The timber board interlayer is applied as the formwork for the pouring of concrete slab in various practical applications of timber–concrete composite structures, with the rehabilitation of timber buildings, in particular. At present, there are few studies performed to study the shear stiffness of inclined screws in timber–concrete composite beams with timber board interlayer. In this article, eight groups of shear tests were carried out to study the shear stiffness of inclined screws in timber–concrete composite beams with timber board interlayer. The key parameters included the embedment depth of the screw connector into timber, screw diameter, the thickness of concrete slab, and concrete strength. As indicated by the test results, the shear stiffness of the inclined screws was improved as the embedment depth of screw into timber and screw diameter increased. When the embedded depth of screw into concrete remained unchanged, the thickness of concrete slab and concrete strength exhibited no significant impact on the shear stiffness of inclined crossing screws. On the basis of the theory of a beam on a two-dimensional elastic foundation, the calculation method for predicting the shear stiffness of inclined screw in timber–concrete composite beams with interlayer was proposed. The comparisons demonstrated that the shear stiffness of inclined screw can be well predicted using the calculation method.