title: Albumin-binding surfaces: synthesis and characterization.
abstract: The nature of the proteinaceous film deposited on a biomaterial surface following implantation is a key determinant of the subsequent biological response. To achieve selectivity in the formation of this film, monoclonal antibodies have been coupled to a range of solid substrates using avidin-biotin technology. Antibody clones varied in their antigen-binding activity following insertion of biotin groups into lysine residues. Biotinylated antibodies coupled to solid substrates via an immobilized avidin bridge retained their biological activity. During immobilization of avidin a significant proportion of the protein molecules were passively adsorbed rather than covalently attached to the surface. This loosely bound material could be removed by stringent elution procedures which resulted in a surface density of 5.4 pmol avidin cm(-2). Although these conditions would be harsh enough to denature monoclonal antibodies, they did not destroy the biotin-binding activity of the residual surface-coupled avidin, enabling the subsequent immobilization of biotinylated antibodies. The two-step immobilization technique allowed the use of gentle protein modification procedures, reduced the risk of surface-induced denaturation and removed loosely bound material from the surface. The versatility of the technique encourages its application to a wide range of immobilization systems where retention of biological activity is a key requirement.
authors.affiliation: Cooperative Research Centre for Cardiac Technology, CSIRO Molecular Science, Sydney Laboratory, North Ryde, NSW, Australia.
authors.forename: C D
authors.initials: CD
authors.lastname: McFarland
authors.forename: M
authors.initials: M
authors.lastname: Jenkins
authors.forename: H J
authors.initials: HJ
authors.lastname: Griesser
authors.forename: R C
authors.initials: RC
authors.lastname: Chatelier
authors.forename: J G
authors.initials: JG
authors.lastname: Steele
authors.forename: P A
authors.initials: PA
authors.lastname: Underwood
mesh_terms: D000327:Adsorption; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism*; D001360:Avidin / Q000737:chemistry / Q000378:metabolism; D001710:Biotin / Q000737:chemistry; D020099:Coated Materials, Biocompatible / Q000138:chemical synthesis* / Q000378:metabolism*; D003101:Collodion; D005260:Female; D006801:Humans; D008567:Membranes, Artificial; D051379:Mice; D008807:Mice, Inbred BALB C; D011138:Polytetrafluoroethylene / Q000031:analogs & derivatives; D012709:Serum Albumin / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D013499:Surface Properties
chemical_list: D000911:Antibodies, Monoclonal; D020099:Coated Materials, Biocompatible; D008567:Membranes, Artificial; D012709:Serum Albumin; D001360:Avidin; D001710:Biotin; D011138:Polytetrafluoroethylene; D003101:Collodion