Biotin is a water-soluble vitamin and serves as a coenzyme for carboxylases in human body. It is widely favored by chemists and biologists in recent years. In addition, it exhibits low toxicity in various physiological and pathological processes, and can be designed as a selectivity targeting drug carriers, which can efficiently deliver therapeutic drugs to cells. Nowadays, small molecules containing biotin have developed into a class of biofunctional molecules of high utility value. These compounds have the advantages of simple synthesis, easy functionalization, and high specificity. In this paper, the progress of biotin and its derivatives in biosensors, drug release and other fields are systematically reviewed, and the prospects for their development are presented.