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Abstract(s)
A descoberta da mauveína por Perkin em 1856 marcou a indústria de síntese de corantes. Desde então, as cianinas têm vindo a ser sintetizadas com diversos fins, devido às características especiais que estas apresentam. Estas características passam pela elevada eficiência de fluorescência e coeficiente de extinção molar; a larga escala de comprimento de onda (c.d.o.) de absorção máxima; a facilidade em realizar a sua síntese; e a sua estabilidade relativamente elevada. Com aplicações em áreas como a biomédica e a fotodinâmica (entre outras), notou-se que a aplicação de cianinas enquanto ligandos em cromatografia de afinidade é na actualidade um processo pouco explorado.
A presente dissertação de mestrado, intitulada Síntese de Cianinas para Aplicação como Ligandos em Cromatografia de Afinidade, pretende verificar as interacções estabelecidas entre as indocarbocianinas sintetizadas e proteínas padrão na técnica de cromatografia de afinidade. Numa primeira fase realizou-se a síntese de três indocarbocianinas, na qual se realiza a introdução de grupos no azoto do núcleo base do indole presente em ambos os terminais das cianinas. Posteriormente, após caracterização dos compostos, realizou-se a imobilização das indocarbocianinas numa matriz polimérica, a celulose, realizando-se o estudo da interacção, de proteínas padrão, por cromatografia de afinidade. A técnica utilizada pretende verificar as interacções estabelecidas entre as indocarbocianinas como ligandos catiónicos, e proteínas padrão, lisozima, albumina sérica bovina (BSA) e α-quimotripsina, em diferentes condições (iónicas, pH e hidrofóbicas), com o objectivo de separar as proteínas em diferentes condições de eluição.
Neste trabalho serão apresentados e discutidos os resultados obtidos na síntese das indocarbocianinas bem como o seu comportamento como ligandos em cromatografia de afinidade.
Perkin´s mauve discovery in 1856 changed the dyes synthesis industry. Since this discovery, cyanines have been synthesized with several purposes, due to their special characteristics. These characteristics are such as: their fluorescence efficiency and molar extinction coefficient; their large range of wavelength of maximum absorption; their easy synthesis; and their relatively elevated stability. Applied to fields such as biomedical and photodynamic (between others), the lack of cyanine’s exploration as ligands at the process of affinity chromatography was noticed. This master's dissertation entitled Synthesis of Cyanine for Application as Ligands in Affinity Chromatography, intends to verify the interactions established between the synthesized indocarbocyanines and standard proteins through the technique of affinity chromatography. At the first stage of the investigation we preceded to the synthesis of three indocarbocyanines, during which took place the introduction of groups in the nitrogen of the indole’s core base present in both ends of the cyanine. Later, after characterization of the compounds, indocarbocyanines were immobilizated into a polymer matrix, cellulose, carrying out the study of the standard proteins interaction, through affinity chromatography technique. This technique aims to verify the interactions established between the indocarbocyanines as cationic ligands, and standard proteins, lysozyme, bovine serum albumin (BSA) and α-chymotrypsin, in different conditions (ionic, hydrophobic and pH), attempting to separate the proteins in different elution conditions. This paper will present and discuss the results obtained from the synthesis of indocarbocyanines as well as their behavior as ligands in affinity chromatography.
Perkin´s mauve discovery in 1856 changed the dyes synthesis industry. Since this discovery, cyanines have been synthesized with several purposes, due to their special characteristics. These characteristics are such as: their fluorescence efficiency and molar extinction coefficient; their large range of wavelength of maximum absorption; their easy synthesis; and their relatively elevated stability. Applied to fields such as biomedical and photodynamic (between others), the lack of cyanine’s exploration as ligands at the process of affinity chromatography was noticed. This master's dissertation entitled Synthesis of Cyanine for Application as Ligands in Affinity Chromatography, intends to verify the interactions established between the synthesized indocarbocyanines and standard proteins through the technique of affinity chromatography. At the first stage of the investigation we preceded to the synthesis of three indocarbocyanines, during which took place the introduction of groups in the nitrogen of the indole’s core base present in both ends of the cyanine. Later, after characterization of the compounds, indocarbocyanines were immobilizated into a polymer matrix, cellulose, carrying out the study of the standard proteins interaction, through affinity chromatography technique. This technique aims to verify the interactions established between the indocarbocyanines as cationic ligands, and standard proteins, lysozyme, bovine serum albumin (BSA) and α-chymotrypsin, in different conditions (ionic, hydrophobic and pH), attempting to separate the proteins in different elution conditions. This paper will present and discuss the results obtained from the synthesis of indocarbocyanines as well as their behavior as ligands in affinity chromatography.
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Keywords
Cromatografia de afinidade Indocarbocianinas
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Universidade da Beira Interior