Two-dimensional photonic crystals (2D-PCs) fabricated on a cyclo-olefin polymer (COP) film using a printable photonics technology based on nano-imprint lithography (NIL) were used for label-free biosensing of insulin under wet conditions. In general, 2D-PC-based biosensing involves a complicated dry-up procedure after biosensing reactions on the 2D-PCs to obtain a high sensitivity through the large difference in refractive index. Therefore, it can be difficult to achieve simple operation involving single-step analysis. Performance of the biosensing under wet conditions would simplify the operational procedure. For label-free biosensing of insulin under wet conditions, the Fresnel reflection intensity change was used instead of the wavelength shift, which is the commonly used sensing signal. By detecting changes in refractive index caused by specific interactions between the antigen and antibody as the Fresnel reflection intensity changes, physiologically important concentrations of insulin could be detected, even under wet conditions. These results suggest that low-cost printed 2D-PCs offer great potential for single-step label-free biosensing through the introduction of a sample solution.

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The development of high-sensitive, and cost-effective novel biosensors have been strongly desired for future medical diagnostics. To develop novel biosensor, the authors focused on the specific optical characteristics of photonic crystal. In this study, a label-free optical biosensor, polymer-based two-dimensional photonic crystal (2D-PhC) film fabricated using nanoimprint lithography (NIL), was developed for detection of C-reactive protein (CRP) in human serum. The nano-hole array constructed NIL-based 2D-PhC (hole diameter: 230 nm, distance: 230, depth: 200 nm) was fabricated on a cyclo-olefin polymer (COP) film (100 µm) using thermal NIL and required surface modifications to reduce nonspecific adsorption of target proteins. Antigen-antibody reactions on the NIL-based 2D-PhC caused changes to the surrounding refractive index, which was monitored as reflection spectrum changes in the visible region. By using surface modified 2D-PhC, the calculated detection limit for CRP was 12.24 pg/mL at an extremely short reaction time (5 min) without the need for additional labeling procedures and secondary antibody. Furthermore, using the dual-functional random copolymer, CRP could be detected in a pooled blood serum diluted 100× with dramatic reduction of nonspecific adsorption. From these results, the NIL-based 2D-PhC film has great potential for development of an on-site, high-sensitivity, cost-effective, label-free biosensor for medical diagnostics applications.

SUMMARYFor the future medical diagnostics, high-sensitive, rapid, and cost effective biosensors to detect the biomarkers have been desired. In this study, the polymer-based two-dimensional photonic crystal (2D-PC) was fabricated using nanoimprint lithography (NIL) for biosensing application. In addition, for biosensing application, label-free detection of fibrinogen which is a biomarker to diagnose the chronic obstructive pulmonary disease (COPD) could be achieved using antigen-antibody reaction high-sensitively (detection limit: pg/ml order) and rapidly. Using this polymer-based 2D-PC, optical biosensor can be developed cost effectively. Furthermore, by using polymer as a base material for fabrication of 2D-PC, label-free detection of antigen-antibody reaction can be performed in visible region.

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