Application of a thermophoretic immunoassay in the diagnosis of lupus using outer membrane particles from E. coli

Abstract

Thermophoresis is the physical diffusion of molecules from hot to cold induced by a thermal gradient. Thermophoresis has been used to evaluate the interaction of biomolecules in solution. In this study, the outer membrane from E. coli was isolated and used to produce OM particles with a diameter of approximately 100 nm. These prepared OM particles were applied in a thermophoretic immunoassay. First, outer membrane (OM) particles with lipopolysaccharides (LPS) and anti-LPS antibodies were used as a model to demonstrate proof of concept and the difference in E. coli thermophoresis was explained by the changes in the molecular surface area (A) and effective charge (sigma(eff)). The hydrodynamic size of the molecules was measured as a changing parameter, molecular surface area (A), by dynamic laser scattering (DLS), and the zeta potential was measured as a changing parameter of effective charge (sigma(eff) ) and then evaluated by the Soret equation. Using the hydrodynamic size and zeta potential values, the interaction between the antigen (OM particle with LPS) and antibody (anti-LPS antibodies) could be monitored and the results were fitted to the thermophoretic immunoassay using the Soret coefficient and equation. Finally, this OM-based immunoassay was applied to the medical diagnosis of systemic lupus erythematosus. Here, OM particles with Ro and La proteins were used to analyze the autoantibodies in patient and control sera. Thermophoretic immunoassay results were also compared to the fitted analysis using hydrodynamic size and zeta potential values and the Soret coefficient and equation.