Artículos de revistas
Effect of IDA and TREN chelating agents and buffer systems on the purification of human IgG with immobilized nickel affinity membranes
Registro en:
Journal Of Chromatography B-analytical Technologies In The Biomedical And Life Sciences. Elsevier Science Bv, v. 861, n. 1, n. 64, n. 73, 2008.
1570-0232
WOS:000252569600009
10.1016/j.jchromb.2007.11.018
Autor
Ribeiro, MB
Vijayalakshmi, M
Todorova-Balvay, D
Bueno, SMA
Institución
Resumen
The purification of IgG from human plasma was studied by comparing two affinity membranes complexed with Ni(II), prepared by coupling iminodiacetic acid (IDA) and Tris(2-aminoethyl)amine (TREN) to poly(ethylenevinyl alcohol), PEVA, hollow fiber membranes. The Ni(II)-TREN-PEVA hollow fiber membrane had lower capacity for human IgG than the complex Ni(II)-IDA-PEVA, but with similar selectivity. The IgG in peak fractions eluted from the Ni(II)-IDA-PEVA with a stepwise concentration gradient of Tris-HCl pH 7.0 (100-700 mM) reached a purity of 98% (based on IgG, IgM, IgA, albumin, and transferrin nephelometric analysis). Adsorption IgG data at different temperatures (4-37 degrees C) were analyzed using Langmuir model resulting in a calculated maximum capacity at 25 degrees C of 204.6 mg of IgG/g of dry membrane. Decrease in K-d with increasing temperature (1.7 x 10(-5) to 5.3 x 10(-6) M) indicated an increase in affinity with increased temperature. The positive value of enthalpy change (26.2 kJ/mol) indicated that the adsorption of IgG in affinity membrane is endothermic. Therefore, lower temperature induces adsorption as verified experimentally. (c) 2007 Elsevier B.V. All rights reserved. 861 1 64 73