Nanobiotechnology aims to exploit biomolecular recognition and self-assembly capabilities for integrating advanced materials into medicine and biology. However frequent problems are encountered at the interface of substrate-biological molecule, as the direct physical adsorption of biological molecules is dependent of unpredictable non-specific interactions with the surface, often causing their denaturation. Therefore, a proper functionalization of the substrate should avoid a loss of biological activity. In this work we address the functionalization of the semiconductor GaN (0001) for biosensing applications. The basic interest of using III-V class semiconductors is their good light emitting properties and a fair chemical stability that allows various applications of these materials. The technology chosen to elaborate GaN-specific peptides is the combinatorial phage-display method, a biological screening procedure based on affinity selection. An M13 bacteriophage library has been used to screen 1010 different peptides against the GaN (0001) surface to finally isolate one specific peptide. The preferential attachment of the biotinylated selected peptide onto the GaN (0001), in close proximity to a surface of different chemical and structural composition has been demonstrated by fluorescence microscopy. Further physicochemical studies have been initiated to evaluate the semiconductor-peptide interface and understand the details in the specific recognition of peptides for semiconductor substrates. Fourier Transform Infrared spectroscopy in Attenuated Total Reflection mode (FTIR-ATR) has been employed to prove the presence of peptides on the surface. Our Atomic Force Microscopy (AFM) studies on the morphology of the GaN surface after functionalization revealed a total surface coverage by a very thin, homogeneous peptide layer. Due to its good biocompatibility, functionalized GaN devices might evolve in a new class of implantable biosensors for medical applications.

Bacteriorhodopsin (BR) is a robust trans-membrane protein that functions as a light-driven proton pump, thus is an excellent candidate for biophotonics applications. For the development of new optical devices, the buildup of stable BR matrices has to be optimised. In this work, we present a multi- technique approach: the combination of optical waveguide lightmode spectroscopy (OWLS), atomic force microscopy (AFM) and multi-photon microscopy (MPM) aiming to analyze the optical and physico-chemical properties of BR embedded in polyelectrolyte multilayers (PEM) in its membrane bound form (purple membrane, PM), as well as solubilized BR immobilized within a photonic structure built of porous silicon (PSi). OWLS measurements revealed the possibility of incorporation of PM-BR layers into PEmultilayers. The calculated thickness and refractive index of the adsorbed layers demonstrate the successful adsorption of PM on top of the positively or negatively charged PE layers. Morphological studies by AFM proved a complete coverage of the positively charged PE layer with PM patches. As for the other model system, photonic responses of BR, after being immobilized within PSi substrates, have been evaluated using multi-photon microscopy. Fluorescence emission and second harmonic generation (SHG) of the BR-PSi system were observed at some particular pores of PSi and subsequent enhancement of the signal arising from the BR adsorbed within the pores was detected. Our results constitute the first steps of two interesting and innovative biomimetic approaches for the future design and development of BR based integrated optical devices.

The membrane protein bacteriorhodopsin in its native membrane bound form (purple membrane) was adsorbed and incorporated into polyelectrolyte multilayered films, and adsorption was in situ monitored by optical waveguide light- mode spectroscopy. The formation of a single layer or a double layer of purple membranes was observed when adsorbed on negatively or positively charged surfaces, respectively. The purple membrane patches adsorbed on the polyelectrolyte multilayers were also evidenced by atomic force microscopy images. The driving forces of the adsorption process were evaluated by varying the ionic strength of the solution as well as the purple membrane concentration. At high purple membrane concentration, interpenetrating polyelectrolyte loops might provide new binding sites for the adsorption of a second layer of purple membranes, whereas at lower concentrations only a single layer is formed. Negative surfaces do not promote a second protein layer adsorption. Driving forces other than just electrostatic ones, such as hydrophobic forces, should play a role in the polyelectrolyte/purple membrane layering. The subtle interplay of all these factors determines the formation of the polyelectrolyte/purple membrane matrix with a presumably high degree of orientation for the incorporated purple membranes, with their cytoplasmic, or extracellular side toward the bulk on negatively or positively charged polyelectrolyte, respectively. The structural stability of bacteriorhodopsin during adsorption onto the surface and incorporation into the polyelectrolyte multilayers was investigated by Fourier transform infrared spectro-scopy in attenuated total reflection mode. Adsorption and incorporation of purple membranes within polyelectrolyte multilayers does not disturb the conformational majority of membrane-embedded R-helix structures of the protein, but may slightly alter the structure of the extramembraneous segments or their interaction with the environment. This high stability is different from the lower stability of the predominantly β-sheet structures of numerous globular proteins when adsorbed onto surfaces.