This study aims to determine the particulate matter (PM2.5) mass concentrations and the comfort parameters (total bacterial counts (TBC), total fungal counts (TFC), relative humidity and temperature) in a university building. The samplings were carried out in three different indoor areas, including lecture hall, laboratory and lecturer office. PM2.5 samples were collected over a period of 8 h sampling using a low volume sampler (LVS). The anemometer Model Kestrel 0855YEL was used to measure relative humidity and temperature parameters. The sampling of airborne microorganisms was conducted by using microbial sampler at 350 L air sampled volume. The results showed that the highest average of PM2.5 was at lecture hall (88.54 ± 26.21 µgm-3) followed by lecturer office (69.79 ± 19.06 µgm-3) and laboratory (47.92 ± 24.88 µgm-3). The mean of TBC and TFC readings recorded as follow; 32.71 ± 5.91 cfu m-3 and 76.71 ± 21.5 cfu m-3 for laboratory, 112.1 ± 29.06 cfu m-3 and 124.67 ± 23.35 cfu m-3 for lecturer office, 121.74 ± 19.33 cfu m-3 and 115.33 ± 8.08 cfu m-3 for lecture hall. The average of all comfort parameter was within the prescribed standard by Industry Code of Practice on Indoor Air Quality 2010 for all sampling sites. Therefore, all occupants of the building can work in a conducive and comfortable environment. This study is in line with the objectives of National Policy on the Environment (DASN), which focusing on achieving a clean, safe, healthy and productive environment for present and future generations.