Gold (Au) agglomerates (AGs) are reassembled using Triton X‐100 (T) and doxorubicin (D) dissolved in ethanol under 185 nm photoirradiation to form TAuD nanovesicles (NVs) under ambient gas flow conditions. The positively charged Au particles are then electrostatically conjugated with the anionic chains of TD components via a flowing drop (FD) reaction. Photoirradiation of the droplets in a tubular reactor continues the photophysicochemical reactions, resulting in the reassembly of Au AGs and TD into TAuD NVs. The fabricated NVs are electrostatically collected onto a polished aluminum rod in a single‐pass configuration. The dispersion of NVs is employed for bioassays to confirm uptake by cells and accumulation in tumors. The chemo‐photothermal activity is determined both in vitro and in vivo. Different combinations of components are also used to fabricate NVs using the FD reaction, and these NVs are suitable for gene delivery as well. This newly designed gaseous single‐pass process results in the reassembly of Au AGs for incorporation with TD without the need of batch wet chemical reactions, modifications, separations, or purifications. Thus, this process offers an efficient platform for preparing biofunctional Au nanostructures that requires neither complex physicochemical steps nor special storage techniques.