Effect of the matrix structure and concentration of polymer-immobilized Ni2+-iminodiacetic acid complexes on retention of IgG1.
Terpolymer bead particles (100-350 microm 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 microm 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.
Krácalíková
Katerina
K
Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Heyrovsky Sq. 2, 162 06 Prague 6, Czech Republic. kmeszaro@imc.cas.cz
Tishchenko
Galina
G
Bleha
Miroslav
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D002614:Chelating Agents / Q000737:chemistry*; D006063:Chorionic Gonadotropin / Q000276:immunology; D002846:Chromatography, Affinity; D004852:Epoxy Compounds / Q000737:chemistry*; D006801:Humans; D007093:Imidazoles / Q000737:chemistry; D007098:Imino Acids / Q000737:chemistry*; D007074:Immunoglobulin G / Q000737:chemistry; D008689:Methacrylates / Q000737:chemistry*; D051379:Mice; D009532:Nickel; D011108:Polymers / Q000737:chemistry*; D016062:Porosity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
C512139:2-(2-hydroxyethoxy)ethyl methacrylate; D000911:Antibodies, Monoclonal; D002614:Chelating Agents; D006063:Chorionic Gonadotropin; D004852:Epoxy Compounds; D007093:Imidazoles; D007098:Imino Acids; D007074:Immunoglobulin G; D008689:Methacrylates; D011108:Polymers; C004919:ethylene dimethacrylate; C029899:imidazole; D009532:Nickel; C007870:glycidyl methacrylate; C008109:iminodiacetic acid