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Overcoming Soluble Target Interference in Measurement of Total Bispecific Therapeutic Antibody Concentrations

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Abstract

The measurement of therapeutic drug concentrations is used to assess drug exposure and the relationship between therapeutic pharmacokinetics (PK) and pharmacodynamics (PD), which help determine the optimal dose for patients. Ligand binding assays (LBAs) are often the method of choice for evaluation of drug concentration and use either the therapeutic target protein or antibodies to the therapeutic as capture and/or detection reagents. Due to the bivalency of antibody therapeutics, heterogeneous states of the drug/target complex can exist in the presence of soluble targets which can complicate measurement of unbound drug. In the case of bispecific antibodies, measurement of drug can be even more complicated and depend upon the levels of both targets to each arm. Measuring the total drug allows for PKPD modeling prediction of human dose projections in addition to overcoming challenges associated with measuring free drug for bispecific antibodies. Here, we present a study in which a sandwich ELISA format was used to measure total anti-KLK5/KLK7 antibody concentrations. This assay utilized a non-blocking anti-idiotype (ID) antibody to one arm of the antibody for capture and an antibody to target bound to the other arm of the antibody for detection. Our qualified assay showed acceptable precision, accuracy, dilutional linearity, and reproducibility and enabled detection of a total bispecific antibody at high levels of two targets. To confirm that our assay was detecting total drug, a subset of samples was evaluated in a generic total LC–MS/MS assay.

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Acknowledgements

The authors would like to thank Wei Bu, Jason LaMar, and Rachel Basile for contributions to the mass spectrometry data and Paul Vu and Jose Diaz for providing the critical reagents for our experiments. Manuscript editing was provided by Anshin BioSolutions Corp.

Funding

All work described in this paper was funded by Genentech, Inc. The authors are all employees of Genentech, Inc., and stockholders of the F. Hoffmann-La Roche group.

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Authors and Affiliations

  1. BioAnalytical Sciences, Development Sciences, Genentech, Inc., 1 DNA Way, South San Francisco, California, 94080, USA

    Jeongsup Shim, Jessica Chen, Montserrat Carrasco-Triguero & Saloumeh K. Fischer

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Contributions

J.S.: conceptualization, data curation, formal analysis, methodology, project administration, and roles/writing—original draft; J.C.: data curation, formal analysis, methodology, and qualification; M.C-T.: project administration, writing—review and editing, and supervision; S.K.F.: conceptualization, project administration, writing—review and editing, and supervision.

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Correspondence to Jeongsup Shim.

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Shim, J., Chen, J., Carrasco-Triguero, M. et al. Overcoming Soluble Target Interference in Measurement of Total Bispecific Therapeutic Antibody Concentrations. AAPS J 25, 82 (2023). https://doi.org/10.1208/s12248-023-00848-9

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