Kinetics Study of Bungarus fasciatus Venom Acetylcholinesterase Immobilised on a Langmuir–Blodgett Proteo‐Glycolipidic Bilayer

Abstract

This study deals with the kinetics properties of an enzyme immobilised in a defined orientation in a biomimetic environment. For this purpose, acetylcholinesterase (AChE) was captured at the surface of a nanostructured proteo-glycolipidic Langmuir-Blodgett film through specific recognition by a noninhibitor monoclonal antibody (IgG) inserted in a neoglycolipid bilayer. Modelling of this molecular assembly provided a plausible interpretation of the functional orientation of the enzyme. The AChE activity being stable for several weeks, the enzyme kinetics were investigated, and fitted perfectly with heterogeneous biocatalytic behaviour representative of cellular enzymatic catalysis. The AChE-IgG-glycolipid nanostructure was directly interfaced with an efficient optical device. Such an association, leading to an intimate contact between the nanostructure and the biochemical signal transducer, gives direct access to the intrinsic AChE behaviour. This study thus demonstrates the potential for direct investigation of the kinetic behaviour of an immobilised enzyme on a lipid bilayer through an efficient transduction system.

Reconstitution of Fusion Proteins in Supported Lipid Bilayers for the Study of Cell Surface Receptor–Ligand Interactions in Cell–Cell Contact

“…The simplest picture one can draw is that the Fc-part is embedded into the SLB while EA5-domains are situated outside (Figure 7A). 26 Because the Fc-domain requires more space (see ref 27) than the thickness of the SLB, it would be also partially exposed to the medium. Our antibody binding experiments show that both EA5-and the Fc-parts are accessible to the antibodies.…”

Self-assembly of DNA networks at the air–water interface over time

“…Even today, the LB technique is still an unique and important technique because it is the simplest and the most reliable method for obtaining ultra-thin films, with the highest material transferring efficiency. [23][24][25][26][27] The LB technique is also widely accepted as the most controllable method to transfer a surfactant monolayer from an air-water interface to a solid surface with least loss of the original molecular arrangements. 28,29 Therefore, it is broadly applied for obtaining physiologically compatible templates, 30,31 fabricating nanoscale-ordered chemical sensors, 28 and modifying electrodes or molecular electronic devices 27 with permeation selectivity.…”

Immunosensor for Pancreatic Cancer Based on Electrospun Nanofibers Coated with Carbon Nanotubes or Gold Nanoparticles

“…This requirement is normally fulfilled by controlling the molecular architecture of the nanostructured films of which most immunosensors are made. − In addition to the active layer containing antibodies or antigens, the nanostructured films contain a matrix designed to assist in preserving bioactivity and other components (e.g., nanomaterials) to enhance sensitivity . Experimental methods such as the self-assembled monolayer, , layer-by-layer, and Langmuir–Blodgett − techniques have been employed owing to their suitability to build complex molecular architectures, where synergy may be sought among the properties of the nanomaterials comprising the immunosensors.…”