title: Immobilisation of IgG onto gold surfaces and its interaction with anti-IgG studied by surface plasmon resonance.
abstract: The optical excitation of surface plasmon resonance (SPR) at a metal dielectric interface has been used to study the binding of immunoglobulin G (IgG) to gold and anti-IgG to immobilised IgG layers. In these studies both a monoclonal mouse and polyclonal sheep IgG were used as receptor layers for anti-IgG. The kinetics of binding were investigated by monitoring the reflectivity of light at an angle close to plasmon resonance. Both the initial rate of change and final reflectivity were measured during and after protein binding. The amount of protein bound to the surface was found to be less for the monoclonal mouse IgG compared to the polyclonal sheep IgG, these two IgG nominally being of the same dimensions and molecular weight. Further, anti-IgG binding produced greater changes in reflectivity than the initial IgG layers. By fitting the full angle-dependent reflectivity data to the Fresnel equation the effective protein layer thicknesses of IgG and anti-IgG as a function of concentration were determined. Differences in the effective thickness of the bound layer for the two IgG was observed, the mouse IgG having a thinner effective thickness compared with the sheep IgG. The limitations of direct binding of protein to metal surfaces in SPR biosensor applications are discussed.
authors.affiliation: CSIRO, Division of Chemical and Polymers, Clayton, Vic., Australia.
authors.forename: N J
authors.initials: NJ
authors.lastname: Geddes
authors.forename: A S
authors.initials: AS
authors.lastname: Martin
authors.forename: F
authors.initials: F
authors.lastname: Caruso
authors.forename: R S
authors.initials: RS
authors.lastname: Urquhart
authors.forename: D N
authors.initials: DN
authors.lastname: Furlong
authors.forename: J R
authors.initials: JR
authors.lastname: Sambles
authors.forename: K A
authors.initials: KA
authors.lastname: Than
authors.forename: J A
authors.initials: JA
authors.lastname: Edgar
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015374:Biosensing Techniques; D006046:Gold / Q000276:immunology*; D007074:Immunoglobulin G / Q000276:immunology*; D051379:Mice; D055095:Optics and Photonics; D012756:Sheep
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D006046:Gold