Beginning with the synthesis of carbon dots, continuing with their structure and characteristics, and concluding with their applications, which range from nanobiomedicine to electrochemical sensors, this chapter delves into the most significant and up-to-date developments in the utilization of carbon nanodots as key-role positions in the design of electrochemical sensors. Furthermore, recent advancements in the fields of nanotechnology and materials science have made it possible to synthesize a wide variety of novel materials that possess the morphologies that are wanted as well as the physicochemical qualities that are unlike any ever seen before. There has been a significant amount of interest paid to carbon-based nanomaterials, including carbon nanodots and quantum dots, as well as other forms of developed nanomaterials. This is mostly owing to the fact that these nanomaterials have the potential to be utilized in sensor applications, particularly in the electrochemical sensing of chemical molecules. The applications of carbon nanodots in the field of electrochemistry have recently attracted a lot of interest due to the favorable characteristics that they possess. These characteristics include hydrophilicity, facile functionalization, exceptional biocompatibility, intense luminescence, favorable solubility, high chemical stability, and negligible toxicity. They are hopeful candidates for a wide range of applications in a variety of sectors, including solar cells, bioimaging, optoelectronic devices, nanobiomedicine, and electrochemical domains, among others. Because of these one-of-a-kind features, it is possible to build carbon-based nanomaterials that are flexible and capable of sensitively detecting biological molecules. Carbon nanodots are being utilized in the design and utilization of electrochemical sensors as sensing materials for the electrochemical detection of various representative biological and pharmaceutical compounds. These compounds include rifampicin, alpha-fetoprotein, metal cations, cytokeratin fragment antigen 21-1, neuron-specific enolase, carcinoembryonic antigen, and others. A wide range of methodologies, such as advanced microfluidics, portable sensor chips, device downsizing, and microarray technology, will be utilized by these devices in order to carry out point-of-care testing.