This study aimed to evaluate the effect of supplementation of rumen-protected lysine (RPL) and methionine (RPM) during the close-up period on nutritional status of Holstein calves. Calves used in the present study were from multiparous cows which received either a control diet (CON, n = 7, metabolizable protein (MP): 1,292 g/d, metabolizable Lysine (mLys): 87.3 g/d, metabolizable Methionine (mMet): 28.8 g/d) or a treatment diet (TRT, n = 8, MP: 1,314 g/d, mLys: 102.5 g/d, mMet: 33.8 g/d) supplemented with RPL (AjiPro-L, Ajinomoto Animal Nutrition North America, Inc.) and RPM (Met Plus, Nisso Shoji Co.,Ltd.) from 21 d before the expected calving date. After birth, all calves received the same amount of colostrum and the same diet. Blood samples were collected from the jugular vein of the calves at 2 or 3, 7, 14, 28 and 63 d of age and blood biomarkers and plasma free amino acids were measured. Data were analyzed with Welch t-test in each sampling points. In TRT calves, total protein at 7 and 14 d of age, albumin at 2 or 3, 7 and 14 d of age, total cholesterol at 7 and 28 d of age and glucose at 2 or 3 d of age were significantly higher (P < 0.05) than CON calves. β-globulin, which is synthesized mainly in the liver and indicates the status of protein nutrition, tended to be higher (P < 0.10) at 2 or 3, 7 d of age and significantly higher (P < 0.05) at 14 and 28 d of age in TRT calves than CON calves. Total amino acids at 2 or 3 d of age was significantly higher (P < 0.05) in TRT calves than CON calves. These results suggest that the nutritional status was improved in TRT calves. Furthermore, γ-globulin at 7 d of age and γ-glutamyltransferase (GGT) at 7 and 14 d of age, respectively, was significantly higher (P < 0.05) in TRT calves than CON calves. That suggests TRT calves were in a better status of passive immunity. In conclusion, maternal supplementation of RPL and RPM during the close-up period may improve the nutritional status and passive immunity in Holstein calves.