FAO estimates that by 2050 food production must increase by 70% and meat production by 100% to address global demand. To meet this demand without exacerbating the environmental and public health costs of animal agriculture, growth in sustainable and alternative animal food products is critical. The overall goal of this project is to develop research-scaled novel protein concentrates to enhance value-added product marketing opportunities. The objective of this research was to compare the protein extracted with two methods, including alkaline and NaOH-Ethanol-Reducing agent methods regarding the extraction yield, and amino acid profiling to select the method with the higher extract yield and then the characterization of protein extracted with the selected method. The alkaline (NaOH) method showed an extraction yield of 1%, which was about 7.2% of the total protein content. Meanwhile, the protein content of the protein isolate was 56.11%, while the NaOH-Ethanol-Reducing Agent method revealed an extraction yield of 7.55%, which was 58.1% of the total protein. The protein content of sorghum protein isolate in this method was 88.83% which was significantly different from the NaOH method. Furthermore, the results from the amino acid composition analysis of sorghum showed that glutamic acid (2.82% w/w), leucine (1.82% w/w), and alanine (1.22% w/w) were the predominant amino acids, while protein extracted by NaOH from same sorghum, revealed that glutamic acid (7.44% w/w), arginine (5.06% w/w), and leucine (4.16% w/w) were the main amino acids, respectively. The amino acid composition analysis also revealed that despite the NaOH method, the protein extracted by the NER method did not show amino acids Taurine and Hydroxyproline. Moreover, glutamic acid, leucine, and alanine indicated the highest amount of 22.42%, 14.65%, and 9.43%, respectively, among other amino acids. The NER method was finally selected for functional properties evaluation as it showed a higher extraction yield and protein purity. The protein produced by NER method was further analyzed for its functional properties. The solubility of sorghum protein was evaluated at different pHs from 2 to 11, in which the highest protein solubility was at pH 8 to 11, while pH 2 to 6 revealed the lower solubility. Additionally, the water holding capacity (WHC) and oil holding capacity (OHC) of NER-generated protein also were evaluated, and the WHC was around 178% (1.78), which was lower than soybean and mung beans with 3.33 and 3.00, respectively, while the oil holding capacity of sorghum protein was 424%% (4.44), which was significantly higher than soybean and mung beans with 3.45 and 3.00, respectively.