Protein A immobilization and HIgG adsorption onto porous/nonporous and swellable HEMA-incorporated polyEGDMA microspheres.
Both non swellable and swellable poly(EGDMA/HEMA) microbeads were produced by suspension copolymerization. These microbeads were modified by immobilization of a spacer-arm (hexamethylene diamine (HMDA)) and protein A. The optimal values for modifications were as follows: sodium periodate concentration, 1.0 mgml(-1); HMDA concentration, 4 mgml(-1); and glutaraldehyde concentration, 0.070 microgml(-1). Adsorption of protein A onto the plain and periodate oxidized poly(EGDMA/HEMA) microbeads were very close to each other, and were 0.01-0.02 mg protein A on the 1-g Microbeads I and II, respectively. Protein A immobilization on poly(EGDMA/HEMA) microbeads were studied at different temperatures, times, and pHs using single protein solution containing different amounts of proteins. The optimal values for immobilization were as follows: the initial protein A concentration, 0.1 mgml(-1); temperature, 25 degrees C; pH, 9.5; and immobilization time, 120 min. Incorporation of protein A resulted in 1.420 and 1.825 mg protein A on the 1-g Microbeads I and II, respectively. HIgG adsorption capacity on the protein A-incorporated poly(EGDMA/HEMA) microbeads is 27 and 35 mg HIgGg(-1) polymer for Microbeads I and II, respectively.
Ayhan
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Hacettepe University, Chemical Engineering Department, Ankara, Turkey.
Kesenci
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Pişkin
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D000327:Adsorption; D003432:Cross-Linking Reagents / Q000378:metabolism; D003959:Diamines / Q000493:pharmacokinetics; D004337:Drug Carriers; D005976:Glutaral / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000097:blood / Q000378:metabolism*; D008863:Microspheres; D010504:Periodic Acid / Q000378:metabolism; D011109:Polymethacrylic Acids / Q000138:chemical synthesis / Q000493:pharmacokinetics; D013205:Staphylococcal Protein A / Q000276:immunology / Q000378:metabolism*; D013696:Temperature
D003432:Cross-Linking Reagents; D003959:Diamines; D004337:Drug Carriers; D007074:Immunoglobulin G; D011109:Polymethacrylic Acids; D013205:Staphylococcal Protein A; C107571:poly(ethyleneglycol dimethacrylate-2-hydroxyethylmethacrylate); D010504:Periodic Acid; C009288:metaperiodate; D005976:Glutaral; C007297:1,6-diaminohexane