One hundred and thirteen mid-lactation cows fed same diets and supplemented with 20 g/d rumen-protected methionine (RPM) for 8 weeks were used to investigate the individual responses of dairy cows to RPM in terms of lactation performance, amino acids (AA) metabolism, and milk metabolites. Among the cows, 10 cows exhibited positive responses (PR) and 10 cows showed limited responses (LR) in energy-corrected milk yield to RPM were used for further analysis. The lactation performance changed from gradual decline to steady increase in PR cows, while kept downward trend in LR cows following RPM supplementation. In PR cows, the AA metabolism was notably enhanced after RPM supplementation, evidenced by increased mammary blood flow (69.4%, P = 0.05), mammary uptake and clearance rate and uptake-to-output ratio (U:O) of essential AA. The improved AA metabolism could be attributed to the enrichment of pyrimidine (P = 0.06) and pyruvate (P = 0.07) metabolism pathways, which may have stimulated mammary cell proliferation and enhanced AA uptake. Additionally, the upregulation of milk biotin (fold change > 2, variable importance projection > 1), known to support milk yield, likely contributed to the PR observed in PR cows. Conversely, in LR cows, RPM supplementation did not improve AA metabolism, decrease was observed in mammary uptake, mammary clearance rate, and U:O of cysteine, potentially due to cysteine being irreversibly converted from methionine. Moreover, the enrichment of central carbon metabolism in cancer pathway (P = 0.06), which also utilize methionine, along with the lysine degradation pathway (P = 0.04), suggests that methionine in the mammary glands may have been diverted toward non-lactational metabolic processes, resulting in absence of PR in LR cows. Our results indicate that the responses to RPM in dairy cows are individualized, with variation in lactation performance likely driven by differential AA metabolism.