This thesis described the modification of glassy carbon (GC) electrodes with aminophenyl (AP) films via in situ reduction of aminobenzene diazonium ions. The characterisation of the AP modified GC was conducted electrochemically by oxidation of the AP functionalities in acidic aqueous conditions. Ferricyanide and ruthenium hexamine redox probes were also used to investigate the blocking properties of the AP films. Before electrochemical oxidation of the AP functionalities, AP films were shown to have a nett positive charge at pH 7. After electrochemical oxidation in protic conditions, the film was either neutral or negatively charged. The preparation of diazonium cation terminated surface, which is termed 'sticky surface', by reaction of the AP modified electrodes with NaNO₂ in acidic condition, was investigated and the sticky surface was electrochemically characterised. More than one species was formed in the reaction of the AP film with NaNO₂. The reactions of sticky surface with aniline, citrate- and thiol-capped gold nanoparticles (Au-nps) were also studied. Spontaneous reaction of sticky surface with thiol-capped Au-nps had been achieved, and suggested that the reaction leads to the formation of Au–C bonds, via the loss of nitrogen. However, for the reaction of the sticky surface with citrate-capped Au-nps, it was unclear whether covalent bonding had been achieved. The reason for this was due to the possibility of an electrostatic interaction between the negatively charged citrate-capped Au-nps and the positively charged sticky surface. The stability of the sticky surface in acidic aqueous conditions was studied electrochemically and by reaction with thiol-capped Au-nps. It was found that the diazonium cations on the sticky surface are not stable over one hour in aqueous acidic conditions, or even in low temperature. The electro-catalytic activity of the thiol-capped Au-nps attached to the GC electrode via sticky surface towards the oxidation of ascorbic acid was briefly examined, and the surface was found to catalyse the oxidation reaction.

Protein PEGylation (covalent attachment of poly(ethylene glycol) or PEG to proteins) is an excellent example of a drug delivery system that improves pharmacokinetics and pharmacodynamic properties of therapeutics. However, although PEGylation is clinically proven and attracts both scientific and commercial interest, the technique is associated with many process constraints, in particular related to controlling the number of conjugated PEG chains. A novel, solid-phase PEGylation methodology was attempted to overcome the drawbacks of the commonly used solution-phase methods for preparing PEGylated products. The solid-phase PEGylation methodology involved conjugating protein onto a tethered PEG derivative attached onto a solid matrix, followed by hydrolytic cleavage of the PEG chain from the solid matrix under mild conditions to yield PEGylated protein in free solution. PEGs with molecular weights (MWs) 2000 and 4000 Da were used and a heterobifunctional PEG derivative, α-(β-alanine)-ω-carboxy PEG, with a cleavable β-alanine ester terminal was prepared for surface grafting and protein conjugation. The amine terminal of this PEG derivative was used for grafting PEG onto carboxy functionalized hydrophilic Sephadex and hydrophobic polystyrene derivatives. The free carboxyl terminal was used for protein conjugation via amine coupling. A kinetic study of PEG-surface grafting was performed to understand the influence of a number of parameters on the PEG surface concentration and its conformation, including temperature, reaction time, nature of the matrix, solvent and base, and MW of PEG. PEG grafted matrices were characterized using various surface characterization tools including Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). Higher PEG grafting was observed with polystyrene matrices (up to 0.3 mmol/g) than either of the Sephadex derivatives (less than 0.15 mmol/g) using both molecular weights. Detailed surface characterization using XPS studies showed a layer thickness of 11.87 nm was achieved with polystyrene matrices using 4000 Da PEG derivatives after a grafting period of 72 hours at 40°C, indicating the presence of brush conformations for the grafted PEGs. In contrast, mushroom conformations were observed for PEG molecules grafted on both carboxymethyl and carboxypentyl Sephadex derivatives after the same reaction period, with a layer thickness of 2.62 nm and 4.14 nm respectively. Optimized PEG grafting and hydrolysis conditions were developed for solid-phase protein PEGylation using Cytochrome c as a model protein. The presence of PEGylated species were detected by size exclusion chromatography (SEC) from Sephadex derivatives but were absent when using polystyrene matrices. Both Sephadex derivatives gave mainly multi-PEGylated species with poor yields, in place of the expected mono-PEGylated products. A solution-phase PEGylation using the same PEG derivatives was performed successfully and various PEGylated species were identified and characterized using SEC and gel electrophoresis, based on their viscosity radius. An examination of the surface characteristics of the PEG-grafted was carried out by XPS, showing that protein conjugation was greatly influenced by surface force interactions, which depended on the PEG grafting densities and the nature of the solid matrices. Finally, fluorescent images obtained using confocal microscope with fluorescein isothiocyanate labelled Cytochrome c provided supporting evidence regarding the factors that constrained the solid-phase PEGylation process.

Ce travail est consacré au développement d'une approche électrochimique permettant de préparer, à base de formes allotropiques différentes du carbone, divers matériaux poreux ayant comme : i) leurs éléments (feuillets de graphite, graphène, nanotubes et fullerène C60) sont attachés entre eux de façon covalente par les chaînes d'atomes du carbone de longueur différents, ii) les larges espaces internes (interstices ou les pores aléatoires) pouvant accommoder les cations, premièrement de métaux, plus grandes que Li+ (K+, Mg2+, mais aussi Alk4N+ etc.), iii) flexibilité tout en gardant la stabilité chimique des formes du carbone utilisées pour leur assemblage. Ainsi, une structure lamellaire à multicouches a été élaborée, une sorte du "graphite artificiel" avec les distances interstitielles à la demande : de 4.48 Å (avec -CH2- comme l'espaceur entre les couches) a 5.15 Å (avec -CH2C≡C-), contre 3.35 Å dans le graphite naturel. Élaborée dans ce travail, la méthode d'assemblage des matériaux stratifiés à base du graphite/graphène s'appuie sur le nouveau paradigme de greffage électrochimique aux interfaces carbonées (greffage électrophile, permettant de fixer à l'interface le linkeur CH2I ou CH2Br), et la fixation radicalaire d'une couche suivante par la réduction mono-électronique de sa liaison C-Hal. Les agents d'assemblage, Me3SiCH2I ou Me3SiC≡CCH2Br, dans un milieu universel CH3CN/0.1 M Bu4NPF6 ont été montrés être très efficaces pour ce processus. Un autre agent de couplage, 1,5-dibromopentane, a été utilisé pour assemblage cathodique (nucléophile/radicalaire) des nano-formes du carbone entre eux pour donner les matériaux poreux sans une structure spécifique régulière, les "nano-mousses", potentiellement convenable pour les anodes des batteries aux ions de Li et les batteries "post-Li-ions". Par la méthode combinée anodique/cathodique en utilisant Me3SiCH2I comme l'agent de fixation, un silsesquioxane F-@Ph8T8 avec l'ion fluorure encapsulé a été immobilisé sur le carbone vitreux ; l'interface ainsi préparée a montré son potentiel pour la détection d'exposition aux ions de Li+ ou de H+. Les structures préparées ont été étudiées par la voltampérométrie cyclique, chronocoulométrie, la spectroscopie d’impédance électrochimique et la spectroscopie électronique à balayage. / This work is dedicated to the development of an electrochemical approach to prepare, starting from different allotropic forms of carbon, various porous materials having particular features: i) their constituting elements (graphite sheets, graphene, nanotubes and fullerene C60) are covalently attached between them by the chains of carbon atoms of different length, ii) their large internal spaces (interstices or random pores) can accommodate the cations, - primarily metals, - larger than Li+ (e.g. K+, Mg2+, and also Alk4N+ etc.), iii) they are flexible while keeping the chemical stability of the carbon forms used for their assembly. Thus, a multilayered lamellar structure was developed, a kind of “artificial graphite” with interstitial distances on demand: from 4.48 Å (with -CH2- as the linker) to 5.15 Å (with -CH2C≡C-), against 3.35 Å in natural graphite. The method of assembly of graphite/graphene-based laminated materials developed in this work is based on the new electrochemical paradigm of grafting to carbon interfaces (electrophilic grafting, making possible to fix the CH2I or CH2Br functions at the interface) and the radical grafting of the subsequent layer by the mono electronic reduction of their C-Hal bond. New coupling agents, Me3SiCH2I or Me3SiC≡CCH2Br, in a universal CH3CN/0.1M Bu4NPF6 medium, have been shown to be very effective for this process. Another coupling agent, 1,5-dibromopentane, was used for cathodic (nucleophilic/radical) assembling of carbon nano forms between them to give porous materials without a regular specific structure, the “nano-sponges”, potentially suitable for making the anodes of Li-ion batteries and “post-Li-ion” batteries. Using the combined anodic/cathodic grafting method using Me3SiCH2I as the fixing agent, the fluoride encapsulating silsesquioxane F-@Ph8T8 was immobilized on glassy carbon; the thus prepared interface has shown its promise for devices detecting the exposure to Li+ or H+ ions. The prepared structures were studied by cyclic voltammetry, chronocoulometry, electro-chemical impedance spectroscopy and scanning electron spectroscopy.

En vue de vectoriser de façon ciblée des microcapsules de parfum vers un substrat textile pour des applications lessivielles, ce travail de thèse s’est consacré à la modification chimique de la surface de ces microcapsules en milieu aqueux par des polysaccharides ayant des affinités particulières pour les substrats de cellulose (agent d’aide au dépôt). Pour ce faire, une approche mettant en jeu des fonctionnalités époxy a été développée en deux étapes : fonctionnalisation de la surface des microcapsules par des molécules relais, puis greffage covalent d’un polysaccharide via la fonctionnalité époxy. Après chaque étape de greffage covalent en surface des microcapsules, différentes stratégies de caractérisations ont été mises en place (spectroscopies infrarouge, RAMAN, RMN du solide, XPS, ATG, mesure du potentiel zêta, gravimétrie, microscopie optique et fluorescente). Des expériences témoins ont également été réalisées pour prouver la non-adsorption des greffons de surface sur les microcapsules. La synthèse et le greffage de polysaccharides marqués avec des sondes fluorescente, alcyne et méthacrylate nous ont également permis d’appuyer nos conclusions. Afin d’envisager des modifications chimiques en milieu aqueux, la stabilité des composés époxy dans l’eau a dû être étudiée de façon précise par spectroscopie RMN en solution et nous avons abouti avec succès à une meilleure compréhension des phénomènes réactionnels époxy-amine et époxy-hydroxyle en milieu aqueux.Enfin, une enzyme (la lipase) a également pu être greffée de façon covalente via la fonctionnalisation époxy tout en conservant son activité catalytique. / Colloidal suspensions are of paramount significance in industrial applications. They are employed in various domains like paintings, inks, pigments, pharmacology, cosmetics, food,textile, composite materials or waste water treatment. Properties of colloids strongly depend on parameters such as the chemical composition, dimensions or morphology. To confer additional features to the colloids, i.e. stability, compatibilization, targeting, stealth properties and so on, it is also crucial to tailor their surface functionalization. In this work, we intend to develop a methodology allowing for tuning the surface properties of highly cross-linked fragrance microcapsules to graft polysaccharides. To do so, the first objective of this work is to identify functionalities at the surface (of the colloids) amenable to post-modifications. Based on this crucial insight, suitable surface chemistries are further explored to impart new properties to the colloids. Thus the presence of amine functions is highlighted by ninhydrine tests and then exploited to incorporate new functionalities at the surface of colloids.Incorporation of fluorescent tags (such as  Rhodamine Isothiocyanate, RITC), intermediate polymer epoxy chains (α,ω-epoxy functionalized polyethylene glycol or PGMA) are performed. Depending on the nature of the moieties to be grafted, the resulting colloids are subsequently characterized by Confocal Laser Scanning Microscopy (CLSM), FTIR, XPS, and RAMAN Spectroscopy. After this first step of functionnalization, epoxy rings at the surface are used to postgraft polysaccharides.

Les fonctions organiques du corps humain sont liées à des constants biologiques. Variations de ces constantes induisent divers états pathologiques. Parmi ces constantes, le pH constitue le cœur de ces travaux de thèse. Chez les êtres vivants, les fonctions biologiques dépendent de constant acides ou alcalines. En fait, l’action d’une protéine dépend du pH du milieu environnant. Une valeur inadéquate du pH rend les protéines inactives ce qui est délétère pour l’organisme. Il existe donc un besoin pour des capteurs de pH qui puissant être utilisés dans le corps humain pour des applications cliniques (échelle macroscopique), sur des cellules en culture pour des recherches en biologie (échelle mesoscopique) et pour étudier les échanges ioniques au niveau des membranes cellulaires pour des travaux plus fondamentaux (échelle microscopique). Parmi le large éventail de technologies potentiellement candidates pour ces applications, la mesure de pH par fibre optique exploitant la fluorescence permet d’être adaptée aux trois échelles dimensionnelles susnommées. Ce manuscrit de thèse adresse des contraintes par l’étude de capteurs fluorescents à fibre optique utilisant deux types d’indicateur de pH: les SNARF et la fluorescéine. En parallèle de ces développements expérimentaux, des descriptions mathématiques des propriétés de fluorescence de ces deux indicateurs sont proposées. Ces descriptions permettent de progresser vers une mesure du pH sans calibration / Organic functions of the human body are linked to biological constants. Variations of these constants induce pathological troubles. Among these constants, the pH is the central subject of this PhD work. In living beings, biological functions are related to either acid or alkaline constants. Indeed, the action of a protein depends on the surrounding pH. An inadequate value of the pH makes the proteins non active which is deleterious for the organism. There exist a need for pH sensors which can be used in the human body for clinical applications (macroscopic scale), on cells in culture for biology researches (mesoscopic scale) or at a cell membrane level for more fundamental studies (microscopic scale). Among the wide range of technologies potentially useful for these applications, fiber optic fluorescence pH sensing offers the possibility to be adapted to the three above mentioned dimensional scales. This PhD dissertation addresses these constraints by studying fluorescence fiber optics pH sensors using two kind of pH indicators: SNARF and fluorescein. Together with these experimental studies, mathematical descriptions of the fluorescence properties of these indicators are proposed. They allow progressing towards calibration free pH sensing.

Dans une thématique de plus en plus importante qui est celle du développement durable, il est aujourd’hui nécessaire d’adapter les réactions chimiques aux contraintes écologiques. Le développement de catalyseurs existe depuis le début de la chimie. Cependant, la compréhension des mécanismes mis en jeu lors des réactions chimiques est beaucoup plus récente, et ce, grâce à l’apparition de techniques d’analyse qui permettent de sonder les systèmes à différentes étapes des réactions. De nombreux catalyseurs moléculaires ont été développés, avec de très bons résultats au niveau des nombres de cycles catalytiques et de la sélectivité des réactions. Toutefois, ces catalyseurs sont souvent constitués d’un centre métallique de type iridium, ruthénium, palladium, rhodium ou platine, qui sont des métaux chers non biocompatibles. D’autre part, les oxydations sont des transformations chimiques très importantes. Des conditions de réactions dures, avec des oxydants stœchiométriques, souvent toxiques ou nocifs, sont généralement utilisées. Au contraire, des systèmes biologiques sont capables d’effectuer l’oxydation de molécules organiques en utilisant le dioxygène de l’air, en présence uniquement de protons, d’électrons, dans des conditions physiologiques. Quel grand défi pour les chimistes que d’arriver à développer des systèmes capables de mimer les systèmes biologiques, avec des catalyseurs composés de métaux biocompatibles tels que le cuivre, le manganèse et le fer. Le développement de ces catalyseurs biomimétiques demande une très bonne compréhension des systèmes biologiques, où les première et seconde sphères de coordination sont primordiales pour l'efficacité et la sélectivité des réactions. De nombreux complexes de fer(II) ont été développés comme catalyseurs dans notre équipe lors de thèses précédentes. L’interaction d’oxydants chimiques avec ces complexes a été étudiée, et une partie de mon projet a été de modifier les ligands pour augmenter la stabilité des catalyseurs, permettant l’augmentation de la sélectivité et les rendements de l’oxydation du cyclohexane et de l’anisole. Deux autres projets ont nécessité la fonctionnalisation d’un ligand utilisé communément au laboratoire pour son greffage covalent sur une électrode d’or ou dans une protéine. Afin de contrôler l’apport d’électrons au centre métallique pour réaliser l'activation réductrice du dioxygène, le complexe fonctionnalisé a été greffé sur des électrodes d’or. Le greffage sur électrode d’or a permis de mettre en avant la formation d’une monocouche homogène. Les premiers tests de réactivité de la SAM avec le dioxygène ont été également effectués. D’autre part, dans le but d’améliorer les rendements ainsi que la sélectivité des réactions en catalyse d’oxydation, un autre complexe fonctionnalisé a été greffé covalemment dans une protéine. Le greffage du complexe dans la β-lactoglobuline a permis de développer une nouvelle méthode de dosage du complexe de fer au sein de la protéine. Il a été possible de générer un intermédiaire réactionnel Feᴵᴵᴵ-peroxo, et les premiers tests en catalyse d’oxydation du thioanisole ont montré que la métalloenzyme artificielle permet d’observer une énantiosélectivité intéressante par rapport au complexe en solution. / According to sustainable development it is necessary to adapt chemical reactions to ecologic constraints. Oxidation reactions are useful transformations. Nevertheless, reaction conditions used are frequently harsh, with oxidants used in stoichiometric amount (often harmful or toxic), and lead to products formation with low selectivity. Biological systems such as metalloenzymes, are able to perform small organic molecule oxidation following O₂ activation. These reactions are achieved in physiological conditions, and with a high selectivity. Deciphering the reaction mechanism of these biological catalysts has stimulated the development of synthetic analogues such as non heme iron(II) complexes bearing amine/pyridine ligands. Reaction of these Fe(II) precursors with H₂O₂ or a single oxygen atom leads to formation of Fe(III)-OOH, Fe(III)-(O₂) and Fe(IV)=O, identified as potent oxidizing species in biological systems such as cytochromes P450. In this work, ligands were functionalized to graft iron(II) complexes on gold surface or in the β-lactoglobuline protein in order to use O₂ as oxidant or to improve yields and selectivity, respectively. Complexes grafted on gold surface were characterized by cyclic voltammetry, AFM and XPS. It has been demonstrated that it is possible to exchange exogenous ligands of the iron complex grafted on gold electrode. Preliminar reactivity tests using this grafted complex and O₂ were performed. A new artificial metalloenzyme was synthesized by covalent grafting of a functionalized iron(II) complex on β-lactoglobuline. The system was characterized, and a new method of iron(II) titration in the protein was devised. Using hydrogen peroxide, an Fe(III)-(η²-O₂) intermediate was generated and indentified in the biohybrid system, and catalytic thioanisole oxidation was observed. Interestingly, the sulfoxide product formation was shown to be enantioselective under these conditions.

The use of kaolinite for preparing functionalized materials for specialized applications is still a relatively niche research subject. This is in spite of its low cost, high availability, and the potential for covalently grafting organic functional groups to its inner and outer surfaces. These grafted compounds have been shown to be highly resistant to heat and solvents, making them very useful for certain applications, for example in polymer nanocomposite materials that require high thermal resistance during polymer processing. Solid state NMR has been shown to play an essential role in solving the structure of functionalized kaolinite materials, however the current knowledge base for these functionalized kaolinites is notably lacking for some nuclei such as 1H, 27Al and 17O. Research was undertaken to address these concerns by developing new synthetic strategies for preparing kaolinite based materials for use as nanocomposites and to examine commonly prepared modified kaolinite precursors materials by 1H and 27Al MAS NMR in an attempt to demonstrate their utility for characterizing kaolinite intercalated and grafted complexes. Solid state 1H NMR of a natural kaolinite, kGa-1b, identified two main proton signals attributed to inner and inner surface hydroxyl protons. The different affinity of these two types of hydroxyl groups towards exchange with deuterium was used to differentiate between the two. The 1H NMR spectra of a DMSO intercalated kaolinite, kDMSO, and a methanol grafted kaolinite, kmethoxy, were fitted with high accuracy using models consistent with the known structures of these materials. The 27Al MAS NMR spectra of a natural kaolinite, kGa-1b, a DMSO intercalated kaolinite, kDMSO, and a methanol grafted kaolinite, kmethoxy measured at 21.1T showed little difference between one another, while noticeable differences could be seen at 4.7T. 27Al MQMAS experiments found almost no difference between these materials in the multiple quantum dimension, suggesting the differences that were observed are a result of differences in quadrupolar parameters rather than chemical shifts. The 27Al NMR spectra of kGa-1b, kDMSO and kmethoxy were fitted with good accuracy using models consistent with known structures of these materials. Different Al(III) sites with CQ values varying by up to 0.6MHz were found. The 27Al NMR spectra of two different methanol grafted kaolinites were also compared and it was found that the intensities of the sites with lower values of CQ were dependent on the quantity of grafted aluminum sites. The interlayer space of kaolinite was functionalized with a block copolymer: poly(ethylene)-block-poly(ethylene glycol) using a kaolinite pre-intercalated with DMSO, kDMSO, and with a biodegradable polymer: poly(lactide) using a kaolinite pre-intercalated with urea, kurea, both by using melts of the polymer. The polymers were found to completely displace their precursors from the interlayer space giving a monolayer type arrangement of the polymer. Attempts were made to graft compounds containing polymerizable functional groups: 3-allyloxy-1,2-propanediol and ethylene glycol vinyl ether to kaolinite’s inner surfaces using a kaolinite pre-intercalated and grafted with methanol, kmethoxy, and a kaolinite pre-intercalated with DMSO, kDMSO, respectively. Both compounds were found to displace their precursors from the interlayer space, adopting a monolayer type arrangement. 13C and 29Si NMR results suggest 3-allyloxy-1,2-propanediol’s allyl group remains intact and partially keys into the clay mineral’s siloxane rings. Ethylene glycol vinyl ether was found to undergo intramolecular cyclization to form an acetal product, consuming its vinyl group in the process. This reaction was observed using an unmodified kaolinite, kGa-1b, suggesting that the clay mineral’s surfaces, both inner and outer, act as an acid catalyst.