The design and development of novel electrochemiluminescence (ECL) systems with near-infrared (NIR) emission beyond 800 nm are promising for ECL evolution, especially for improving the throughput of the spectrum-resolved multiplexing ECL assay and biological imaging. Herein a biocompatible and environmentally friendly luminophore, that is, the methionine-stabilized Au nanoclusters (Met-Au NCs), are proposed to achieve efficient aqueous ECL of ∼835 nm with triethanolamine as coreactant. The Met-Au NCs not only demonstrate a 75 times enhanced ECL compared with the traditional Au NCs with bovine serum albumin as a capping agent but also can be employed as ECL tags to label proteins with a methionine linker and enable a highly sensitive NIR ECL bioassay. A sandwich-type NIR ECL immunosensor is constructed with the Met-Au NCs as tags and α-fetoprotein (AFP) as a model analyte and exhibits a wide linearity range from 3 fg·mL-1 to 0.1 ng·mL-1 with a limit of detection of 1 fg mL-1 (S/N = 3) as well as desired selectivity.