A sensitive diagnostic assay of rheumatoid arthritis using three-dimensional ZnO nanorod structure

Abstract

We synthesized a three-dimensional nanorod structure of zinc oxide (ZnO) using a simple sol-gel process and systematically investigated properties of the ZnO nanorods regarding protein adsorption and effect on fluorescence emission. As compared to conventional polystyrene plate that has been widely used for strong protein adsorption, the ZnO nanorods had a superior protein adsorption capacity and significantly amplified fluorescence emission, suggesting the ZnO nanorods are attractive for fluorescence-based biomolecular detection assays. When applied to diagnostic assay of rheumatoid arthritis (RA) using cyclic citrullinated peptide (CCP) probe with a RCGRS motif that reportedly has a strong affinity for ZnO, the ZnO nanorods gave apparently high positive signals for all the RA-positive standards and patient sera, whereas upon the detection using conventional polystyrene plate, all the detection signals were relatively negligible. Moreover, the streptavidin-mediated immobilization of well oriented CCP further enhanced sensitivity, even for a 5000-times diluted patient serum. A highly sensitive detection of a very small amount of RA autoantibodies is important because individuals at high risk of developing RA can be identified several years before the clinical onset. Consequently, the fluorescence-based sensitive assay of RA was successfully performed using the three-dimensional ZnO nanorods, owing to the fluorescence amplification and protein/peptide adsorption properties and dimensionality of ZnO nanorods that in turn increases probe accessibility to anti-CCP RA autoantibodies. Although RA was assayed here for proof-of-concept, the ZnO nanorods-based assay can be applied in general to sensitive detection of a wide variety of antibody or protein targets. (C) 2011 Elsevier B.V. All rights reserved.