Since the development of enzyme electrodes, the research area of glucose biosensing has seen outstanding progress and improvement. Numerous sensing platforms have been developed based on different immobilization techniques and improved electron transfer between the enzyme and electrode. Interestingly, these platforms have consistently used innovative nanostructures and nanocomposites. In recent years, layered double hydroxides (LDHs) have become key tools in the field of analytical chemistry owing to their outstanding features and benefits, such as facile synthesis, cost-effectiveness, substantial surface area, excellent catalytic performance, and biocompatibility. LDHs are often synthesized as nanomaterial composites or manufactured with specific three-dimensional structures. The purpose of this review is to illustrate the biosensing prospects of LDH-based glucose sensors and the need for improvement. First, various clinical and conventional approaches for glucose determination are discussed. The definitions, types, and various synthetic methodologies of LDHs are then explained. Subsequently, we discuss the various research studies regarding LDH-based electrochemical and optical assays, focusing on modified systems, improved electron transfers pathways (through developments in surface science), and different sensing designs based on nanomaterials. Finally, a summary of the current limitations and future challenges in glucose analysis is described, which may facilitate further development and applications.