<p>Robust cell manufacturing demands minimally invasive methods to enrich and quantify representative secretome biomarkers, thereby providing critical insights into cellular phenotype and therapeutic potency. Here, we evaluated a titanium dioxide (TiO₂)-based batch adsorption platform for the non-invasive capture and quantification of secretome-associated biomolecules from human embryonic kidney 293T (HEK293T) and bone marrow-derived mesenchymal stem/stromal cell (MSC) cultures. Leveraging TiO₂’s affinity for phosphate-containing biomolecules, we first evaluated TiO₂ powders and microspheres using standardized green fluorescent protein (GFP)-labeled extracellular vesicles (EVs) to benchmark adsorption behavior, and subsequently applied the workflow to capture phosphatidylcholine (PC) and extracellular ribonucleic acid (exRNA) directly from conditioned media. Adsorption performance was systematically evaluated under serum-supplemented, serum-free, protein-supplemented, and preconditioned conditions, including ultracentrifuged secretome fractions. Preconditioning to remove serum, bovine serum albumin (BSA), and phenol red reduced background interference and improved biomarker capture (i.e., PC and exRNA). Under optimized conditions, TiO₂ enabled consistent PC capture, whereas exRNA recovery remained limited, likely reflecting weaker molecular interactions and competitive adsorption effects within complex secretomes. Collectively, these results delineate operational parameters enabling measurable biomarker recovery and demonstrate the bench-scale feasibility of TiO₂ as a practical, low-cost platform for secretome analysis. Overall, these findings lay the groundwork for TiO₂-based adsorption systems as non-invasive tools to support future real-time monitoring of cell culture dynamics in cell manufacturing.</p> Graphical abstract <p></p>