Effect of IDA and TREN chelating agents and buffer systems on the purification of human IgG with immobilized nickel affinity membranes

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2008, Vol.861 (1), p.64-73
• Main Authors: Ribeiro, Mariana Borsoi, Vijayalakshmi, Mookambesvaran, Todorova-Balvay, Daniele, Bueno, Sonia Maria Alves
• Format: Article
• Language: English
• Subjects: Adsorption; Affinity membrane; Analysis; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Buffers; Chelating Agents - chemistry; Chromatography, Affinity - instrumentation; Chromatography, Affinity - methods; Ethylenediamines - chemistry; Fundamental and applied biological sciences. Psychology; General pharmacology; Human IgG; Humans; IMAC; Imino Acids - chemistry; Immunoglobulin G - isolation & purification; Medical sciences; Nickel - chemistry; Pharmacology. Drug treatments; Purification; Temperature
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2007.11.018

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 °C) were analyzed using Langmuir model resulting in a calculated maximum capacity at 25 °C of 204.6 mg of IgG/g of dry membrane. Decrease in K d with increasing temperature (1.7 × 10 −5 to 5.3 × 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.