Microporous poly(2-hydroxyethylmethacrylate) (pHEMA) films carrying cibacron blue F3GA, Congo red, and ferritin were prepared and used for iron(III) removal from human plasma. pHEMA films were produced by a photopolymerization of 2-hydroxyethylmethacrylate in the presence of azobisisobutyronitrile. The reactive dye ligands cibacron blue F3GA, Congo red, and bioligand ferritin were then covalently attached to the pHEMA films. The maximum dye loadings were 1.07 and 0.80 mu mol/cm(2) for cibacron blue and Congo red, respectively. The maximum amount of ferritin attached was 1.04 x 10(-3) mu mol/cm(2). Characterizations of the films were achieved by contact-angle, water-uptake, and scanning electron microscopy studies as well as atomic force microscopy images. The aqueous water-uptake properties and contact angles lair underwater of the pHEMA films did not change after derivatization with cibacron blue F3GA, Congo red, and ferritin. These hydrophilic films (contact angle = 45.3 degrees), having a swelling ratio of 58% (w/w) and carrying cibacron blue F3GA, Congo red, and ferritin, were used in Fe(III) removal studies. The maximum amounts of Fe(III) removed from human plasma by cibacron blue F3GA-, Congo red-, and ferritin-attached pHEMA films were 3.80 mug/cm(2) for cibacron blue F3GA, 4.41 mug/cm(2) for Congo red, and 8.1 mug/cm(2) for ferritin-attached films. Fe(III) ions could be repeatedly adsorbed and desorbed with these affinity pHEMA films without a noticeable loss in their Fe(III) adsorption capacity.