Article ID Journal Published Year Pages File Type
1988633 Journal of Biochemical and Biophysical Methods 2006 19 Pages PDF
Abstract

Terpolymer bead particles (100–350 μm in diameter) were prepared by suspension radical polymerization from methacrylate esters [2,3-epoxypropyl methacrylate (GMA), 2-(2-hydroxyethoxy)ethyl methacrylate (DEGMA) and ethylene dimethacrylate (EDMA)] and subsequently derivatized affording iminodiacetic acid (IDA) chelating sorbents. The sorbents differed in pore volumes (0–0.7 cm3/g) and specific surface areas (0.03–9.8 m2/g) of their matrices as well as in the amounts of immobilized Ni2+–IDA complexes (0.03–1.58 mmol/g). The binding of imidazole was studied by frontal chromatography to evaluate the accessibility of Ni2+–IDA complexes. It was found that an increase in the bonded imidazole content with increasing immobilized Ni2+–IDA concentration was strongly dependent on the matrix morphology. A higher pore volume of the matrix significantly improved the utilizability of Ni2+–IDA complexes for imidazole binding. The performance of the sorbents based on two porous matrices with immobilized Ni2+–IDA concentration (0.1–1.58 mmol/g) differing in pore size distributions was compared in immobilized metal affinity chromatography (IMAC) of monoclonal mouse immunoglobulin IgG1 specific against human choriogonadotropic hormone (GTH-spec IgG1). The results have shown that sorbents based on matrix with large pores (up to 20 μm in diameter) exhibited high protein binding capacities. The GTH-spec IgG1 (Mw = 158,000) was eluted from all the sorbents in its native form as was confirmed by MALDI-TOF.

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