A novel biomarker assay qualification strategy for rare human matrices – a case study of biomarkers in aqueous humor

Abstract

Background: Measuring pharmacodynamic biomarkers near the therapeutic site of action presents considerable challenges for sites with limited matrix volume or difficult access. Bioanalytical method qualification requires the use of numerous matrix samples, which is problematic for rare matrices. The aim of this study was to design and implement a streamlined, fit-for-purpose strategy for qualification of biomarker assays in rare matrices. Materials & methods: A multiplexed biomarker immunoassay was developed in human aqueous humor. Results: Our strategy was successfully implemented, providing characterization of assay performance while reducing number of samples in assay qualification. Our assay was used in clinical trial support for an ophthalmic drug candidate. Conclusion: Our results indicate this approach can be applied to other early stage drug development programs facing similar challenges.

Tweetable abstract

Bioanalytical assay development and qualification requires the use of numerous matrix samples, which is problematic for rare matrices. We designed and successfully implemented a streamlined strategy for qualification of biomarker assays in rare matrices.

Summary points

• To address the challenge of a limited number and extremely low volume of rare matrix samples.

• We developed a streamline strategy to qualify clinical biomarkers assays in those matrices.

• This strategy was successfully applied to an Ella immunoassay measuring four cytokines in aqueous humor.

• The Ella immunoassay was subsequently used to support the pharmacodynamics analysis for a clinical trial of an ophthalmologic drug.

• All four cytokines were detectable at an equivalent frequency in a limited number of commercially sourced aqueous humor samples compared with an expanded panel.

• A similar streamlined biomarker assay qualification strategy can be applied to other rare human matrices.

Keywords: :

• aqueous humor
• biomarker assay qualification
• Ella
• pharmacodynamic biomarker
• rare matrix

Financial disclosure

Financial and material support from Genentech was received by all authors for this research and the creation of this work. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

Competing interests disclosure

All authors are employees of Genentech, Inc. and stockholders of the Roche group. The authors have no other competing interests or relevant affiliations with any organization or entity with the subject matter or materials discussed in the manuscript apart from those disclosed.

Writing disclosure

Medical writing support was provided by Anshin BioSolutions and was funded by Genentech.

Acknowledgments

The authors thank Vahan Indjeian, Michael Elliott, Wayne Kung, May Chen, Henry Wiley, Hao Chen and Surinder Kaur for valuable feedback on various topics addressed in this paper. They acknowledge Violet Lee for initial discussions on the bioanalytical strategy, and Jose Diaz for support in sourcing commercially-available aqueous humor samples.

Additional information

Funding

Financial and material support from Genentech was received by all authors for this research and the creation of this work.