Behavior of human immunoglobulin G adsorption onto immobilized Cu(II) affinity hollow-fiber membranes

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Iminodiacetic acid (IDA) and tris(2-aminoethyl)amine (TREN) chelating ligands were immobilized on poly(ethylene vinyl alcohol) (PEVA) hollow-fiber membranes after activation with epichlorohydrin or butanediol diglycidyl ether (bisoxirane). The affinity membranes complexed with Cu(II) were evaluated for adsorption of human immunoglobulin G (IgG). The effects of matrix activation and buffer system on adsorption of IgG were studied. Isotherms of batch IgG adsorption onto finely cut membranes showed that neither of the chelates, IDA-Cu(II) or TREN-Cu(II), had a Langmuirean behavior with negative cooperativity for IgG binding. A comparison of equilibrium and dynamic maximum capacities showed that the dynamic capacity for a mini-cartridge in a cross-flow filtration mode (52.5 and 298.4mgg(-1) dry weight for PEVA-TREN-Cu(II) and PEVA-IDA-Cu(II), respectively) was somewhat higher than the equilibrium capacity (9.2 and 73.3mgg(-1) dry weight for PEVA-TREN-Cu(II) and PEVA-IDA-Cu(II), respectively). When mini-cartridges were used, the dynamic adsorption capacity of IDA-Cu(II) was the same for both mini-cartridge and agarose gel. Copyright (c) 2013 John Wiley & Sons, Ltd.2610514520Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

The Effect Of Nacl On The Adsorption Of Human Igg Onto Cm-asp-peva Hollow Fiber Membrane-immobilized Nickel And Cobalt Metal Ions

Over the past decade, immobilized metal-affinity adsorbents have attracted increasing interest for purification of natural and recombinant immunoglobulin G (IgG). In this work, nickel and cobalt metal ions complexed with CM-Asp (carboxymethylaspartate) immobilized on poly(ethylenevinyl alcohol) (PEVA) hollow fiber membranes were evaluated for purification of human IgG from serum. The buffer system and NaCl had important effects on human serum protein adsorption in both adsorbents. Efficient purification of IgG was accomplished in sodium phosphate buffer without NaCl at pH 7.0. Under this condition, the electrostatic interactions are important for adsorption. The Ni(II)-CM-Asp-PEVA had a protein adsorption capacity of 17.5 mg of IgG mL-1 fiber when human serum diluted was loaded in crossflow filtration mode and the eluted IgG had a purity of 82.6 % (based on total protein and IgG, IgM, HSA, and Trf nephelometric analysis). Fitting the experimental IgG adsorption data to the Langmuir and Langmuir-Freundlich models showed that Ni(II)-CM-Asp and Co(II)-CM-Asp had Langmuirean and non-Langmuirean behavior, respectively, with positive cooperativity for IgG-Co(II)-CM-Asp binding, probably due to multipoint interactions (n = 2.12 ± 0.31). Thus, these membranes can be considered as alternative adsorbents for the purification or depletion of IgG from human serum. © 2014 Springer Science+Business Media New York.2005/06/15677688Altintas, E.B., Tuzmen, N., Uzun, L., Denizli, A., Immobilized metal affinity adsorption for antibody depletion from human serum with monosize beads (2007) Ind. Eng. Chem. Res., 46, pp. 7802-7810. , 10.1021/ie061164cAquino, L.C.L., Sousa, H.R.T., Miranda, E.A., Vilela, L., Bueno, S.M.A., Evaluation of IDA-PEVA hollow fiber membrane metal ion affinity chromatography for purification of a histidine-tagged human proinsulin (2006) J. Chromatogr. 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Evaluation of Immobilized Metal-Ion Affinity Chromatography (IMAC) as a Technique for IgG(1) Monoclonal Antibodies Purification: The Effect of Chelating Ligand and Support

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Monoclonal antibodies (MAbs) have been used for therapies and some analytical procedures as highly purified molecules. Many techniques have been applied and studied, focusing on monoclonal antibodies purification. In this study, an immobilized metal affinity chromatography membrane was developed and evaluated for the purification of anti-TNP IgG(1) mouse MAbs from cell culture supernatant after precipitation with a 50% saturated ammonium sulfate solution. The chelating ligands iminodiacetic acid, carboxymethylated aspartic acid (CM-Asp), nitrilotriacetic acid, and tris (carboxymethyl) ethylenediamine in agarose gels with immobilized Ni(II) and Zn(II) ions were compared for the adsorption and desorption of MAbs. The most promising chelating ligand-CM-Asp-was then coupled to poly(ethylene vinyl alcohol) (PEVA) hollow fiber membranes. According to SDS-PAGE and ELISA analyses, a higher selectivity and a purification factor of 85.9 (fraction eluted at 500 mM Tris) were obtained for IgG(1) using PEVA-CM-Asp-Zn(II). The anti-TNP MAb could be eluted under mild pH conditions causing no loss of antigen binding capacity.160721482165Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Adsorption of human serum proteins onto TREN-agarose: Purification of human IgG by negative chromatography

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Tris(2-aminoethyl)amine (TREN) - a chelating agent used in IMAC - immobilized onto agarose gel was evaluated for the purification of IgG from human serum by negative chromatography. A one-step purification process allowed the recovery of 73.3% of the loaded IgG in the nonretained fractions with purity of 90-95% (based on total protein concentration and nephelometric analysis of albumin, transferrin, and immunoglobulins A, G, and M). The binding capacity was relatively high (66.63 mg of human serum protein/mL). These results suggest that this negative chromatography is a potential technique for purification of IgG from human serum. (C) 2008 Elsevier B.V. All rights reserved.877416711723Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Water performance assessment of ETICS and influence on thermal conductivity, in Portuguese

PTDC/ECI-EGC/30681/2017Os sistemas de isolamento térmico pelo exterior do tipo ETICS são uma solução cada vez mais frequente na reabilitação de fachadas de edifícios em Portugal, devido não só a imposições legais, cada vez mais exigentes no que diz respeito à eficiência energética dos edifícios, mas também pelas vantagens que apresentam face a outras soluções. Ainda assim, estes sistemas compósitos possuem algumas desvantagens que continuam a condicionar o seu bom funcionamento e um estudo mais abrangente e focado na durabilidade ao longo do tempo do ETICS continua a ser necessário, devendo procurar minimizar-se anomalias na superfície e otimizar os sistemas de revestimento, evitando que o desempenho térmico da camada isolante seja afetado. Neste artigo, apresentam-se os primeiros resultados de um estudo de análise do desempenho à água de 12 soluções diferentes de ETICS, através de absorção capilar e secagem, procurando avaliar-se a capacidade de proteção à água dos acabamentos das diferentes soluções e o possível impacto de deficiências de impermeabilização na condutibilidade térmica do isolante. Todas as soluções cumprem o requisito do ETAG 004 que refere uma absorção capilar após 1 h de ensaio inferior a 1 kg/m2. Os resultados de secagem estão em concordância com os obtidos para a absorção de água por capilaridade: soluções que absorvem maior quantidade de água e mais rapidamente, também acabam por secar mais rapidamente. Ainda assim, a condutibilidade térmica dos isolantes (λ) é afetada pelo seu teor de água (Ψ), registando-se um aumento de λ com o aumento de Ψ. No entanto, este aumento não é significativo para a maioria dos isolantes, com exceção da lã mineral, onde λ aumenta muito rapidamente com o aumento de Ψ.authorsversionpublishe

Sentido do trabalho no discurso dos trabalhadores de uma ONG em Belo Horizonte The meaning of the work in the discourse of workers in a NGO from Belo Horizonte

Este artigo busca explicitar o discurso enunciado por trabalhadores do Comupra - Conselho Comunitário Unidos pelo Ribeiro de Abreu, uma ONG de Belo Horizonte, sobre a relação sentido positivo/prazer e negativo/sofrimento do trabalho. Além de fator de adoecimento ou saúde, o trabalho é central na vida dos indivíduos e da sociedade e é um dos campos de estudo atualmente em expansão. Porém, no Terceiro Setor, ainda é pouco estudado. Entrevistas semiestruturadas foram o principal instrumento de coleta de informações. O tratamento delas foi feito utilizando-se a análise do discurso. Seis trabalhadores do Comupra foram entrevistados. As análises revelaram que o sentido positivo do trabalho tem predominância sobre o negativo. As vivências que promovem o prazer são mais significativas do que aquelas que causam o sofrimento, mostrando que a escolha por trabalhar no Terceiro Setor está associada a uma expectativa de transformação pessoal e da comunidade.<br>The article's purpose is to reveal the discourse enunciated by the workers of the Comupra - Community Counsel United pro Ribeiro de Abreu, a NGO from Belo Horizonte, about the positive (pleasure) or negative (suffering) meaning they attribute to their work. Besides being a factor of illness and health, the work is central in individual and social lives and it is one of the fields of study presently in expansion. Yet, in the NGOs from the Third Sector, it is still not much studied. Semi-structured interviews were the main knowledge collection tool. The obtained information was treated by means of discourse analysis. Six workers from the Comupra were interviewed. The analyses revealed that the positive meaning of the work surpass the negative meaning. The situations in which pleasure occurs are taken as more important than those provoking suffering, showing that the choice for working in the third sector is associated to an expectation of individual and community changes