Development of a β-lactamase-based aggregation-induced emission lateral flow strip for the detection of clavulanic acid in Milk.

Lack of biorecognition elements significantly hinders the development of rapid detection methods for clavulanic acid (CA). To address this, we expressed Class A β-lactamases PC1 in vitro and demonstrated its high affinity for CA. Then we investigated the recognition mechanisms of PC1 for CA and identified key contact amino acids: Ser70, Lys73, Ser130, Glu166, and Lys234. Furthermore, PC1 was utilized as a novel biorecognition element to establish a "pseudo-immuno" lateral flow strip (LFS) for CA detection. Aggregation-induced emission fluorescence microspheres (AIE@FM) and biotin-streptavidin (Bio-SA) were integrated to improve the detection performance of PC1-based LFS. Results showed that the sensitivity (cut-off value) of PC1-based AIE(Bio-SA)-LFS was enhanced 2-fold and 4-fold compared to basic AIE@FM-LFS and traditional Au-based LFS, respectively. Eventually, the proposed PC1-based AIE(Bio-SA)-LFS was successfully verified in milk samples with a cut-off value of 20 ng mL. This study provides a powerful tool for on-site CA monitoring for the first time.