Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/198803
Type: Artigo de periódico
Title: Evaluation Of Immobilized Metal-ion Affinity Chromatography (imac) As A Technique For Igg(1) Monoclonal Antibodies Purification: The Effect Of Chelating Ligand And Support.
Author: Bresolin, I T L
Borsoi-Ribeiro, M
Tamashiro, W M S C
Augusto, E F P
Vijayalakshmi, M A
Bueno, S M A
Abstract: 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.
Subject: Adsorption
Antibodies, Monoclonal
Aspartic Acid
Chelating Agents
Chromatography, Affinity
Ethylenediamines
Imino Acids
Immunoglobulin G
Ligands
Membranes, Artificial
Metals
Nickel
Nitrilotriacetic Acid
Surface Properties
Zinc
Rights: fechado
Identifier DOI: 10.1007/s12010-009-8734-5
Address: http://www.ncbi.nlm.nih.gov/pubmed/19701728
Date Issue: 2010
Appears in Collections:Artigos e Materiais de Revistas Científicas - Unicamp

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