Activated alkyne- enabled turn- on click bioconjugation with cascade signal amplification for ultrafast and high- throughput antibiotic screening

Antimicrobial susceptibility testing plays a pivotal role in the discovery of new antibi-otics. However, the development of simple, sensitive, and rapid assessment approaches remains challenging. Herein, we report an activated alkyne- based cascade signal ampli-fication strategy for ultrafast and high-throughput antibiotic screening. First of all, a novel water-soluble aggregation-induced emission (AIE) luminogen is synthesized, which contains an activated alkyne group to enable fluorescence turn -on and metal -free click bioconjugation under physiological conditions. Taking advantage of the in -house established method for bacterial lysis, a number of clickable biological substances (i.e., bacterial solutes and debris) are released from the bacterial bodies, which remarkably increases the quantity of analytes. By means of the activated alkyne-mediated turn -on click bioconjugation, the system fluorescence signal is significantly amplified due to the increased labeling sites as well as the AIE effect. Such a cascade signal amplification strategy efficiently improves the detection sensitivity and thus enables ultrafast antimi-crobial susceptibility assessment. By integration with a microplate reader, this approach is further applied to high-throughput antibiotic screening.

Automated summary

This article introduces an activated alkyne-based cascade signal amplification strategy for ultrafast and high-throughput antibiotic screening. By synthesizing a novel water-soluble aggregation-induced emission (AIE) luminogen with an activated alkyne group, fluorescence can be turned on and metal-free click bioconjugation can be achieved under physiological conditions. By utilizing the method of bacterial lysis, clickable biological substances are released, significantly increasing the quantity of analytes. The system fluorescence signal is greatly amplified through activated alkyne-mediated turn-on click bioconjugation. This cascade signal amplification strategy effectively improves the detection sensitivity and enables ultrafast antimicrobial susceptibility assessment. Integrated with a microplate reader, this approach can be applied to high-throughput antibiotic screening.