Protein A immobilization and HIgG adsorption onto porous/nonporous and swellable HEMA-incorporated polyEGDMA microspheres

Abstract

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.

Biocompatibility investigation and urea removal from blood by urease‐immobilized HEMA incorporated poly(ethyleneglycol dimethacrylate) microbeads

“…The hydroxyl groups on the poly(EGDMA/HEMA) microbeads were chemically oxidized with a three‐step procedure that included oxidation of hydroxyl groups on microbead surfaces, spacer‐arm incorporation, and finally glutaraldehyde treatment 8–10. Surface‐modified poly(EGDMA/HEMA) microbeads were used in enzyme immobilization experiments thereafter.…”

Poly(vinylferrocene-co-2-hydroxyethyl methacrylate) mediator as immobilized enzyme membrane for the fabrication of amperometric glucose sensor

“…VFc is a monomer having a redox potential approximately 0.4 V (vs. Ag/AgCl), and polyHEMA is a hydrophilic but a water-insoluble polymer [17]. HMDA is used as a spacer arm of immobilizing biomolecules such as enzymes [18] and protein A [19].…”