Optimization of an ammonia assay based on transmembrane pH-gradient polymersomes.

ABSTRACT: Reliable ammonia quantification assays are essential for monitoring ammonemia in patients with liver diseases. In this study, we describe the development process of a microplate-based assay for accurate, precise, and robust ammonia quantification in biological fluids, following regulatory guidelines on bioanalytical method validation. The assay is based on transmembrane pH-gradient polymersomes that encapsulate a pH-sensitive ratiometric fluorophore, the fluorescence signal of which correlates with the ammonia concentration in the sample. Using a four-parameter logistic regression, the assay had a large quantification range (30-800 μM ammonia). As for selectivity, the presence of amino acids or pyruvate (up to clinically relevant concentrations) showed no assay interference. In samples wit

SUBMITTER: Spyrogianni A 

PROVIDER: S-EPMC8586157 | biostudies-literature | 2021 Nov

REPOSITORIES: biostudies-literature