Two-dimensional periodic relief grating as a versatile platform for selective immunosorbent assay and visualizing of antigens.

In this study, we fabricated a nanopillar array of silicon oxide, involving very-large-scale integration (VLSI) and reactive ion etching (RIE), as two-dimensional periodic relief gratings (2DPRGs) on Si surfaces. Antihuman ALB was successively oriented on the pillar surface of 2DPRG modified protein G as an optical detector that is specific for targeted antigen. The antibody modified 2DPRG alone produces insignificant structure change, but upon immunocapture of antigens, the antigen filling in the 2DPRG leads to a dramatic change of the pillar scale. Binding of the antibodies to the 2DPRG occurs in a way that still allows them to function and selectively bind antigen. The performance of the sensor was evaluated by capturing HRP-human ALB on the antibody-modified 2DPRG and measuring the effective refractive index (neff) resulting from the attachment of antigens. The neff values of the 2DPRG are found to relate with the pillar scale of the 2DPRG, generated by antigen coupling, resulting in color change from pure green to orange, observed by the naked eye along an incident angle of 10-20°. Moreover, we calculated the filling factors inside the 2DPRG with effective-medium theory to verify the pillar structure changes. This technique eliminates much of the surface modifications and the secondary immunochemical or enzyme-linked steps that are common in immunoassays. Such films have potential applications as optical biosensors.