We aimed to assess postprandial metabolic and appetite responses to a mixed-macronutrient lunch following prior addition of whey protein to a carbohydrate-rich breakfast. Ten healthy males (age: 24 ± 1 y; body mass index (BMI): 24.5 ± 0.7 kg/m2) completed three trials in a non-isocaloric, crossover design. A carbohydrate-rich breakfast (93 g carbohydrate; 1799 kJ) was consumed with (CHO+WP) or without (CHO) 20 g whey protein isolate (373 kJ), or breakfast was omitted (NB). At 180 minutes, participants consumed a mixed-macronutrient lunch meal. Venous blood was sampled at 15 minute intervals following each meal and every 30 minutes thereafter, while subjective appetite sensations were collected every 30 minutes throughout. Post-breakfast insulinaemia was greater after CHO+WP (time-averaged area under the curve (AUC0-180 min): 193.1 ± 26.3 pmol/L), compared to CHO (154.7 ± 18.5 pmol/L) and NB (46.1 ± 8.0 pmol/L; p <0.05), with no difference in post breakfast (0-180 min) glycaemia (CHO+WP, 3.8 ± 0.2 mmol/L; CHO, 4.2 ± 0.2 mmol/L; NB, 4.2 ± 0.1 mmol/L; p = 0.247). There were no post lunch (0-180 min) effects of condition on glycaemia (p = 0.492), insulinaemia (p = 0.338) or subjective appetite (p > 0.05). Adding whey protein to a carbohydrate-rich breakfast enhanced the acute postprandial insulin response, without influencing metabolic or appetite responses following a subsequent mixed-macronutrient meal.