Design of Molecularly Imprinted Polymer Nanoparticles Capable of Suppressing TEM-1 β-Lactamase Activity

Antimicrobial resistance (AMR) is a major global health threat, as classified by the World Health Organisation (WHO), and is driven by the ability of microorganisms to deactivate drugs. This research aims to address AMR by employing molecularly imprinted polymer nanoparticles (nano-MIPs) capable of inhibiting TEM-1 β-lactamase enzyme, a key contributor to bacterial drug resistance. We used a snapshot imprinting technique to map the epitopes of the TEM-1 enzyme. This mapping identified five crucial epitopes, which were then used to synthesize the nano-MIPs via a solid-phase protocol. The resulting nano-MIPs with 169-242 nm diameter, demonstrated exceptional affinity and selectivity for TEM-1, with dissociation constants (K _D ) as low as 0.006–0.35 nM. In vitro assays confirmed the effectiveness of the nano-MIPs in inhibiting 93% of the TEM-1 activity. We also investigated antibiotic resistance in both E. coli pET15b cell cultures and the culture supernatant, where TEM-1 is released. Initial tests revealed significant resistance to ampicillin in the culture supernatant, which nano-MIPs successfully mitigated. The nanoparticles were able to reduce the effective dose of the antibiotic from 31 µg×mL ^-1 to 15 µg×mL ^-1 . The high efficiency of nano-MIPs suggests that this approach could be broadly applied to target other critical biomarkers involved in bacterial survival, offering a promising new strategy to combat AMR.