Optimizing Bacteriophage Surface Densities for Bacterial Capture and Sensing in QCM-D.

Surface immobilized bacteriophages (phages) are increasingly used as biorecognition elements on bacterial biosensors (e.g., on acoustical, electrical, or optical platforms). The phage surface density is a critical factor determining a sensor's bacterial binding efficiencies; in fact, phage surface densities that are too low or too high can result in significantly reduced bacterial binding capacities. Identifying an optimum phage surface density is thus crucial when exploiting the bacteriophages' bacterial capture capabilities in biosensing applications. Herein, we investigated surface immobilization of the Pseudomonas aeruginosa specific E79 (tailed) phage and the Salmonella Typhimurium specific PRD1 (non-tailed) phage and their subsequent bacterial capture abilities using quartz crystal microbalance with dissipation monitoring (QCM-D). The QCM-D was used in two experimental set-ups, (i) a conventional set-up, and (ii) a combined setup with ellipsometry. Both setups were exploited to link the phages' immobilization behaviors to their bacterium capture efficiency. While E79 displayed characteristic optima in both the mechanical (QCM-D) and the optical (ellipsometry) data that coincided with its specific bacterial capture optimum, no optima were observed during PRD1 immobilization. The characteristic optima suggests that the E79 phage undergoes a surface rearrangement event that changes the hydration state of the phage film, thereby impairing the E79 phage's ability to capture bacteria. However, the absence of such optima during deposition of the non-tailed PRD1 phage suggests that other mechanisms may also lead to reduced bacterial capture by surface immobilized bacteriophages.