Conjugated Polymers Blended with Lipids and Galactosidase as Langmuir-Blodgett Films To Control the Biosensing Properties of Nanostructured Surfaces.

The structure of enzymes must be conserved when incorporated in nanoelectronic devices because their activity determines the function of the device as sensors. Among the systems that can retain their conformational structures, Langmuir-Blodgett (LB) films can be useful to exploit the construction of bioelectronic devices organized at the molecular level because biological and polymeric materials can be coupled as ultrathin films for biosensors and actuators. In this paper, we immobilized a ß-galactosidase enzyme in the LB films of stearic acid and the conjugated polymer poly[(9,9-dioctylfluorene)- co-thiophene]. After the characterization of the floating films using tensiometry, vibrational spectroscopy, and Brewster angle microscopy, they were transferred to solid supports as LB films, and the catalytic activity of the enzyme could be preserved as analyzed using UV-vis spectroscopy. We noted that the presence of a supramolecular structure formed in the LB films not only conserved the enzyme activity but also exhibited regular and distinctive output signals in all molecular architectures employed in this work. These results are related to the synergism between the compounds on the active layer associated with a surface morphology that facilitated the analyte diffusion because of an adequate molecular accommodation of all components. This work then demonstrates the viability of employing LB films composed of lipids, enzymes, and synthetic polymers as devices for biosensing applications.

Rhodanese incorporated in Langmuir and Langmuir-Blodgett films of dimyristoylphosphatidic acid: Physical chemical properties and improvement of the enzyme activity.

Preserving the catalytic activity of enzymes immobilized in bioelectronics devices is essential for optimal performance in biosensors. Therefore, ultrathin films in which the architecture can be controlled at the molecular level are of interest. In this work, the enzyme rhodanese was adsorbed onto Langmuir monolayers of the phospholipid dimyristoylphosphatidic acid and characterized by surface pressure-area isotherms, polarization-modulated infrared reflection-absorption spectroscopy (PM-IRRAS), and Brewster angle microscopy (BAM). The incorporation of the enzyme (5% in mol) in the lipid monolayer expanded the film, providing small surface domains, as visualized by BAM. Also, amide bands could be identified in the PM-IRRAS spectra, confirming the presence of the enzyme at the air-water interface. Structuring of the enzyme into α-helices was identified in the mixed monolayer and was preserved when the film was transferred from the liquid interface to solids supports as Langmuir-Blodgett (LB) films. The enzyme-lipid LB films were then characterized by fluorescence spectroscopy, PM-IRRAS, and atomic force microscopy. Measurements of the catalytic activity towards cyanide showed that the enzyme accommodated in the LB films preserved more than 87% of the enzyme activity in relation to the homogeneous medium. After 1 month, the enzyme in the LB film maintained 85% of the activity in contrast to the homogeneous medium, which 24% of the enzyme activity was kept. The method presented in this work not only points to an enhanced catalytic activity toward cyanide, but also may explain why certain film architectures exhibit an improved performance.