Antibody immobilization onto glow discharge treated polymers.
Previous studies have shown that certain glow discharge treated polymers strongly retain adsorbed albumin and fibrinogen. On the basis of this phenomenon, we have investigated the possibility of immobilizing antibodies on glow discharge treated surfaces for diagnostic immunoassay applications. As a model for antibody immobilization, bovine IgG was immobilized on the following polymers: polyethylene (PE), tetrafluoroethylene glow discharge treated PE (TFE/PE), poly(ethylene terephthalate) (PET), TFE/PET, poly(tetrafluoroethylene) (PTFE), ethylene glow discharge treated PET (E/PET) and hexamethyldisiloxane glow discharge treated PET (HMDS/PET). IgG was radiolabeled with 125I and immobilized by either of the following two methods: (a) physical adsorption of IgG on untreated and glow discharge treated polymers or (b) physical adsorption of albumin followed by chemical coupling of IgG to albumin by glutaraldehyde. IgG concentration as well as adsorption times were varied in order both to optimize the immobilization conditions and to investigate the adsorption and retention mechanisms. To evaluate the efficiency of the immobilization techniques, blood plasma, Tween-20, and sodium dodecyl sulfate (SDS) were used to elute the adsorbed IgG layer. We found that IgG was successfully immobilized on the fluorocarbon glow discharge treated surfaces by using either the physical adsorption or the glutaraldehyde coupling method, although the former is more efficient than the latter method.
Safranj
A
A
Center for Bioengineering, University of Washington, Seattle 98195.
Kiaei
D
D
Hoffman
A S
AS
D000418:Albumins / Q000737:chemistry; D000906:Antibodies / Q000737:chemistry*; D005976:Glutaral / Q000737:chemistry; D007074:Immunoglobulin G / Q000737:chemistry; D007164:Immunosorbents / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D013499:Surface Properties
D000418:Albumins; D000906:Antibodies; D007074:Immunoglobulin G; D007164:Immunosorbents; D011108:Polymers; D005976:Glutaral