DNA-immobilized polyhydroxyethylmethacrylate microbeads for affinity sorption of human immunoglobulin G and anti-DNA antibodies.
Polyhydroxymethacrylate (PHEMA) microbeads were prepared by a suspension polymerization technique and activated by CNBr in an alkaline medium (pH 11.5). DNA molecules were immobilized onto CNBr-activated PHEMA beads. The amount of immobilized DNA was controlled by changing the medium pH and the initial concentrations of CNBr and DNA. The maximum DNA immobilization was observed at pH 5.0. Non-specific adsorption on the plain PHEMA microbeads was less than 0.1 mg/g. Much higher values, up to 2.75 mg/g, were achieved with the CNBr-activated PHEMA microbeads. Human immunoglobulin G (HIgG) adsorption onto PHEMA microbeads containing different amounts of DNA on their surfaces from aqueous solutions containing different amounts of HIgG at different pH values was investigated. The maximum HIgG adsorption was observed at pH 7.0. Non-specific HIgG adsorption onto the plain PHEMA microbeads was low (about 0.167 mg/g). Higher adsorption values, up to 7.5 mg/g, were obtained with the DNA-PHEMA beads. HIgG and anti-DNA antibody removal from the blood plasma obtained from a healthy donor and a patient with systemic lupus erythematosus (SLE) were also investigated. The maximum amounts of HIgG adsorbed from aqueous solution and human plasma onto the DNA-PHEMA microbeads were 7.35 and 23.46 mg/g, respectively. Anti-DNA antibody adsorption value was 40 mg/g.
Denizli
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Chemical Engineering Department, Hacettepe University, Ankara, Turkey.
Pişkin
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D000818:Animals; D000974:Antibodies, Antinuclear / Q000097:blood* / Q000737:chemistry; D002846:Chromatography, Affinity / Q000379:methods*; D003488:Cyanogen Bromide; D004247:DNA / Q000276:immunology*; D005399:Fishes; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000097:blood* / Q000737:chemistry; D008863:Microspheres; D011102:Polyhydroxyethyl Methacrylate / Q000737:chemistry*; D012996:Solutions
D000974:Antibodies, Antinuclear; D007074:Immunoglobulin G; D012996:Solutions; D011102:Polyhydroxyethyl Methacrylate; D004247:DNA; D003488:Cyanogen Bromide