This study was carried out to investigate the effect of DDGS hydrolysate (H-DDGS) and rumen-protected lysine-choline complex (RPLC) on milk production and blood metabolites in dairy cows. Feeding trials was performed to latin-square design using the 4 mid-lactational cows for 8 weeks, and treated with T1 (H-DDGS 1.1 kg), T2 (H-DDGS 0.73 kg + RPLC 0.15 kg), T3 (H-DDGS 0.37 kg + RPLC 0.30 kg) and T4 (H-DDGS 0.37 kg + RPLC 0.15 kg) according to the content of protein source. Dry matter intake (DMI) of TMR and average weight showed no significant difference between treatments. The milk production of T1, T2 and T4 were significantly higher than T3 treatment (p < 0.05), and milk/DMI efficiency tend to increase in the T1. Milk components showed no significant difference between treatments, however, the milk protein of T2 increased to 0.15% than T1. Also blood metabolites showed no significant difference between treatments. But T-CHO level numerically represented a lower trend in the treatments of adding to RPLC compared with T1. This result suggests that the high level (1.1 kg) of H-DDGS is expected to improve the feed utilization without the negative impact on weight gain, feed intake and milk production as the lactation stage of dairy cows proceeds, and 0.15 kg of RPLC under the same feeding conditions of H-DDGS may be useful on fat metabolism.