<p>Albumin, a highly biocompatible protein, has been utilized in the construction of anti-cancer drugs for the treatment of various cancers. However, albumin-based nanomedicines still face several clinical challenges, including low stability in the bloodstream, non-specific targeting ability, and lack of controlled release capability. To address these limitations, we developed a multiprotein-based nanomedicine for targeted chemotherapy of non-small cell lung cancer (NSCLC). This nanomedicine was assembled using a heart-shaped albumin and two Y-shaped anti-CD44 and anti-CD133 antibodies. To enhance stability and improve therapeutic efficacy, the multiproteins were crosslinked using disulfide bond linkers and loaded with paclitaxel and ceramide. This resulted in nanomedicines that exhibited responsiveness to glutathione (GSH) and demonstrated inhibition of tumor cancer cells and cancer stem cells (CSCs). Our experimental results indicated that the inclusion of anti-CD44 and anti-CD133 antibodies enhanced the targeting capability of nanomedicines towards cancer cells and CSCs in an in vitro study and the accumulation in both normoxic regions and hypoxic niches in in vivo tumor xenografts. The multiprotein-based nanomedicines also demonstrated GSH-dependent drug release behavior, induced apoptosis in cancer cells and CSCs, inhibited cell migration, and effectively suppressed NSCLC tumor growth. Overall, our findings presented a novel nanostructure created from differently shaped proteins for application in drug delivery. This multiprotein-based nanomedicines showed promising potential in addressing the limitations of albumin-based nanomedicines and may offer improved therapeutic outcomes for cancer treatment.</p> Graphical Abstract <p></p>