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Fit-for-Purpose Validation of a Ligand Binding Assay for Toxicokinetic Study Using Mouse Serial Sampling

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Abstract

Purpose

The purpose of this study was to validate a ligand binding assay for the quantitation of a monoclonal antibody-based biotherapeutics (PF-57781346) in samples collected via capillary microsampling to support a regulated mouse toxicity study.

Method

A quantitative ligand binding assay on the Gyrolab platform was developed to quantify PF-57781346 in blood samples derived from capillary mouse serial sampling. The method validation evaluated assay characteristics including accuracy and precision, influence of sample processing on drug quantitation, whole blood matrix selectivity, dilution linearity and the stability of the drug in the study sample matrix.

Results

The method validation demonstrated acceptable analytical characteristics. The whole blood selectivity testing demonstrated accuracy between −4.8% and 13.9% in 10 out of 10 individual whole blood samples, suggesting that drug quantitation from whole blood is not impacted by the serial sampling procedure. Short-term and long-term drug stability in study sample matrix were established to cover required stability for sample storage and analysis (accuracy between −7.3% and 6.1%).

Conclusion

We reported a successful validation of a bioanalytical method that quantifies PF-55781346 in samples collected via capillary microsampling. The experience shared in this study could serve as a model process for bioanalytical method validation when capillary microsampling is used.

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Abbreviations

CMS:

Capillary microsampling

CV:

Coefficient of variation

DBS:

Dried blood spot

DQC:

Dilution quality control

ELISA:

Enzyme-linked immunosorbent assay

GLP:

Good laboratory practices

HQC:

High quality control

LBA:

Ligand binding assay

LC-MS:

Liquid chromatography-mass spectrometry

LIMS:

Laboratory information management system

LLOQ:

Lower limit of quantitation

LQC:

Low quality control

LTS:

Long-term stability

MQC:

Middle level quality control

MRD:

Minimum required dilution

PK:

Pharmacokinetics

PMT:

Photomultiplier Tube

QC:

Quality control

RE:

Relative error

ROQ:

Range of quantitation

RT:

Room temperature

TK:

Toxicokinetics

ULOQ:

Upper limit of quantitation

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Acknowledgments and Disclosures

The authors would like to thank Soma Basak and Brianna Donnelly for performing validation experiments, Teresa Caiazzo, Renee Ramsey, Lee Walrus, and Glenn Miller for reagent support, and Yan Weng, Marcela Araya Roldan, Nicole Duriga, Katrina Olsen, and Courtnie Vennix for helpful discussions on the design of the current validation.

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Authors and Affiliations

  1. Biomedicine Design, Worldwide Research & Development, Pfizer Inc., One Burtt Raod, Andover, MA, 01810, USA

    Liang Zhu, Ying Wang, Alison Joyce, Ihor Djura & Boris Gorovits

Authors
  1. Liang Zhu
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  2. Ying Wang
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  3. Alison Joyce
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  4. Ihor Djura
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  5. Boris Gorovits
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Correspondence to Liang Zhu.

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Zhu, L., Wang, Y., Joyce, A. et al. Fit-for-Purpose Validation of a Ligand Binding Assay for Toxicokinetic Study Using Mouse Serial Sampling. Pharm Res 36, 169 (2019). https://doi.org/10.1007/s11095-019-2699-z

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  • DOI: https://doi.org/10.1007/s11095-019-2699-z

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