Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/75635
Type: Artigo de periódico
Title: Separation of human Fab fragments on negative mode Ni(II)-TREN-agarose chromatography
Author: da Silva, LCA
Serracchiani, MM
Miranda, EA
Bueno, SMA
Abstract: We evaluated the feasibility of using immobilized metal-ion affinity chromatography (IMAC) with nickel ion complexed with Tris(2-aminoethyl)amine (TREN) immobilized on agarose gel for purification of human Fab fragments by negative chromatography. Efficient purification of Fab fragments from digested human IgG (immunoglobulin G) (106.4% purity) was accomplished in Tris-HCI buffer at pH 7.5 without NaCI (based on total protein concentration and radial immunodiffusion of human Fab). A teChnological application of Ni(II)-TREN-agarose using non transgenic soybean protein extract spiked with human Fab fragments as feedstream was also studied. Experiments using Tris-HCI at pH 7.0 as loading buffer allowed the adsorption of almost all of the soybean proteins. Sixty-six percent of the loaded human Fab fragments were recovered in the flowthrough and washing fractions with about 90% purity. These results demonstrate that Ni(II)-TREN-agarose is a potential adsorbent for recombinant Fab fragment purification. (C) 2014 Published by Elsevier Ltd.
Subject: Human Fab fragments
Soybean proteins
IMAC
Purification
Ni(II)-TREN
Country: Inglaterra
Editor: Elsevier Sci Ltd
Rights: fechado
Identifier DOI: 10.1016/j.procbio.2013.12.006
Date Issue: 2014
Appears in Collections:Artigos e Materiais de Revistas Científicas - Unicamp

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