This article focuses biosensors based on the electrohydrodynamic jet printing processes, including the fabrication process, micro/nano‐material characteristics, and the sensing principle. Electrohydrodynamic jet printing provides a novel way to fabricate micro/nano‐structures for flexible/wearable biosensors with excellent characteristics of low detection limits, high sensitivities, and fast responses. The nonwoven nanofibrous mats have been used as support substrates, detection elements, and sensor electrodes to enhance the sampling, reaction, and detection in the biosensing processes. Large‐scale electrospinning techniques are discussed to efficiently produce nanofibers for the industrial nanofiber applications. The near‐field electrospinning technique is also introduced which can overcome the bending instability in the nanofiber deposition process to make orderly nanofibrous patterns as the sacrificial structure for the fabrication of microfluidic chips as bioreactor chambers. The functional nanofibers increase the biosensing interfaces and sensing sensitivity with good flexibility to realize applications such as the vital signs detection for the fields of disease diagnosing, infection control, and health care. With these great advantages, electrohydrodynamic jet printing provides new opportunities for the development of point of care medical.