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Type: Artigo de periódico
Title: An aqueous two-phase system as a strategy for serum albumin depletion
Author: da Silva, MAO
Arruda, MAZ
Abstract: An aqueous two-phase system, based on the cloud point phenomenon, was proposed for blood plasma albumin depletion prior to gel electrophoresis (SDS-PAGE) analysis. Albumin removal was evaluated using a mixture of Triton (R) X-114 and SDS. Variables affecting phase separation and the partition coefficient (K), such as pH (from 5.0 to 7.6)and salt composition, were first optimized to carry out phase separation at room temperature and to promote albumin removal (15 min with ca. 95% efficiency) to the surfactant-rich phase, in a global process that results in reduction of 12%alpha-helix structures, confirmed through circular dichroism analysis. Gel electrophoresis analyses were carried out to compare crude human plasma composition with human plasma submitted to the cloud point procedure as well as the same plasma submitted to the separation of albumin using a commercially available system based on a chromatographic affinity column. The cloud point extraction procedure allowed identification of 18 protein bands present in the surfactant-poor phase, resulting in a more detailed electrophoretic profile, when compared with crude plasma (10 protein bands). (C) 2008 Elsevier B.V. All rights reserved.
Subject: Albumin depletion
Cloud point
Gel electrophoresis
Circular dichroism
Country: Holanda
Editor: Elsevier Science Bv
Rights: fechado
Identifier DOI: 10.1016/j.talanta.2008.07.055
Date Issue: 2009
Appears in Collections:Unicamp - Artigos e Outros Documentos

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