Evolution of high-affinity aptamer probe for label-free and probe conformation switch-based fluorescent biosensing of -carotene in seaweeds

beta-Carotene is an important bioactive component showing great activities and receiving great attention in recent years. The traditional methods for the accurate detection of beta-carotene in food are time-consuming. There is an urgent need to establish rapid quantification methods for beta-Carotene. Herein, a label-free fluorescent aptasensor which was based on the conformation-switch of the aptamer probe was developed to detect beta-carotene in seaweeds. Firstly, a post-SELEX strategy based on extraction and terminal-fixation was established to evolve the anti-beta-carotene aptamer to achieve high affinity. An aptamer probe (beta-2-TF) with a 177.5-fold affinity increment was obtained and then applied to develop a probe conformation-switch-based fluorescent aptasensor with the assistance of SYBR Green I (SGI). Under the optimal conditions, the aptasensor exhibited good accuracy and selectivity, with a linear detection range of 62.5-4000 nM and a limit of detection (LOD) as low as 39.64 nM. The developed strategies can act as robust references for the analytical method development in the food field.