This study explored the potential for using seed cake from hemp (Cannabis sativa L.) as a protein feed for dairy cows. The aim was to evaluate the effects of increasing the proportion of hempseed cake (HC) in the diet on milk production and milk composition. Forty Swedish Red dairy cows were involved in a 5-week dose-response feeding trial. The cows were allocated randomly to one of four experimental diets containing on average 494 g/kg of grass silage and 506 g/kg of concentrate on a dry matter (DM) basis. Diets containing 0 g (HC0), 143 g (HC14), 233 g (HC23) or 318 g (HC32) HC/kg DM were achieved by replacing an increasing proportion of compound pellets with cold-pressed HC. Increasing the proportion of HC resulted in dietary crude protein (CP) concentrations ranging from 126 for HC0 to 195 g CP/kg DM for HC32. Further effects on the composition of the diet with increasing proportions of HC were higher fat and NDF and lower starch concentrations. There were no linear or quadratic effects on DM intake, but increasing the proportion of HC in the diet resulted in linear increases in fat and NDF intake, as well as CP intake (P < 0.001), and a linear decrease in starch intake (P < 0.001). The proportion of HC had significant quadratic effects on the yields of milk, energy-corrected milk (ECM) and milk protein, fat and lactose. The curvilinear response of all yield parameters indicated maximum production from cows fed diet HC14. Increasing the proportion of HC resulted in linear decreases in both milk protein and milk fat concentration (P = 0.005 and P = 0.017, respectively), a linear increase in milk urea (P < 0.001), and a linear decrease in CP efficiency (milk protein/CP intake; P < 0.001). In conclusion, the HC14 diet, corresponding to a dietary CP concentration of 157 g/kg DM, resulted in the maximum yields of milk and ECM by dairy cows in this study.