We develop highly sensitive graphene-based surface-enhanced Raman spectroscopy (SERS) substrates where monolayer graphene is coated on the Ag/mica substrates. SERS has attracted attention in biosensor applications that allow label-free detection with high specificity. Recently two-dimensional (2D) materials such as graphene, h-BN, and MoS2 have been explored as a new platform for SERS substrates. These materials have the advantage of being non-metal, with excellent biological compatibility, endurance, and uniformity characteristics. Then, we study the stacked SERS substrates where monolayer graphene is transferred on the mica substrates with single-crystal Ag thin films deposited on the back to improve SERS efficiency. The fabricated SERS substrates show stronger Raman signals than the mica substrates with Ag thin films. Then sandwich enzyme-linked immunosorbent assay (ELISA) is fabricated on the SERS substrates to detect bovine interleukin-6 (IL-6), inciting bovine mastitis.
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