The invention provides a manufacturing method of a thin film transistor substrate including: sequentially forming a gate electrode, a gate insulating layer covering the gate electrode, an active material layer, and a photo-sensitive material layer on a first substrate; performing a photolithography process by using a half tone mask to form a protective layer which is above the gate electrode and has a first recess and a second recess; wet etching the active material layer by using the protective layer as a mask to form an active layer; removing a portion of the protective layer at bottoms of the first recess and the second recess to expose a first portion and a second portion of the active layer respectively; forming a first electrode connecting to the first portion; and forming a second electrode connecting to the second portion.