Characterization and optimization of bioluminescent bacterial cells immobilization process in calcium alginate hydrogel tablets.

Abstract

Whole-cell biosensors (WCB) are increasingly utilized in various applications. These platforms integrate cells with a signal measurement device. One of the main challenges in the development of such platforms is the immobilization matrix that is used to keep the cells stable, which also affects the portability of the device. In this study, a portable and simple immobilization of bioluminescent bacterial cells in calcium alginate hydrogel was examined. The effects of several physical parameters were investigated (e.g. calcium alginate solution volume, drying, incubation time, mixing procedure, bacterial concentration, and tablet location within the cylinder). An alginate solution volume of 3mL was preferred as well as the addition of 400µL solution after the 15 minute of compressing step and before the polymerization step. Also, a stirring mixing mode is favored over vortexing due to the creation of better homogenized tablets, as well as a bacterial concentration of 0.15 OD600nm that produced a high light response while maintaining lower variance. Lastly, the findings showed a significantly higher response (Induction Factor (IF)) in the tablets using the optimized immobilization protocol (IF=8.814) than the old one (IF=1.979). To conclude, bacterial cells immobilization in calcium alginate tablets provides improved sensitivity and storability.