This study examines the impact of moderate and high lighting and indoor air quality (IAQ) conditions on students’ well-being during a regular academic course in higher education. To determine the precise contribution of these two indoor environmental factors, students’ perceptions of their well-being were examined with the Positive and Negative Affect, Basic Emotional Process, and Karolinska Sleepiness Scale. Data were collected from 83 students, resulting in 285 responses, distributed across four combinations of moderate and high IAQ conditions, resp. > 800 ppm ≤ 950 ppm carbon dioxide (CO2) and < 800 ppm CO2, and moderate and high horizontal illuminance (HI) levels, resp. 500 lx and 750 lx. The results indicated that high HI levels did not enhance students’ perceived well-being compared to moderate levels. However, high IAQ conditions significantly contributes to students’ well-being, compared to moderate conditions. Interaction effects between the two factors were observed at moderate conditions.