Cervical cancer was the fourth-highest incidence of female cancer globally in 2020 and the second-highest incidence of female cancer in Indonesia in 2019. Chemotherapy frequently causes side effects and resistance; thus, cancer therapeutic agents must be developed to find selective anticancer drugs. One of the plants that has the potential as an anticancer agent is soursop. Soursop seed extract demonstrated anticancer activity, but research on soursop seed protein has never been published. This research aims to determine the cytotoxicity of soursop seed protein (Annona muricata L.) isolated using DEAE matrix against HeLa and Vero cells. Soursop seeds were extracted by adding 0.14 M NaCl at 4 °C in 5 mM sodium phosphate buffer pH 7.2. Protein purification was done by ion-exchange column chromatography with stationary phase DEAE matrix and mobile phase NaCl ranging from the lowest molarity of 0.2 M to the highest molarity of 0.4 M. Protein fraction concentrations were measured using Biodrop at A260/280. MTT assay was employed to test the cytotoxic activity of protein fraction soursop seed, followed by measurement of cells' metabolic activity with ELISA reader at λ 550 nm. The results of protein isolation showed that the highest concentration of protein fraction eluted 0.2 M NaCl were 8466.2 μg/mL; 8346.4 μg/mL; 8205.0 μg/mL; and 8284.0 μg/mL. Cytotoxic tests using soursop seed protein with the concentrations of 32.2 μg/mL; 16.1 μg/mL; 8.05 μg/mL; and 4.025 μg/mL respectively resulted in 114.436%; 116.822%; 117.776%; and 119.526% HeLa cell survival. Meanwhile, isolated proteins with the concentration of 23.7 g/mL; 11.85 g/mL; 5.925 g/mL; and 2.9625 g/mL respectively resulted in 90.671%; 115.284%; 123.025%; and 127.590% of Vero cell survival. In conclusion, soursop seed protein did not possess cytotoxic activity against HeLa and Vero cells.
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