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Effect of the matrix structure and concentration of polymer-immobilized Ni2+-iminodiacetic acid complexes on retention of IgG1
Krácalíková K, Tishchenko G, Bleha M.
Jazyk angličtina Země Nizozemsko
- MeSH
- chelátory chemie MeSH
- choriogonadotropin imunologie MeSH
- chromatografie afinitní MeSH
- epoxidové sloučeniny chemie MeSH
- financování organizované MeSH
- geny pro imunoglobuliny MeSH
- imidazoly chemie MeSH
- iminokyseliny chemie MeSH
- lidé MeSH
- methakryláty chemie MeSH
- monoklonální protilátky chemie MeSH
- myši MeSH
- nikl MeSH
- polymery chemie MeSH
- poréznost MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
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.
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- $a Kráčalíková, Kateřina. $7 _AN034157
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- $a Effect of the matrix structure and concentration of polymer-immobilized Ni2+-iminodiacetic acid complexes on retention of IgG1 / $c Krácalíková K, Tishchenko G, Bleha M.
- 314 __
- $a Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Heyrovsky Sq. 2, 162 06 Prague 6, Czech Republic. kmeszaro@imc.cas.cz
- 520 9_
- $a 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.
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