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Biosensing technology, which aims to measure and control the signals of biological substances, has recently been developed rapidly due to increasing concerns about health and the environment. Top-down technologies have been used mainly with a focus on reducing the size of biomaterials to the nano-level. However, bottom-up technologies such as self-assembly can provide more opportunities to molecular-level arrangements such as directionality and the shape of biomaterials. In particular, block copolymers (BCPs) and their self-assembly have been significantly explored as an effective means of bottom-up technologies to achieve recent advances in molecular-level fine control and imaging technology. BCPs have been widely used in various biosensing research fields because they can artificially control highly complex nano-scale structures in a directionally controlled manner, and future application research based on interactions with biomolecules according to the development and synthesis of new BCP structures is greatly anticipated. Here, we comprehensively discuss the basic principles of BCPs technology, the current status of their applications in biosensing technology, and their limitations and future prospects. Rather than discussing a specific field in depth, this study comprehensively covers the overall content of BCPs as a biosensing platform, and through this, we hope to increase researchers' understanding of adjacent research fields and provide research inspiration, thereby bringing about great advances in the relevant research fields.