The present study focused on the extraction, purification, and characterization of proteins from seeds of Vitis vinifera and Mangifera indica, along with evaluation of their anti-proliferative potential. Protein estimation using the Lowry method revealed higher protein content in grape seed extract (4.98 ± 0.14 mg/mL) compared to mango seed extract (4.20 ± 0.12 mg/mL). Spectrophotometric analysis at 280 nm showed higher absorbance for grape seed protein (1.32 ± 0.05) than mango seed protein (1.08 ± 0.04), confirming greater protein abundance. SDS-PAGE profiling indicated multiple protein bands in the range of 30–97 kDa for grape seeds and 35–80 kDa for mango seeds, with grape seed proteins exhibiting greater molecular diversity. The anti-proliferative activity of the purified proteins was assessed using the MTT assay against HeLa and MCF-7 cancer cell lines. A dose-dependent decrease in cell viability was observed, with HeLa cells showing higher sensitivity (IC₅₀ ≈ 120 µg/mL) compared to MCF-7 cells (IC₅₀ ≈ 135 µg/mL). At 200 µg/mL, cell viability decreased to 29.5% in HeLa and 35.0% in MCF-7 cells. Morphological assessment uses light microscopy and acridine orange/ethidium bromide staining revealed characteristic apoptotic features, including cell shrinkage, membrane blebbing, chromatin condensation, and nuclear fragmentation, confirming apoptosis as the primary mechanism of cell death. Overall, the results indicate that grape seed proteins possess higher protein content, greater structural complexity, and stronger anti-proliferative activity compared to mango seed proteins. These findings highlight the potential of seed-derived proteins as promising natural candidates for anticancer applications and support further investigation into their molecular mechanisms.
