Abstract: The aim of the study was to investigate the effects of pH, temperature, extraction time and flour/water ratio, monitored the combined effects of the above parameters through the use of Response Surface Methodology (RSM) during sesame seeds (Sesamum indicum L.) protein extraction, analyzed the highest protein recovered from the extraction for it amino acid profile. All the four parameters have influence on the extraction process of sesame seeds protein when they were varied individually and combined. The maximum recovery of protein during the extraction process was 94% at 12, 45°C, 45 min and 6/100 g mL-1 (w/v) of pH, temperature, time and ratio of sesame flour to water, respectively. The recovery was described by the model relationship given as Recovery (R) (extraction) = 51.33 + 2.75A + 1.25B + 2.00C - 3.58D +8.02A2 + 2.27B2 + 4.15C2 - 0.48D2 + 4.50AB + 0.000AC-1.00AD-1.00BC -1.00BD -0.75CD. The model showed good fit, since the R2 indicated that 96.47 % of the variability within the range of values studied could be explained by the model. The analysis of amino acids revealed the essential amino acids present in the highest extract were methionine, lysine, isoleucine, leucine, threonine, histidine, phenylalanine and tryptophan as 3.6, 2.6, 3.4, 7.1, 3.6, 2.2, 4.6 and 2.1, respectively. The extraction process was successfully done as RSM provided a good method in extracting protein from sesame seeds and obtains the optimum extraction conditions with manageable experimental runs and thus saved time and resources.