Microalgae are rich in valuable biomolecules and grow on non-arable land with rapid growth rate, which has a host of new possibility as alternative protein sources. In the present study, extraction of proteins from Chlorella vulgaris via an efficient technique, Liquid Triphasic Flotation (LTF) system, was studied. The optimized conditions in LTF system were 70% v/v of t-butanol, 40% w/v of salt solution, 0.5% w/v of biomass, pH 5.54, 1:1 of salt to t-butanol solution, and 10 min of air flotation time to attain 87.23% of protein recovery and 56.72% of separation efficiency. Besides, the study on recycling t-butanol has demonstrated that only one run was sufficient to maintain the performance of system. The efficiency of LTF in extracting protein has performed better than just Three Phase Partitioning (TPP) system. LTF system is hence an effective protein extraction and purification method with minimum operation unit and processing time.