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

Ayhan H., Kesenci K., Peskin E.

JOURNAL OF BIOMATERIALS SCIENCE-POLYMER EDITION, vol.11, no.1, pp.13-25, 2000 (SCI-Expanded) identifier


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 mg ml(-1); HMDA concentration, 4 mg ml(-1); and glutaraldehyde concentration, 0.070 mu g ml(-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 I-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 mg ml(-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 HIgG g(-1) polymer for Microbeads I and II, respectively.