A strategy for clean fuel by selective adsorption processing was deemed to be convenient and environmental-friendly in past decades. However, the development of adsorption desulfurization was tremendously subject to the fabrication of high-performance adsorbents with large capacity and high stability. Herein, we designed a novel route to fabricate the cloth-like carbon nanofiber film with a hierarchical porous structure by electrospinning. The structure and properties of the cloth-like carbon nanofiber films were determined by a series of characterizations. Subsequently, the desulfurization performance of the cloth-like carbon nanofiber films was examined by the simulated thiophene (TH) oil. Furthermore, the effect of adsorption conditions on the adsorption capacity was intensively investigated, such as carbonization temperature, initial concentration and desulfurization temperature. The results found that at optimal calcination temperature of 700 °C, the cloth-like carbon nanofiber films possessed the highest micropore volume (Vmic = 0.185 m3/g) and adsorption capacity (qe = 96.6 mg/g) at 800 mg/L initial concentration under the adsorption temperature of 25 °C. The results corroborated that the physical properties of the cloth-like carbon nanofiber films with the surface area of 417.8 m2/g, the total pore volume of 0.187 cm3/g and average pore diameter of 1.36 nm had an important influence on the high adsorption capacity. On this basis, the adsorption experimental data were best fitted to pseudo-second-order kinetic and Langmuir isotherm models. Furthermore, the other highlight of the cloth-like carbon nanofiber films was convenient for the separation from oil, thus achieving the desirable reused performance.