Biomolecules play critical roles in biological activities and are closely related to various disease conditions. The reliable, selective and sensitive detection of biomolecules holds much promise for specific and rapid biosensing. In recent years, luminescent lanthanide probes have been widely used for monitoring the activity of biomolecules owing to their long luminescence lifetimes and line-like emission which allow time-resolved and ratiometric analyses. In this review article, we concentrate on recent advances in the detection of biomolecule activities based on lanthanide luminescent systems, including upconversion luminescent nanoparticles, lanthanide-metal organic frameworks, and lanthanide organic complexes. We also introduce the latest remarkable accomplishments of lanthanide probes in the design principles and sensing mechanisms, as well as the forthcoming challenges and perspectives for practical achievements.