Phage-targeting bimetallic nanoplasmonic biochip functionalized with bacterial outer membranes as a biorecognition element

The use of phages-a natural predator of bacteria-has emerged as a therapeutic strategy for treating multidrugresistant bacterial infections; thus, the isolation and detection of phages from the environment is crucial for advancing phage therapy. Herein, for the first time, we propose a nanoplasmonic-based biodetection platform for phages that utilizes bacterial outer membranes (OMs) as a biorecognition element. Conventional biosensors based on phage-bacteria interactions encounter multiple challenges due to the bacteriolytic phages and potentially toxic bacteria, resulting in instability and risk in the measurement. Therefore, instead of whole living bacteria, we employ a safe biochemical OMs fraction presenting phage-specific receptors, allowing the robust and reliable phage detection. In addition, the biochip is constructed on bimetallic nanoplasmonic islands through solid-state dewetting for synergy between Au and Ag, whereby sensitive detection of phage-OMs interactions is achieved by monitoring the absorption peak shift. For high detection performance, the nanoplasmonic chip is optimized by systematically investigating the morphological features, e.g., size and packing density of the nanoislands. Using our optimized device, phages are detected with high sensitivity (& GE;-104 plaques), specificity (little cross-reactivity), and affinity (stronger binding to the host OMs than anti-bacterial antibodies), further exhibiting the cell-killing activities.

Automated summary

The use of phages as a therapeutic strategy for multidrug-resistant bacterial infections requires the isolation and detection of phages from the environment. This study proposes a nanoplasmonic-based biodetection platform that utilizes bacterial outer membranes as a biorecognition element. By using a safe biochemical outer membrane fraction presenting phage-specific receptors, the robust and reliable detection of phages is achieved.