RESUMO
A label-free surface-enhanced Raman scattering (SERS) was designed for sensitive detection of interleukin-6 (IL-6). The sensing element composed of anti-IL-6 antibodies adsorbed on the surface of spherical gold nanoparticles (AuNPs) as SERS-active surface. The principle of detection was probing antibody conformational changes using its intrinsic SERS fingerprint after binding to IL-6. Comparison of SERS spectra of antibody before and after binding to IL-6 showed that secondary structure of antibody does not change upon binding to IL-6. Vibrational information from disulfide bonds ν(SS) in antibody structure indicated some changes of geometry around SS bridges as a consequence of the immunocomplex formation. Transmission electron microscopy (TEM) and UV-Vis spectroscopy were used to confirm AuNPs conjugation with antibody as well as IL-6 binding to antibody on the surface of AuNPs. The SERS-based immunoassay showed a wide linear range (2.0-1000 pg mL-1) and a high sensitivity with a limit of detection (LOD) as low as 0.91 pg mL-1 (0.04 pM) without using any extrinsic Raman label. UV-Vis spectroscopy was employed as a conventional method for IL-6 detection based on observation of any change in the position of localized surface plasmon resonance (LSPR) band of AuNPs-antibody conjugates with LOD of 10 ng mL-1.