Signal enhancement in a protein chip array using a 3-D nanosurface

Abstract

A silica based 3-D nanosurface was developed to enhance the signal intensity of a protein chip by increasing the surface density and reducing the aggregation of captured proteins immobilized on the nanosurface. The 3-D nanosurface was composed of silica nanopillar bundles formed from a nanoporous alumina template using the sol–gel method. The signal intensity of a protein spot increased exponentially when the capture probe was immobilized on a nanosurface with higher roughness and the amount of protein immobilized on the surface was proportional to the roughness of the nanosurface. To further investigate this nanosurface effect, changes in the nanosurface roughness before and after protein immobilization were investigated by AFM. The surface roughness was shown to increase after protein immobilization when the nanosurface initially had a relatively low surface roughness (Rq: 30–40 nm); however, the surface roughness decreased after protein immobilization when it initially had a high roughness (Rq: 60–130 nm). These results imply that a high nanosurface roughness decreases the overall aggregation of proteins on the surface. These findings were also confirmed by comparing the level of protein aggregation on nanosurfaces with high roughness and low roughness using AFM.