Sheep scab, caused by infestation of sheep skin with the mite Psoroptes ovis, is highly contagious, causing intense pruritus and represents a major welfare and economic concern. Current disease control strategies rely upon chemotherapy, however, its future sustainability is questionable due to issues of chemical residues, eco-toxicity and an emergence of acaricide resistance. The vaccination, an alternative control strategy, demonstrates protective immunity in sheep. To identify potential vaccine candidate antigens for P. Ovis, we adopted a two-pronged approach: antigens selection based on their interaction with host signalling pathways and the host immune-response; and those shown to be either immunogenic or involved in mite feeding activity. This resulted in the use and ultimate validation, in immunisation and challenge trials, of a seven recombinant protein sub-unit cocktail vaccine. There was strong evidence (p<0.002) that the vaccine reduced both lesion size and mite numbers following challenge in repeated protection trials. The mean lesion size in the immunised group was significantly smaller compared with the control group from 1 wpi until the end of the experiment at 6 wpi. All vaccine antigens elicited serum IgG responses following immunisation and prior to infestation, whereas animals in the control group did not produce antigen-specific IgG during the pre-infestation period. In addition, vaccinated animals showed an amnestic response, with levels of antigen-specific IgG against muGST, Pso o 1 and Pso o 2 increasing following infestation with P. ovis. This recombinant subunit vaccine represents the greatest reduction in lesion size to date, providing real encouragement for future production of a commercially-viable means of immunoprophylaxis.