INVESTIGADORES
BATTAGLINI Fernando
artículos
Título:
Recognition-Driven Layer-by-Layer Construction of Multiprotein Assemblies on Surfaces: Biomolecular Toolkit for Building Up Chemoresponsive Bioelectrochemical Interfaces
Autor/es:
D. PALLAROLA; C. VON BILDERING; L.I. PIETRASANTA; N. QUERALTO; W. KNOLL; F. BATTAGLINI; O. AZZARONI
Revista:
PHYSICAL CHEMISTRY CHEMICAL PHYSICS
Editorial:
ROYAL SOC CHEMISTRY
Referencias:
Lugar: CAMBRIDGE; Año: 2012 vol. 14 p. 11027 - 11039
ISSN:
1463-9076
Resumen:
The development of soft bioelectronic interfaces with accurate compositional and
topological control of the supramolecular architecture attracts intense interest in the
fast-growing field of bioelectronics and biosensing. The present study explores the
recognition-driven layer-by-layer assembly of glycoenzymes onto electrode surfaces.
The design of the multi-protein interfacial architecture is based on the multivalent
supramolecular carbohydratelectin interactions between redox glycoproteins and
concanavalin A (Con A) derivatives. Specifically, [Os(bpy)2Clpy]2+-tagged Con A (Os-Con A)
and native Con A were used to direct the assembly of horseradish peroxidase (HRP) and glucose
oxidase (GOx) in a stepwise topologically controlled procedure. In our designed configuration,
GOx acts as the biorecognition element to glucose stimulus, while HRP acts as the transducing
element. Surface plasmon resonance (SPR) spectroscopy and quartz crystal microbalance with
dissipation (QCM-D) results are combined to give a close representation of the protein
surface coverage and the content of water in the protein assembly. The characterization is
complemented with in situ atomic force microscopy (AFM) to give a topographical
description of the layers assemblage. Electrochemical (EC) techniques were used to
characterize the functional features of the spontaneously self-assembled biohybrid
architecture, showing that the whole system presents efficient electron transfer and
mass transport processes being able to transform micromolar glucose concentration into
electrical information. In this way the combination of the electroactive and nonelectroactive
Con A provides an efficient strategy to control the position and composition of the protein
layers via recognition-driven processes, which defines its sensitivity toward glucose.
Furthermore, the incorporation of dextran as a permeable interlayer able to bind Con A
promotes the physical separation of the biochemical and transducing processes, thus
enhancing the magnitude of the bioelectrochemical signal. We consider that these results are
relevant for the nanoconstruction of functional biointerfaces provided that our experimental
evidence reveals the possibility of locally addressing recognition, transduction and amplification
elements in interfacial ensembles via LbL recognition-driven processes.