Metal-Organic-Framework-Based Enzymatic Microfluidic Biosensor via Surface Patterning and Biomineralization

Recently, the biomineralization of enzyme in metal-organic-framework (enzyme-MOF) composite have shown a great potential to increase enzymes stability without compromising their activity; hence, it is desirable for its applications in biosensing devices. Nonetheless, most of the enzyme-MOF research has been focusing on enzyme encapsulation in particle form, which limits its synthesis flexibility for practical applications because of its requirement for postsynthesis immobilization onto solid support. Therefore, to develop a diagnostic device out of the biomineralized enzyme, surface patterning and integration of microfluidic system offers many advantages. In this work, mussel-inspired polydopamine/polyethyleneimine (PDA/PEI) coating is employed to pattern enzyme-MOF in microfluidic channels and exploit the wettability gradient for pumpless transportation effect. As a proof of concept, we combine a cascade reaction of glucose oxidase (GOx) and horseradish peroxidase (HRP) enzymes to detect glucose into a patterned zeolitic imidazole framework-8 (ZIF-8) thin film on a flexible polymeric substrate. The results show that the ZIF-8/GOx&HRP in situ composites on PDA/PEI patterns have good acid and thermal stability compared with samples without ZIF-8. ZIF-8/GOx&HRP in situ shows high selectivity toward glucose, linear sensitivity of 0.00303 Abs/mu M, and the limit of detection of 8 mu M glucose concentration. An unexpected benefit of this approach is the ability of the ZIF-8 thin-film structure to provide a diffusion limiting effect for substrate influx, thus, producing high range of linear response range (8 mu M to 5 mM of glucose). This work provides insights into the spatial location of the enzymes in MOF thin films and the potential of such patterning techniques for MOF-based biosensors using other types of biological elements such as antibodies and aptamers.