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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Evaluation of capillary chromatographic supports for immobilized human purine nucleoside phosphorylase in frontal affinity chromatography studies

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Autor(es):
de Moraes, Marcela Cristina [1] ; Temporini, Caterina [2] ; Calleri, Enrica [2] ; Bruni, Giovanna [3] ; Ducati, Rodrigo Gay [4] ; Santos, Diogenes Santiago [4] ; Cardoso, Carmen Lucia [5] ; Cass, Quezia Bezerra [1] ; Massolini, Gabriella [2]
Número total de Autores: 9
Afiliação do(s) autor(es):
[1] Univ Fed Sao Carlos, Dept Quim, BR-13565905 Sao Paulo - Brazil
[2] Univ Pavia, Dept Drug Sci, I-27100 Pavia - Italy
[3] Univ Pavia, Dept Chem, I-27100 Pavia - Italy
[4] Pontificia Univ Catolica Rio Grande do Sul PUCRS, Ctr Pesquisas Biol Mol & Func, Inst Nacl Ciencia & Tecnol TB, Porto Alegre, RS - Brazil
[5] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, BR-14040901 Sao Paulo - Brazil
Número total de Afiliações: 5
Tipo de documento: Artigo Científico
Fonte: Journal of Chromatography A; v. 1338, p. 77-84, APR 18 2014.
Citações Web of Science: 9
Resumo

The aim of this work was to optimize the preparation of a capillary human purine nucleoside phosphorylase (HsPNP) immobilized enzyme reactor (IMER) for characterization and affinity screening studies of new inhibitors by frontal affinity chromatography coupled to mass spectrometry (FAC-MS). For this purpose two monolithic supports, a Chromolith Speed Rod (0.1 mm I.D. x 5 cm) and a methacrylate-based monolithic epoxy polymeric capillary column (0.25 mm I.D. x 5 cm) with epoxy reactive groups were considered and compared to an IMER previously developed using an open fused silica capillary. Each HsPNP-IMER was characterized in terms of catalytic activity using Inosine as standard substrate. Furthermore, they were also explored for affinity ranking experiments. K-d determination was carried out with the based fused silica HsPNP-IMER and the results are herein discussed. (C) 2014 Elsevier B.V. All rights reserved. (AU)

Processo FAPESP: 08/04051-0 - Desenvolvimento e aplicação de biorreatores capilares para a triagem de inibidores da Purina Nucleosídeo Fosforilase
Beneficiário:Marcela Cristina de Moraes
Modalidade de apoio: Bolsas no Brasil - Doutorado