressing assay interference through rigorous assay optimization. Our efforts led to a fit-for-purpose assay for the detection of anti-GEDA-NP ADA in serum samples obtained from breast cancer patients. Results from method qualification indicated robust assay performance, as highlighted by inter and intra-assay precision within 25% CV for all controls, and reproducible response profiles across multiple runs during the assessment of assay cut points with breast cancer samples. The assay sensitivity was between 4.3 ng/mL and 123 ng/mL for surrogate positive controls of IgG and IgM isotypes, respectively. Additionally, assay interference from nonspecific matrix proteins and circulating drug was addressed, which ensured accurate assessment of ADA incidence that can be attributed to GEDA-NP.

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Abbreviations

PLGA:

Poly(lactide-co-glycolide)

PLA:

Poly(l-lactide)

PEG:

Polyethylene glycol

BGG:

Bovine gamma globulin

MRD:

Minimal required dilutions

ADA:

Anti-drug antibody

GEDA-NP:

ACCURINS encapsulated with gedatolisib (GEDA), gedatolisib is a dual inhibitor of PI3K and mTOR activity. Gedatolisib has been evaluated for the treatment of ER+/Her2− breast cancer in clinical studies

API:

Active pharmaceutical ingredient

PI3K:

Phosphatidylinositol-3-kinase (PI3K)

mTOR:

Mammalian target of rapamycin (mTOR)

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Acknowledgments

We thank Pfizer colleagues supported this work: Dr. Ravi Visswanathan for GEDA-NP program support; Shuenn Shyong Liou and Brianna Donnelly who executed this work with Judith F. Smith together; Meghana Deshpande for clinical assay implement logistics, and Pfizer’s Critical Reagent Group for coordination of reagent productions. In addition, we also appreciate Dr. Steven Max for his critical guidance during the process of this manuscript and Dr. John Kamerud and Mr. Michael Luong for critical review. Lastly, we want to thank our CRO colleagues who performed feasibility study of this method.

Funding

This work was exclusively supported by Pfizer Inc.

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Authors and Affiliations

  1. BioMedicine Design, Pfizer Inc, One Burtt Road, Andover, Massachusetts, 01810, USA

    Ying Wang, Judith F. Smith, Marcela M. Araya & Boris Gorovits

  2. Early Clinical Development, Pfizer Inc, 10777 Science Center Drive, San Diego, California, 92121, USA

    Kai-Hsin (Ken) Liao

Authors
  1. Ying Wang
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  2. Judith F. Smith
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  3. Marcela M. Araya
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  4. Kai-Hsin (Ken) Liao
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  5. Boris Gorovits
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Corresponding author

Correspondence to Ying Wang.

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Conflict of Interest

This study was funded by Pfizer Inc. All the authors listed are employees of the Pfizer Inc. The publication of this work has been cleared by Pfizer’s Manuscript Review for Publications and the content of this manuscript has not been published elsewhere.

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Wang, Y., Smith, J.F., Araya, M.M. et al. Development of a Highly Specific Anti-drug Antibody Assay in Support of a Nanoparticle-based Therapeutic. AAPS J 22, 81 (2020). https://doi.org/10.1208/s12248-020-00462-z

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  • DOI: https://doi.org/10.1208/s12248-020-00462-z

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