The invention discloses a manufacturing method of a flexible OLED substrate. The manufacturing method comprises the following steps of S1, providing a supporting plate; S2, forming a flexible materiallayer on the supporting plate; S3, forming a barrier layer on the flexible material layer, wherein the barrier layer adopts a multi-layered structure, and comprises an inorganic layer and an organiclayer stacked mutually, wherein the inorganic layer is arranged on the surface on one side, far from the flexible material layer, of the barrier layer; the inorganic layer adopts the material of SiN<x> or SiO<x>N<y> with thickness of greater than 2,000-angstrom; and S4, manufacturing a layer of inorganic material to form a flat layer on the barrier layer through a bias sputtering method, removingconcave and convex points from the surface of the whole flexible material layer to realize an isoplanar structure. According to the manufacturing method, after the barrier layer is completely formed,the inorganic flat layer is manufactured by the bias sputtering method; and the tiny concave and convex points on the surface of the flexible material layer can be eliminated through etching and re-sputtering effects, so that electrode wiring smooth transition can be facilitated, post processing can be reduced, and the electrode wire breaking probability in device bending can be lowered.