Enhancing the performance of an in vitro RNA biosensor through iterative design of experiments.

ABSTRACT: The quality control of RNA has become increasingly crucial with the rise of mRNA-based vaccines and therapeutics. However, conventional methods such as LC-MS often require specialized equipment and expertise, limiting their applicability to high throughput experiments. Here, we optimize a previously characterized RNA integrity biosensor, that provides a simple colorimetric output, using Design of Experiments (DoE). Through iterative rounds of a Definitive Screening Design (DSD) and experimental validation, we systematically explored different assay conditions to enhance the biosensor's performance. Optimization led to a 4.1-fold increase in dynamic range and reduced RNA concentration requirements by one-third, significantly improving usability. Notable modifications included reducing the c

SUBMITTER: Aw R 

PROVIDER: S-EPMC12348311 | biostudies-literature | 2025 Jul-Aug

REPOSITORIES: biostudies-literature