Immune complement activation on polystyrene and silicon dioxide surfaces. Impact of reversible IgG adsorption.
We have studied aspects of the molecular background to immune complement activation on solid surfaces. Quartz crystal microbalance with dissipation monitoring (QCM-D) sensor surfaces were modified by means of spin coating with polystyrene (PS) or sputtering of silicon dioxide (SiO2). The IC activation on modified QCM-D surfaces was investigated by incubation in serum, followed by determinations of the amounts of bound C3 fragments (C3c) at the surface. Determinations of soluble C3a and soluble C5b-9 complex (sC5b-9) were made with enzyme immunoassay (EIA) method. We found that IC activation was high on PS surfaces, independent of the method used for measurements. On the SiO2 surfaces, IC activation was generally lower, but still detectable with anti-C3c as well as sC5b-9 and C3a determinations. Pre-coating the surfaces with a layer of IgG resulted in that IC activation became very high on PS surface, while the IC response remained low on SiO2 surfaces. The lower level of IC activation on the SiO2 surfaces was explained by a low surface concentration of IgG as measured with QCM-D. This was a result of the high reversibility of the IgG protein adsorption as well as absence of sufficient conformational changes of adsorbed IgG molecules. The QCM-D method was as sensitive as the C3a and sC5b-9 determinations to reveal surface associated IC-activation on these model surfaces. Additional advantages of the QCM-D method are the broad dynamic measurement window, i.e. the high precision and the ability to perform time resolved measurements and the ease of making different surface modifications.
Sellborn
Anders
A
Department of Cell and Molecular Biology/Interface Biophysics, Lundberg Laboratory, Göteborg University, Box 462, SE-405 30 Göteborg, Sweden. anders.sellborn@gmm.gu.se
Andersson
Marcus
M
Hedlund
Julia
J
Andersson
Jonas
J
Berglin
Mattias
M
Elwing
Hans
H
D000327:Adsorption; D003167:Complement Activation; D015926:Complement C3a / Q000378:metabolism; D015938:Complement Membrane Attack Complex / Q000378:metabolism; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000378:metabolism*; D066298:In Vitro Techniques; D011137:Polystyrenes; D011791:Quartz; D012822:Silicon Dioxide; D012995:Solubility; D013499:Surface Properties
D015938:Complement Membrane Attack Complex; D007074:Immunoglobulin G; D011137:Polystyrenes; D011791:Quartz; D012822:Silicon Dioxide; D015926:Complement C3a