title: Novel plasma-separation dilayer gellan-gellan-sulfate adsorber for direct removal of extra domain A containing fibronectin from the blood of rheumatoid arthritis patients.
abstract: Rheumatoid arthritis (RA) patients, in whom cryogelation occurs in the presence of heparin, exhibit abnormally high concentrations of extra domain A containing fibronectin [EDA(+)FN] in their plasma. The selective removal of EDA(+)FN from patient blood is therefore of potential therapeutic benefit. Gellan-sulfate is a candidate ligand for the removal of EDA(+)FN due to its high affinity for FN. In this study, we prepare a novel adsorber for the direct removal of EDA(+)FN from patient blood. The adsorber has both a plasma separation function and EDA(+)FN trapping zones, and is prepared by cross-linking gellan-sulfate with epichlorohydrine. The ratio of gellan-sulfate to gellan in the adsorber is 48%. The surface and internal structure of gellan beads were observed by a range of microscopic techniques, and the beads were found to have a dilayer structure, consisting of a porous outer layer and an underlying gellan-sulfate phase as the adsorber. The affinity constants of the gellan-sulfate beads for EDA(+)FN were almost the same in blood as in buffer because the porous gellan coating acts to separate plasma from the cellular fraction of the blood. The removal rate of plasma proteins and blood cells from mock RA blood was measured for coated and uncoated gellan-sulfate beads. Removal rates were 30-32% for EDA(+)FN, 6-10% for fibrinogen, 10-14% for antithrombin III, 8% for C3, 4-7% for C4, and 0% for albumin. The removal rates of uncoated beads were 11% for white blood cells, 0% for red blood cells and 33% for platelets, whereas removal rates of 0% for white blood cells, 0% for red blood cells and 20% for platelets were achieved for coated beads. The coating effectively inhibits the adsorption of white blood cells and platelets. Existing problems with direct adsorbers, including selectivity and plasma separation, have been solved by this material.
authors.affiliation: Department of Chemistry for Materials, Faculty of Engineering, Mie University, 1515 Kamihama-Chou, Tsu, Mie 514-8507, Japan. miyamoto@chem.mie-u.ac.jp
authors.forename: Keiichi
authors.initials: K
authors.lastname: Miyamoto
authors.forename: Katsuyuki
authors.initials: K
authors.lastname: Sugihara
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Abe
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Nobori
authors.forename: Masayuki
authors.initials: M
authors.lastname: Tokita
authors.forename: Takashi
authors.initials: T
authors.lastname: Komai
mesh_terms: D000327:Adsorption; D001172:Arthritis, Rheumatoid / Q000097:blood*; D001781:Blood Component Removal / Q000379:methods*; D005353:Fibronectins / Q000097:blood*; D006801:Humans; D011134:Polysaccharides / Q000737:chemistry; D011135:Polysaccharides, Bacterial / Q000737:chemistry; D017434:Protein Structure, Tertiary; D013463:Sulfuric Acid Esters / Q000737:chemistry
chemical_list: D005353:Fibronectins; D011134:Polysaccharides; D011135:Polysaccharides, Bacterial; D013463:Sulfuric Acid Esters; C470030:gellan sulfate; C048288:gellan gum