Chemo-photodynamic therapy is an attractive strategy against cancer. However, the combination of chemotherapeutic agent and photosensitizer is usually realized by physically embedding one or two agents in the carriers. In this case, batch to batch variation of drug contents increases the risk of composition uncertainty, limiting the clinical application of currently developed chemo-photodynamic therapy. Therefore, it is highly desirable for constructing chemo-photodynamic therapy nanoplatform with precisely controllable composition and drug loading content. Here, a precise chemo-photodynamic therapy nanoplatform is reported based on the chemical conjugation of both doxorubicin (DOX) and photosensitizer chlorin e6 (Ce6) to active targeting hyaluronic acid (HA). The achieved HA-DOX-Ce6 (HDC) conjugate can self-assemble into all-in-one nanoparticles (NPs) with properties of pH-response, self-targeting delivery, near-infrared (NIR) fluorescence imaging and chemo-photodynamic combination therapy. Because HDC conjugate holds precise components, the HDC NPs have a constant composition and drug loading content, endowing a precise chemo-photodynamic therapy nanoplatform. The HDC NPs show significantly higher cellular uptake and remarkably better tumor-targeted accumulation than free drugs. Moreover, HDC NPs with the laser irradiation exhibit enhanced anticancer activities both in vitro and in vivo. This work may open the door for design of all-in-one advanced nanoplatform toward chemo-photodynamic therapy with precisely controllable composition and drug loading content. (C) 2019 Elsevier Inc. All rights reserved.