Purification of the specific immunoglobulin G1 by immobilized metal ion affinity chromatography using nickel complexes of chelating porous and nonporous polymeric sorbents based on poly(methacrylic esters). Effect of polymer structure.
Ni2+ complexes of the chelating nonporous and porous bead sorbents based on methacrylic esters crosslinked with ethylene dimethacrylate were used in isolation of the horseradish peroxidase-specific immunoglobulin IgG1 from the crude mouse ascitic fluid by immobilized metal ion affinity chromatography (IMAC). Iminodiacetic and aspartic acids were attached to porous poly(glycidyl methacrylate) beads differing in size, morphology and chemical composition. Ethylenediaminetriacetic acid and quinolin-8-ol chelating groups were attached mainly to the surface hydroxyl groups in nonporous poly(diethylene glycol methacrylate) beads through spacers. The latter sorbents exhibited better kinetic characteristics than the former but a very low IgG1 sorption capacity. In a single-step IMAC procedure, the best efficiency in the specific IgG1 purification was obtained with porous sorbents (recovery 92%, purity 73%). Differences in IMAC separations are discussed from the point of view of morphology of polymer beads as well as of the type and concentration of chelating ligands.
Tishchenko
Galina
G
Department of Polymer Membranes, Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Prague. tiscenko@imc.cas.cz
Dybal
Jirí
J
Mészárosová
Katerina
K
Sedláková
Zdenka
Z
Bleha
Miroslav
M
D002614:Chelating Agents / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004952:Esters / Q000737:chemistry; D007074:Immunoglobulin G / Q000302:isolation & purification*; D008854:Microscopy, Electron; D009532:Nickel / Q000737:chemistry*; D011109:Polymethacrylic Acids / Q000737:chemistry*
D002614:Chelating Agents; D004952:Esters; D007074:Immunoglobulin G; D011109:Polymethacrylic Acids; C030613:polymethacrylic acid; D009532:Nickel