title: Development of affinity microparticles for extracorporeal blood purification based on crystalline bacterial cell surface proteins.
abstract: In this article, the development of specific adsorbents for extracorporeal blood purification are described. Affinity microparticles were prepared by linking Protein A to crystalline cell surface layers (S-layers) from Thermoanaerobacter thermohydrosulfuricus 1111-69. S-layers were used in the form of cell wall fragments obtained by breaking whole cells by ultrasonification, resulting in cup-shaped structures (average size 0.5 x 1 microm) completely covered with S-layer protein. Protein A was covalently bound to carboxylic acid groups of the S-layer protein after activation with 1-ethyl-3,3'(dimethylamino)propylcarbodiimide. In batch adsorption experiments with fresh frozen human plasma, the resulting S-layer based affinity microparticles showed a high adsorption capacity for IgG (40 mg IgG were bound per g wet pellet of S-layer based affinity microparticles). Fractions eluted from the microparticles were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. They contained only IgG demonstrating that adsorption was specific. In biocompatibility tests, preparations of the S-layer microparticles showed no low-density lipoprotein-reactivity, no cytotoxicity, and no cytokine inducing activity.
authors.affiliation: Christian Doppler Laboratory for Specific Adsorption Technologies in Medicine, Centre for Biomedical Technology, Donau-University Krems, Austria. weber@donau-uni.ac.at
authors.forename: V
authors.initials: V
authors.lastname: Weber
authors.forename: S
authors.initials: S
authors.lastname: Weigert
authors.forename: M
authors.initials: M
authors.lastname: Sára
authors.forename: U B
authors.initials: UB
authors.lastname: Sleytr
authors.forename: D
authors.initials: D
authors.lastname: Falkenhagen
mesh_terms: D000327:Adsorption; D001426:Bacterial Proteins / Q000097:blood* / Q000493:pharmacokinetics*; D001781:Blood Component Removal / Q000379:methods*; D003460:Crystallization; D005112:Extracorporeal Circulation; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood / Q000737:chemistry; D008565:Membrane Proteins / Q000097:blood* / Q000493:pharmacokinetics*; D013205:Staphylococcal Protein A / Q000097:blood*
chemical_list: D001426:Bacterial Proteins; D007074:Immunoglobulin G; D008565:Membrane Proteins; D013205:Staphylococcal Protein A