Since the discovery of graphene, a large number of two-dimensional (2D) materials have been found and studied. The charge carriers of 2D materials are restrained in a 1 nm physical space, which results in high sensitivity of charge carriers to chemical or electrical doping. It brings a technical innovation into a biosensing field. No matter what sensing mechanism the biosensor process is based on, it includes the process of detecting object recognition and signal transformation. The target recognition is normally realized by nano-bioprobes at the sensing interfaces of the devices. After the recognition, 2D materials at the biosensing interface can realize signal output. Constructing bioprobes and 2D materials at an atomic level at the biosensing interface can modulate the physical and chemical activity precisely in the process of sensing, which improves the sensing performances of devices. Here, we review the recent progress of constructing the 2D biosensing interfaces. Especially, we discuss various biosensing mechanisms and different nano-bioprobes. We also suggest the further research direction of this field.