Catalytic Gold Deposition for Ultrasensitive Optical Immunosensing of Prostate Specific Antigen

Abstract

A major challenge in the development of bioanalytical methods is to achieve a rapid and robust quantification of disease biomarkers present at very low concentration levels in complex biological samples. An immunoassay platform is presented herein for ultrasensitive and fast detection of the prostate-specific antigen (PSA), a well-recognized cancer biomarker. A sandwich type immunosensor has been developed employing a detection antibody labeled with inorganic nanoparticles acting as tags for further indirect quantification of the analyte. The required high sensitivity is then achieved through a controlled gold deposition on the nanoparticle surface, carried out after completing the recognition step of the immunoassay, thus effectively amplifying the size of the nanoparticles from nm to µm range. Due to such an amplification procedure, quantification of the biomolecule could be carried out directly on the immunoassay plates using confocal microscopy for measurement of the reflected light produced by gold-enlarged nanostructures. The high specificity of the immunoassay was demonstrated with the addition of a major abundant protein in serum (albumin) at much higher concentrations. An extremely low detection limit for PSA quantification (LOD of 1.1 fg·mL−1 PSA) has been achieved. Such excellent LOD is 2–3 orders of magnitude lower than the clinically relevant PSA levels present in biological samples (4–10 ng·mL−1) and even to monitor eventual recurrence after clinical treatment of a prostate tumor (0.1 ng·mL−1). In fact, the broad dynamic range obtained (4 orders of magnitude) would allow the PSA quantification of diverse samples at very different relevant levels.