A modificação de eletrodos por sistemas auto-organizados (SAM) é um dos temas mais discutidos em eletroquímica e eletroanalítica, principalmente por elevar a biocompatibilidade destas superfícies. O foco principal deste trabalho está na transferência eletrônica através das SAMs de moléculas de alcanotióis sobre a superfície de eletrodos de ouro. Com a finalidade de investigar o comportamento eletroquímico do par redox [Fe(CN)6]3-/[Fe(CN)6]4-,, as superfícies de ouro foram modificadas com SAMs de alcanotióis com cadeias carbônicas de diferentes comprimentos e suas misturas. Desta forma, foi possível observar que as superfícies modificadas com monocamadas com 2, 3 ou 4 átomos de carbono em sua cadeia permitem que a transferência de elétrons ocorra através da SAM, produzindo perfis voltamétricos muito similares aos observados com a superfície de ouro sem a modificação. Com cadeias carbônicas longas (até 11 átomos de carbono), a transferência eletrônica é totalmente bloqueada e nenhum perfil voltamétrico foi observado. Com o objetivo de explorar o bloqueio total da superfície modificada de moléculas de alcanotióis de cadeias longas, os excessos superficiais foram calculados por meio da carga voltamétrica obtida da dessorção redutiva. Observaram-se valores de excesso superficial próximos a 9,0x10-10 mol/cm2, o que demonstra uma cobertura completa dos sítios ativos da superfície, justificando assim a ausência da resposta eletroquímica. A superfície modificada com as misturas de monocamadas de cadeias carbônicas longas e curtas promoveu respostas voltamétricas interessantes, desde que as moléculas de cadeias curtas contenham menos que 4 átomos de carbono. Neste caso, respostas voltamétricas sigmoidais similares às de ultramicroeletrodos (UMEs) foram obtidas. Este efeito foi associado com a formação de conjuntos de UMEs formados na superfície do eletrodo. O raio e a separação entre os UMEs nestas superfícies foram calculados como sendo 42 e 221 µm, respectivamente. Um total de 60 UMEs foi encontrado na superfície modificada. Estes UMEs foram formados por moléculas de cadeias menores totalmente emparedadas por cadeias maiores (moléculas de ácido 11- mercaptoundecanóico). Este comportamento reforça o propósito de que a transferência dos elétrons ocorra através das monocamadas de cadeia menor e que as moléculas de cadeia longa bloqueiam a superfície. Todos os experimentos voltamétricos foram seguidos de análises de espectroscopia de impedância eletroquímica os quais permitiram determinar a resistência de transferência de carga para cada modificação. Superfícies modificadas com alcanotióis de cadeia longa apresentaram elevados valores de transferência de carga, enquanto que as superfícies modificadas com alcanotióis de cadeia curta apresentam baixos valores de transferência eletrônica, em concordância com o comportamento voltamétrico observado. Finalmente, a organização de cada monocamada foi monitorada utilizando a microbalança eletroquímica de cristal de quartzo. Pela variação de massa com o tempo de imersão do eletrodo na solução contendo o alcanotiól de interesse foi possível observar etapas de adsorção/dessorção, com o aumento da organização da monocamada. Os valores teóricos de variação de massa calculados para a formação de uma monocamada completa na superfície eletroquímica, apresentaram excelente concordância com os dados experimentais obtidos pelo monitoramento do sistema ao longo dos experimentos (depois de aproximadamente 2 horas de imersão). / The modification of electrode surfaces with self-assembled monolayer (SAM) is one of the top subjects in electrochemistry and electroanalysis, mainly due to its much enhanced biocompatibility. In this study, the focuses are centered in the electron transfer through SAMs of alkanothiols assembled onto gold electrode surfaces. In order to investigate the electrochemical behavior of the redox couple [Fe(CN)6]3-/[Fe(CN)6]4-, Au surfaces were modified with SAMs of long and short carbon chains alkanethiols, as well as with their mixtures. In this way, it was possible to observe that surfaces modified with monolayers of 2, 3 or 4 carbon atoms in the chain allowed electrons transfer to occur through the SAM, generating a voltammetric profile very similar to those observed on bare Au surfaces. With longer carbon chains (up to 11 carbon atoms), the electron transfer was totally blocked and none voltammetric profile was observed. Aiming to explain the total blockage of the surface with long chain alkanethiols modifications, the surface excess of molecules were measured by the voltammetric charge required for their reductive desorption. It was always observed surface excesses near to 9.0x10-10 mol/cm2, which is related to a full coverage of the active sites on the surface, thus justifying the absence of electrochemical response. The surface modification with mixtures of long and short carbon chains monolayers promoted very interesting voltammetric responses, as long as the short chains contain less than 4 carbon atoms. In this case, sigmoidal voltammetric responses were obtained, very similar to those observed for ultramicroelectrodes (UMEs). Such effect was associated to the formation of UMEs arrays in the electrode surface. The radius and separation of each UME in this surface array was evaluated as 42 and 221 µm, respectively. A value of 60 UMEs were found for the entire surface. These UMEs were formed by the short chain molecules totally walled by the long chain ones (mercaptoundecanoic acid molecules). Such behavior reinforces the proposition of electron transference through the short chain alkanethiol layer and surface blockage by the long chain one. All the voltammetric experiments were followed by electrochemical impedance spectroscopic analysis which allowed were determined the charge transfer resistance for each modification. Very large values were determined for long carbon chains while for the short chains much smaller resistance values were found, as expected by the voltammetric behavior. Finally, the assembling of each monolayer was followed by quartz crystal microbalance measurements. By the electrode mass variation with time of immersion in a solution containing the alkanethiol of interest it was possible to observe several steps of adsorption/desorption, as the organization of the monolayer increases. The theoretical values for mass variation for a full monolayer in the electrochemical surfaces were found to be in highly agreement with the experimental ones, measured in long time experiments (after approximately 2 hours of immersion).

Este trabalho descreve a preparação, a caracterização e o uso de um biossensor bienzimático confeccionado com as enzimas glicose oxidase e frutose dehidrogenase imobilizadas em camadas auto-organizadas ou self-assembled monolayers (SAMs) de cistamina para a quantificação de açúcares em alimentos. Após o preparo do eletrodo de ouro com a SAM de cistamina, biossensores foram construídos e para obtenção de melhores respostas, condições foram otimizadas, tais como: concentração do mediador tetratiafulvaleno (TTF), porcentagem de glutaraldeído, temperatura e tempo de vida do biossensor. Com as condições estabelecidas, fez-se então, a determinação analítica da D-glicose e da D-frutose em eletrólito puro pelo método da adição de padrão e os resultados foram obtidos por voltametria cíclica e cronoamperometria. A corrente de pico de oxidação do mediador de elétrons (TTF) aumentou proporcionalmente com o aumento da concentração e não ocorreram deslocamentos nos potenciais de pico. Os limites de detecção (LD) foram encontrados por meio do desvio padrão da média aritmética de dez amperogramas do branco no potencial equivalente aos dos picos de oxidação do mediador de elétrons TTF, juntamente com o valor do coeficiente angular da curva analítica. Após a obtenção da curva analítica o biossensor foi aplicado diretamente em amostras de refrigerante dietético e não dietético, bem como em amostras de adoçantes comerciais, onde foram realizados testes comparativos da resposta dos biossensores. Para o eletrólito puro e amostras de refrigerante dietético e de adoçantes, ou seja, onde não há presença de D-glicose e D-frutose, notou-se a ausência de corrente de pico, enquanto que para as amostras de refrigerante não dietético, houve um valor significativo de resposta de corrente, indicando a presença dos açúcares em estudo. Com o propósito de verificar a influência de interferentes e o efeito de matriz, foi construída uma curva analítica para a D-glicose e para a D-frutose, em amostras de refrigerante dietético e adoçantes, onde foram obtidas as menores quantidade destes açúcares. Para o refrigerante não dietético, foi determinado o valor inicial dos açúcares presentes nas amostras. Pode-se afirmar que a utilização dos biossensores baseados nas enzimas GOx e FDH mostraram-se eficientes para a determinação dos açúcares D-glicose e D-frutose nas amostras analisadas (com diferença significativa nos valores de corrente), apresentando uma resposta rápida, além da eliminação do efeito da matriz. A utilização do mediador de elétrons (TTF) possibilitou a reação em potencial próximo de zero, diminuindo o efeito de interferentes e evitando a desnaturação das enzimas. / This work describes the preparation, characterization and application of a bienzymatic biosensor based in the glucose oxidaze and fructose dehydrogenase immobilized on self-assembled monolayer of cystamine for sugar quantification in foodstuff. After the modification of the gold electrode with cystamine, the biosensors were developed and optimized for best responses. Optimization parameters were: mediator tetrathiafulvalene (TTF) concentration, glutaraldehyde percentage, temperature and life time of the biosensor. With the best conditions established, the analytical determinations of d-glucose and d-fructose in pure phosphate buffer were conducted by the standard additions method and the results obtained by cyclic voltammetry and chronoamperometry. The oxidation peak current related to the TTF voltammetric behavior raised proportionally to the increasing concentration of d-glucose or d-fructose, in a given and constant peak potential. The methodology detection limits were found using the standard deviation of ten chronoamperograms of the blank solution, in the potential value corresponding to that of TTF oxidation, and the slope of the analytical curve. After the analytical curve acquirement the biosensor was directly applied in samples of diet or non-diet softdrinks, as well as in commercial sweeteners samples, with comparative tests of the biosensor responses. For pure electrolyte and for diet foodstuff samples, i.e., were there is not expectation for d-glucose or d-fructose existence, it was detected the lack of the voltammetric peak associated with the mediator oxidation. In non-diet samples, a pronounced voltammetric peak was obtained, testifying the presence of sugar in the electrolytes under study. The matrix effect was verified by means of an analytical curve obtained for both analytes (d-glucose and d-fructose), in diet and sweeteners samples, properly spiked with known amounts of each analyte. It can be concluded that the utilization of the biosensor based in GOx and FDH showed to be efficient for d-glucose and d-fructose determinations in the analysed samples, with a fast response time and elimination of the matrix effect. The mediator promoted the electrochemical reaction to occur in potentials very close to zero, minimizing the interferences or the enzyme denaturation.

Monocamadas auto-organizadas formadas pela quimissorção de alcanotióis sobre ouro apresentam estruturas bem definidas, organizadas e reprodutíveis. As propriedades de SAMs, aliadas à facilidade de síntese, têm atraído o interesse da comunidade cientifica o que ocasionou um grande avanço na área de nanotecnologia, especialmente em engenharia de superfícies. Neste trabalho, avaliou-se a modificação de um substrato de ouro com uma mistura dos ácidos 3-mercaptopropiônico e 11-mercaptoundecanóico. Com isso procurou-se controlar a configuração da superfície da SAM a fim de se obter um arranjo com ilhas de 3amp circundadas por 11amu, similar a um conjunto de ultramicroeletrodos. A SAM mista foi produzida por incubação de um substrato de ouro em solução etanólica contendo ambos os tiois. O estudo do processo redox do par [Fe(CN)6]4-/[Fe(CN)6]3- demonstrou que monocamadas de 3amp se comportam como a superfície de ouro não modificado, apresentando os mesmos valores de corrente de pico (Ip), potencial de pico (Ep) e resistência de transferência de carga (Rtc), pois permitem que ocorra a transferência eletrônica por efeito de tunelamento quântico dos elétrons através da monocamada. Entretanto monocamadas de 11amu demonstram comportamento isolante, apresentando uma Rtc 250 vezes maior e Ip significativamente menores que a observada para a SAM-3amp. As cargas de dessorção redutiva e variação de massa obtidas com a MECQ para a SAM mista evidenciaram alto recobrimento da superfície e um mecanismo de adsorção que varia com o tempo de incubação do substrato em solução contendo os tiois. Assim, a configuração da superfície com domínios de 3amp bem estabelecidos é alcançada com 20 horas de incubação do substrato. O perfil observado com a voltametria cíclica, aliado aos dados obtidos com a EIE comprova a existência de segregação de fases na SAM mista contendo regiões recobertas pelo 3amp (raio médio de 4,3 µm) circundadas por 11amu (com separação média de 75,42 µm). O estudo do comportamento eletroquímico do fisetin confirma o surgimento de propriedades diferenciadas com a SAM mista, onde observou-se apenas um processo redox sem que o analito permanecesse adsorvido na superfície da SAM. Todavia para a SAM-3amp e ouro não modificado foi observado dois processos redoxes seguido de adsorção irreversível do produto da reação na superfície eletródica. Utilizando a SAM mista como sensor eletroquímico para determinação de fisetin em água alcançou-se um limite de detecção de 1,67 x10-8 mol L-1. / Self-assembled monolayers obtained by chemiosorption of alkanethiols on gold present well-defined, organized and reproducible structures. The unique properties exhibited by such surfaces, allied to the great facility to obtain, have attracted the interest of the scientific community and caused a significant advance in the nanotechnology research, especially in the surface engineering one. In this work, the modification of gold substrate with a mixture of 3-mercaptopropionic acid and 11-mercaptoundecanoic acid was evaluated. The aim was control the surface of the SAMs in order to obtain an island arrangement of 3mpa isolated by 11mua, similar to an array of ultramicroelectrodes. The mixed-SAM was built by the incubation of Au substrate in an ethanolic solution containing both thiols. The redox couple [Fe(CN)6]4-/[Fe(CN)6]3- study demonstrated that pure 3mpa monolayers behaves like the bare gold surface, presenting barely the same parameters of peak current (Ip), peak potential (Ep) and charge transfer resistance (Rct), since it allows the electronic transference to occurs by quantum tunneling effect through the monolayer. On the other hand the 11mua monolayers showed an insulating behavior and a Rct value nearly 250 times greater and, consequently, Ip values significantly less than that for 3mpa. The reductive desorption charges and mass changes obtained with an EQCM for the mixed-SAM indicated the high coverage of gold surface and a adsorption mechanism that depends on the incubation time of the substrate in the thiols solution. The surface configuration with 3mpa domains is reached after 20 hours of immersion. The cyclic voltammetric profile, together with the data from electrochemical impedance spectroscopy , proved the existence of phases segregation in the mixed-SAM with surface regions covered solely by 3mpa (with mean radius of 4.3 µm) surrounded by 11mua with a mean separation of 75.42 µm. The electrochemical behavior of the flavonoid fisetin confirms the rising of differentiated properties with the mixed-SAM, where only one electrochemical process was able to be observed, without irreversible adsorption of the analyte. For 3mpa and bare gold surfaces, two (or more) electrochemical processes were observed together with the blocking of the electrode surface by irreversible adsorption of the reaction product. The utilization of the mixed-SAM with electrochemical sensor for fisetin determination in pure water yielded a detection limit of 1.67 x10-8 mol L-1.

Neste projeto estudou-se a interação da enzima peroxidase de raiz forte (HRP) em interfaces nanoestruturadas e sua possível aplicação em biossensores de peróxido de hidrogênio. Foram utilizadas as técnicas de Langmuir, Langmuir-Blodgett (LB) e automontagem por adsorção física para formar filmes nanoestruturados. A interação da enzima com espécies em interfaces foi investigada com materiais que serviram de matrizes de adsorção, ou seja, a quitosana (Ch) e o fosfolipídio 1,2-dipalmitoil-sn-glicero-3-[fosfatidil-rac-(1-glicerol)] (sal de sódio) (DPPG). Os filmes de Langmuir foram caracterizados com medidas de pressão e potencial de superfície, espectroscopia no infravermelho, e tensão superficial dinâmica. Para os filmes LB e automontados, empregaram-se espectroscopias de fluorescência, ultravioleta-visível e infravermelho e microgravimetria por cristal de quartzo. A peroxidase de raiz forte apresentou forte interação com DPPG, confirmada em filmes de Langmuir por medidas de pressão de superfície, elasticidade dinâmica e de espectroscopia de reflexão e absorção no infravermelho, com modulação por polarização (PM-IRRAS). A massa de peroxidase transferida em filmes Langmuir-Blodgett (LB) mistos com DPPG foi de aproximadamente 200 ng, de acordo com medidas com uma microbalança de cristal de quartzo. A atividade da HRP foi mantida no filme LB, inclusive com atividade catalítica maior do que em meio homogêneo e nos filmes automontados com quitosana. As medidas de atividade não afetaram a morfologia dos filmes LB, estudada com microscopia de força atômica (AFM), ao contrário dos filmes automontados. Conclui-se que a imobilização de HRP é mais eficiente num filme LB, com matriz fosfolipídica, apresentando boas perspectivas de emprego em biossensores de peróxido de hidrogênio. / A study has been performed on the interaction of the enzyme horseradish peroxidase (HRP) in nanostructured interfaces and their possible application in biosensors for hydrogen peroxide. The nanostructured films were obtained with the Langmuir, Langmuir-Blodgett (LB) and layer-by-layer (LbL) methods. The interaction between HRP and species at interfaces was investigated using materials that served as matrix for immobilization, viz. chitosan (Ch) and the phospholipid 1,2-dipalmytoil-sn-glycero-3-[phosphatidyl-rac-(1-glycerol)] (sodium salt) (DPPG). The Langmuir films were characterized with surface pressure, surface potential, elasticity measurements and polarization-modulation reflection and absorption infrared spectroscopy (PM-IRRAS). For LB and LbL films, use was made of fluorescence, absorption in the UV-vis. and infrared spectroscopy. HRP displayed strong interaction with DPPG, which was confirmed in Langmuir films with measurements of surface pressure, dynamic elasticity and PM-IRRAS. The mass of HRP transferred onto a solid support in a mixed LB film with DPPG was 200 ng, according to data from a quartz crystal microbalance. The HRP activity was preserved in the mixed LB film, with a catalytic activity that was even higher than in solution or in LbL films of HRP/Ch. The catalytic activity measurements did not affect the morphology of the LB films, studied with atomic force microscopy (AFM), in contrast to the LbL films. The main conclusion is that HRP immobilization is more efficient in an LB film with a phospholipid matrix, with good prospects for developing biosensors for hydrogen peroxide.

Eletrodos de ouro foram utilizados para preparação de eletrodos modificados com monocamadas auto arranjadas de tióis. A limpeza do substrato metálico é fundamental para que a arquitetura molecular superficial possa ser efetuada com boa estabilidade e reprodutibilidade, além de fornecer dados utilizados no cálculo de área ativa, necessários na normalização dos valores de recobrimento superficial obtidos nas determinações de dessorção do agente modificador interno, o ácido 3-mercaptopropiônico. Os eletrodos modificados consistiram no recobrimento de transdutores de ouro com ácido 3-mercaptopropiônico através da imersão do eletrodo de Au em solução 25 mmolL-1 deste composto e, em seguida, com moléculas de cisteína, através da imersão do eletrodo de Au/3-MPA em solução 0,1 molL-1 deste composto, originando um sensor do tipo Au/3-MPA/CSH. As moléculas de cisteína foram utilizadas como agente redutor para obtenção de nanopartículas de Au na superfície do eletrodo modificado, através da aplicação de 20 µL de solução de HAuCl4. Após a confirmação da ausência do par tiólico superficial responsável pela redução das nanopartículas, o eletrodo Au/3-MPA/CSH/AuNp foi utilizado na determinação de peróxido de hidrogênio em soluções de concentrações crescentes em tampão fosfato 0,1 molL-1 pH 7,2. / Gold electrodes were used for surface modification through the incorporation of self-assembled monolayers of thiols. Pre-treatment of the metal substrate is essential for achieving good stability and reproducibility of the molecular architecture in the surface developed, and provides data used in the calculation of the active area, used to normalize surface coating values, which were obtained throughout the desorption measurements of 3-mercaptopropionic acid. The modification consisted on coating gold probes with 3-mercaptopropionic acid by dipping the Au electrode in 25 mmolL-1 solution of this compound, in which were, later, anchored cysteine molecules, by dipping of the Au/3-MPA electrode in 0.1 molL-1 solution of this compound, resulting in a Au/3-MPA/CSH sensor. Cysteine molecules were used as a reducing agent to obtain gold nanoparticles on the surface of the modified electrode through application of 20 µL of HAuCl4. After confirming the absence of the thiol redox couple responsible for the reduction of gold nanoparticles, the Au/3-MPA/CSH/AuNp electrode was used to determine hydrogen peroxide in phosphate buffer 0.1 molL-1 pH 7.2.

III-V semiconductors have higher charge carrier mobilities than silicon and are used in photovoltaic devices, optical sensors, and emitters. The high injection velocities obtained with III-V channels allow for faster transistors with low power consumption. However, the large-scale implementation in electronic devices is currently limited by the defective interface formed between III-Vs and their oxides. Clean III-V surfaces are highly reactive in air and form amorphous oxides that lead to a high density of dangling bonds. Satisfying these dangling bonds has been associated with an improvement in electrical performance, directing the development of strategies that decrease the surface reactivity (chemical passivation) and the density of surface states that cause power dissipation (electrical passivation). Sulfur bonds easily to III-V surfaces and has been used to chemically and electrically passivate GaAs. In this work, we investigated liquid phase sulfur chemistries in the chemical passivation of clean InSb(100) and In0.53Ga0.47As(100) surfaces terminated by their group V elements. Our strategy consisted of maximizing the number of bonds between sulfur and antimony or arsenic. A long alkane chain thiol, 1-eicosanethiol (ET, 20 carbon atoms), was used to produce a hydrophobic surface and deposit a dense organic layer by taking advantage of the van der Waals interactions between thiol molecules. The first part of the study involved the optimization of the thiol deposition process on InSb. Self-assembled alkanethiol monolayers were formed by immersing clean InSb substrates in ET solutions in ethanol for 20 h. The layers prevented the formation of detectable oxides for 20 min based on the O Auger x-ray photoelectron spectroscopy (XPS) peak. The thiol layer was completely removed by heating the surface to 227 C in vacuum. In the second part of the study, a 20 h ET deposition was performed on In0.53Ga0.47As(100), and re-oxidation was prevented for up to 4 min based on the O 1s XPS peak. The alkanethiolate layer was removed by heating the samples to 350 C in vacuum. The sulfur coverage after 20 min and 20 h ET depositions was increased by performing a second immersion in (NH4)2S without modifying the thickness of the layer. The best process studied consisted of a 20 h immersion in ET solution followed by a short (NH4)2S step, preventing the formation of oxides for up to 9 min. This is due to the presence of available surface sites and weakly bonded molecules in the layer after a long 20 h ET process. The chemical passivation effect is not uniquely influenced by surface termination, roughness, or lattice constant, but is rather a result of a combination of these factors. Future work will involve the fabrication and electrical characterization of III-V devices modified with various chemical passivation strategies.

Le sujet de cette thèse concerne la spintronique moléculaire. Des jonctions tunnel magnétiques formées par une barrière tunnel moléculaire (monocouche auto-assemblée) insérée entre deux électrodes métalliques ferromagnétiques ont été étudiées. Afin de fabriquer les dispositifs, un procédé de greffage des molécules sur des substrats ferromagnétiques a été mis en place et une technique de lithographie a été développée pour définir des jonctions de taille submicronique. L’ensemble de ce travail expérimental a permis l’obtention de jonctions non court-circuitées et mesurables, où le transport électronique est bien du transport par effet tunnel.Les mesures de magnétotransport de ces échantillons ont amené des résultats intéressants et nouveaux : les jonctions, dont la barrière est formée par des alcanes-thiols, présentent de la magnétorésistance à température ambiante, allant jusqu’à 12%. Une partie de la thèse s’attelle à la compréhension des différents comportements magnétorésistifs observés : un modèle de barrière tunnel à deux niveaux est proposé pour les décrire.La dernière partie du travail présente des résultats préliminaires obtenus lorsque la barrière moléculaire est formée par des molécules aromatiques ou commutables et met en évidence des phénomènes nouveaux par rapport au cas précédent.L’ensemble des résultats prouve le fonctionnement de jonctions tunnel magnétiques à base de monocouches moléculaires à température ambiante et ouvre la voie à l’utilisation de molécules plus complexes pour une électronique de spin moléculaire multifonctionnelle. / This thesis work enters within the molecular spintronic fields. Magnetic tunnel junctions based on molecular self assembled monolayers have been investigated. The devices structure is a molecular monolayer inserted between two ferromagnetic electrodes.A process to graft molecules on a ferromagnet’s surface and a lithography technique have been developed to define the junctions. This experimental work has led to non short-circuited and measurable junctions, in which an electronic tunnel transport has been demonstrated.Interesting and new results have been found out from magnetoresistance measurement of the samples: junctions made with alkanes-thiols barrier have shown magnetoresistance signal at room temperature (up to 12%). In order to explain the magnetoresistive behaviour, a simple model where the barrier is discribed by two levels has been proposed.The last part of the thesis reports preliminary results obtained when the barrier is made of aromatic molecules or switchable molecules and it points out new phenomenons compared to the alkanes case.The overall work proves that devices made from magnetic tunnel junctions with self-assembled monolayers work at room temperature. It is then possible to consider switchable molecules to build multifunctional molecular spintronics devices.

L’objection générale de ces travaux est de comprendre les mécanismes d’interaction de molécules d’intérêt avec les membranes afin de faciliter la synthèse de molécules plus efficaces contre leurs cibles tout en étant moins toxique pour l’Homme. Dans la première partie de ces travaux, nous avons étudié les interactions entre ces modèles membranaires et les polyènes antifongiques, connus pour interagir avec les stérols des membranes plasmiques. Nous nous sommes particulièrement intéressés à la Nystatine et à l’Amphotéricine B, deux molécules de structure chimique très proche et actuellement utilisées dans l’industrie pharmaceutique. L’utilisation de différents modèles membranaires et de techniques adaptées a montré que la PhosphatidylEthanolamine avait très vraisemblablement un rôle primordial dans le mécanisme d’interaction de ces molécules avec les membranes. Dans la deuxième partie de ces travaux, nous nous sommes intéressés à l’inhibition de la formation du cristal d’hémozoïne formé lors de la croissance du parasite responsable du paludisme. Ce cristal est formé d’hématine, hautement toxique pour le parasite. L’hématine et l’inhibition de la formation de l’hémozoïne constituent une cible moléculaire idéale pour combattre cette maladie. La chloroquine, la méfoquine et de nouveaux inhibiteurs dérivés de la méfloquine ont été utilisés. L’étude de l’inhibition de la formation du cristal s’est faite en utilisant des monocouches de Langmuir, servant ainsi de biocapteurs. Ces travaux ont montré que l’énantiomérie, mais aussi la lipophilicité des nouveaux composés antipaludiques sont des paramètres importants en vue de la synthèse de molécules plus efficaces. / The main purpose of this work is to better understand the mechanisms of interaction between pharmaceutical relevant molecules and model membranes in order to facilitate the synthesis of new molecules, more efficient against their molecular target and less toxic for Humans. In the first part, we studied the interactions occuring between these models and antifungal polyene molecules. It has been reported that these molecules interacted preferentially with sterols. We specifically focused on Nystatin and Amphotericin B, two polyenes with a very similar chemical structure and presently used as a treatment against fungi and molds. Using different kind of model membranes, we showed PhosphatidylEthanolamine could have a very important role in the mechanism of action of these molecules. In the second part of this work, we studied the inhibition of the formation of a cristal called « hemozoïn », which is growing during the life cycle of the parasite responsible of malaria. This cristal is made of hematin, a toxic by-product of the degradation of hemoglobin, the main source of amino-acids for the parasite. Hematin and the inhibition of the growth of this cristal is a ideal molecular target to combat malaria. Chloroquine, mefloquine and new mefloquine-derivatives were studied. The study of the inhibition of the formation of the cristal was done using Langmuir monolayers as a biosensor. We showed that stereochemistry, but also lipophilicity of these compounds, are important parameters for the synthesis of more efficient antimalarial molecules.

Microarrays on gold have been used to develop new methodologies for biocatalysis and proteomic applications. The technology applies the logic of solid phase supported chemistry using self-assembled monolayers (SAMs) on a gold chip. The advantages of this technology are: i) an easy to handle platform, ii) parallel screening (64 reactions at once), iii) microliter scale reactions (1µL per sample), iv the use of mild conditions (buffers and t=37 °C), v) the absence of purification steps (only chip washing is required), vi) quick and accurate analysis by MALDI ToF MS.The metallopeptidase thermolysin was studied in peptide-coupling reactions to profile reactivity and specificity (Chapter 2). Reactivity was further investigated in transpeptidation reactions. Comparing the serine peptidase chymotrypsin with the zinc dependent thermolysin, it was found that transpeptidations proceed via N-transacylation reactions independent of a specific enzymatic catalytic mechanism (Chapter 3). These transacylations may be exploited for modifications of biocompatible and selective surfaces in ‘bottom-up’ bionanofabrication technologies. Selected peptidases with different catalytic mechanism were also arrayed to investigate polymerisation ability of dipeptides (Chapter 4). It was shown that oligomerisation can be obtained under mild conditions and a set of peptides was synthesised. Chapter 5 describes a new chemical methodology by which crude tryptic peptide digests can be trapped on chip and analysed by MALDI ToF MS without further purification steps. This dramatically improves time and cost efficiency.Finally, a new stepwise native chemical ligation methodology is proposed for amino acids, and peptides containing N-terminal cysteine residues (Chapter 6).

Explorou-se a caracterização de monocamadas e filmes LB de polianilina (PANi), e um oligômero de polianilina, chamado aqui de 16-mero, com estearato de cádmio (CdSt). A análise dos filmes de Langmuir indicou que possivelmente não há mistura em nível molecular da PANi e do 16-mero com CdSt, ainda que a quantidade adicionada em solução de polímero ou oligômero influencie a estabilidade das monocamadas. Utilizou-se UV-vis, FTIR, difração de raios-X, condutividade elétrica, elipsometria, microscopia óptica e potencial de superficie para caracterização dos filmes LB. Os resultados de UV-vis indicaram uma transferência uniforme e ainda que os filmes recém depositados encontram-se desdopados. A análise de FTIR confirma a presença de CdSt nos filmes LB, e o efeito de desdopagem em subfases neutras, corroborando os resultados de UV-vis. A difração de raios-X indica a presença de domínios separados de PANi (e 16-mero) com CdSt nos filmes LB. Resultados elipsométricos indicaram uma espessura por camada em tomo de 25 &#197. A excelente uniformidade obtida nos filmes LB mistos foi comprovada pelos resultados de microscopia óptica e potencial de superficie. Esses filmes mistos apresentaram valores de condutividade elétrica em tomo de 10-4 a 10-5 S.cm-1 (van der Pauw), tanto para a PANi quanto para o 16-mero. Investigou-se também o efeito da exposição dos filmes mistos PANi/CdSt aos raios-X. Nota-se um deslocamento para a região do vermelho, na região do visível, nos espectros de UVvis, similar à observada pela dopagem através de ácidos inorgânicos. Verificou-se que os efeitos de umidade da atmosfera de medida são predominantes nesse processo de dopagem. É feita uma análise dos resultados de condutividade elétrica, comparando-os com os encontrados na literatura. / A study has been made of composite Langmuir and Langmuir-Blodgett (LB) films of polyaniline (PANi), and a polyaniline oligomer (l6-mer polyaniline), with cadmium stearate (CdSt). The monolayers studies pointed to a phase separated system containing the polymer (or the 16-mer) and CdSt, with no mixing at the molecular leveI, although the relative contents of PANi and 16-mer in the solution have a strong influence on the monolayer stability. UV-vis, FTIR, X-ray difIraction (XRD), electrical conductivity, ellipsometry, optical microscopy and surface potential measurements were used in the LB film characterization. UV-vis results have shown a uniform transfer, with the as deposited films in the undoped state. FTIR results confirmed the presence of CdSt and undoped polyaniline (and 16-mer) in the transferred LB films, corroborating the UV-vis results. XRD has shown separated domains of CdSt and PANi (l6-mer also) in the LB films. Ellipsometry data indicated a thickness of 25 &#197 per deposited layer. The high uniformity in these mixed LB films was confirmed by optical microscopy and surface potential measurements. The electrical conductivity was approximately 10-4 to 10-5 S.cm-1 for both PANi and 16-mer. Mixed PANi/CdSt films were also exposed to X-ray irradiation. After a given dose rate there is a red shift in the UV-vis spectra from the 600 nm region to the 800 nm region, similar to the usual acid doping process observed in polyaniline. Humidity effects have a strong influence on the doping process. A comparison is made of the conductivity measurements made here with those reported in the literature.