The development of sensors capable of characterizing and quantifying biomolecules is a significant contribution to the field of biology. In the past few decades, sensors have been routinely used in various sectors, especially in the field of medical diagnosis. In the fields of therapeutics and medicine, sensors have been used to fulfill the potential roles of clinically important biomarkers. Sensors used for diagnosing biomarkers or biomolecules are called “biosensors.” For biosensing applications, two important biomolecules are, the probe and the analyte. The probe used to interact with the target is called the “analyte” in the given test sample. The biosensor has the transducer, and physical elements to display the biomolecular interactions. In this chapter, the fundamentals of biosensors and biomolecules are overviewed.