Adsorção de lactoferrina em coluna supermacroporosa de criogel de poliacrilamida ativada com íons de cobre

• Ano de defesa: 2010
• Autor(a) principal: Carvalho, Bruna Mara Aparecida de
• Orientador(a): Minim, Luis Antonio
• Banca de defesa: Silva, Maria do Carmo Hespanhol da , Saraiva, Sérgio Henriques
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de Viçosa
• Programa de Pós-Graduação: Doutorado em Ciência e Tecnologia de Alimentos
• Departamento: Ciência de Alimentos; Tecnologia de Alimentos; Engenharia de Alimentos
• País: BR
• Palavras-chave em Português: Lactoferrina; Cromatografia por afinidade; Monolitos; Criogel
• Palavras-chave em Inglês: Lactoferrin; Affinity chromatography; Monoliths; Criogel
• Área do conhecimento CNPq: CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::TECNOLOGIA DE ALIMENTOS
• Link de acesso: http://locus.ufv.br/handle/123456789/432

In this study we evaluated the direct capture of lactoferrin from cheese whey ultrafiltrate in columns of supermacroporosas criogel by immobilized metal on affinity chromatography (IMAC). We also studied the adsorption of bovine serum lactoferrin by IMAC varying conditions of salt concentration, pH and temperature. With the experimental data and microcalorimetric measurements obtained the thermodynamic analysis was conducted of the process. It also assessed the influence of isotherm models, Langmuir, Langmuir-Freundlich, Temkin, Freundlich, and SMA (steric-mass action), the simulation of the adsorption process and the optimization of the adsorption of lactoferrin in IMAC. The data obtained from analysis of total protein, SDS-PAGE and molecular exclusion chromatography showed the high affinity of interactionm between lactoferrin and the copper ions immobilized on the array. From the results obtained by studying the adsorption of lactoferrin in criogel supermacroporoso was possible to observe that this process is dependent on the salt concentration, pH and ionic strength. Adsorption being inversely proportional to the increase of salt concentration and directly proportional to the increase of pH and temperature. Concerning the influence of models, the studies indicated isotherm models of Langmuir- Freundlich and SMA better for the simulations. The adsorption of lactoferrin from cheese whey on an cryogel- IDA-Cu2+ adsorbent was investigated using isothermal titration microcalorimetry to measure thermodynamic information regarding such processes. Isotherms data were measured at temperatures of 20° and °C, pH 6,0 and 8,0 and ionic strengths 200 e 1000mM NaCl. The equilibrium data were fit using the Langmuir model and the process was observed to be reversible. The temperature and pH positively affected the interaction of protein and adsorbent. Microcalorimetric studies indicated endothermic enthalpy of adsorption for some cases (20 ° C and 40 ° C pH 8.0, 1000mm) and the adsorption process entropically driven to such conditions.