The increasing potential of wearable electronics has been motivating the development of self-powered devices to overcome the restriction imposed by conventional power supply. Herein, a kind of highly electronegative and conducting material of MXene nanosheet has been innovatively integrated with poly(vinyl alcohol) (PVA) for electrospinning nanofibers film to fabricate flexible all-electrospun triboelectric nanogenerator (TENG). Silk fibroin (SF) was chosen for electrospinning nanofibers film to serve as electron donor for TENG with great triboelectricity and biocompatible nature. The MXene-based all-electrospun nanogenerator shows excellent stability and durability as well as extraordinary electrical performance, which supports an instantaneously maximum peak power density of 1087.6 mW/m2 as the load resistance is 5.0 MΩ. Moreover, beyond harvesting electrical power, the fabricated TENG was utilized for real-time monitoring various types of body motion. Lastly, we used the developed TENG to power electrowetting on dielectric (EWOD) chip to actuate droplet transport. Considering the excellent triboelectric performance, ease of large-scale manufacturing, and environmental friendliness, this MXene-based all-electrospun TENG provides a promising solution for developing practical, flexible, and self-powered electronic devices.