Albumin-binding surfaces: synthesis and characterization.
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.
McFarland
C D
CD
Cooperative Research Centre for Cardiac Technology, CSIRO Molecular Science, Sydney Laboratory, North Ryde, NSW, Australia.
Jenkins
M
M
Griesser
H J
HJ
Chatelier
R C
RC
Steele
J G
JG
Underwood
P A
PA
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
D000911:Antibodies, Monoclonal; D020099:Coated Materials, Biocompatible; D008567:Membranes, Artificial; D012709:Serum Albumin; D001360:Avidin; D001710:Biotin; D011138:Polytetrafluoroethylene; D003101:Collodion