Ultra-sensitive detection of IgE using biofunctionalized nanoparticle-enhanced SPR

This paper describes an ultra-sensitive surface-based detection method using nanoparticle-enhanced surface plasmon resonance (SPR) for the detection of immunoglobulin E (IgE) proteins, which could potentially be used for the diagnosis of allergic diseases. Two different probes, anti-IgE and IgE specific aptamers, which can specifically interact with IgE at different epitopes were first investigated for their specific interaction with IgE using SPR. Langmuir adsorption coefficient (K_ads) values were measured as 2.0(±0.22) × 10⁸ M^-1 and 2.2(±0.20) × 10⁸ M^-1 for IgE interactions with anti-IgE and IgE specific aptamers, respectively. The SPR detection limit of the simple adsorption of IgE onto either anti-IgE or IgE specific aptamers was found to be about 1 nM. In order to improve the SPR detection signal for IgE, two different approaches utilizing surface formed sandwich complexes with biofunctionalized gold nanoparticles (Au-Nps) were designed and their detection performance were compared; the complexes were created via the adsorption of IgE onto (i) surface immobilized anti-IgE followed by the adsorption of IgE specific aptamer coated gold nanoparticles and (ii) IgE specific aptamer surface with the subsequent adsorption of anti-IgE coated gold nanoparticles. Both detection schemes were able to directly measure IgE at femtomolar concentrations.