Three-Component Langmuir–Blodgett Films Consisting of Surfactant, Clay Mineral, and Lysozyme: Construction and Characterization



The Langmuir–Blodgett (L–B) technique has been employed for the construction of hybrid films consisting of three components: surfactant, clay, and lysozyme (Lys). The surfactants are octadecylammonium chloride (ODAH) and octadecyl ester of rhodamine B (RhB18). The clays include saponite and laponite. Surface pressure versus area isotherms indicate that lysozyme is adsorbed by the surfactant–clay L–B film at the air–water interface without phase transition. The UV-visible spectra of the hybrid film ODAH–saponite–Lys show that the amount of immobilized lysozyme in the hybrid film is (1.3±0.2) ng mm−2. The average surface area (Ω) per molecule of lysozyme is approximately 18.2 nm2 in the saponite layer. For the multilayer film (ODAH–saponite–Lys)n, the average amount of lysozyme per layer is (1.0±0.1) ng mm−2. The amount of lysozyme found in the hybrid films of ODAH–laponite–Lys is at the detection limit of about 0.4 ng mm−2. Attenuated total reflectance (ATR) FTIR spectra give evidence for clay layers, ODAH, lysozyme, and water in the hybrid film. The octadecylammonium cations are partially oxidized to the corresponding carbamate. A weak 1620 cm−1 band of lysozyme in the hybrid films is reminiscent of the presence of lysozyme aggregates. AFM reveals evidence of randomly oriented saponite layers of various sizes and shapes. Individual lysozyme molecules are not resolved, but aggregates of about 20 nm in diameter are clearly seen. Some aggregates are in contact with the clay mineral layers, others are not. These aggregates are aligned in films deposited at a surface pressure of 20 mN m−1.