Highly sensitive and portable detection of PD-L1+ exosomes using a smartphone-assisted colorimetric sensor

Abstract

PD-L1⁺ exosomes act as a useful biomarker in early cancer diagnosis, therapeutic monitoring, prognostic assessment, and immunotherapy for non-invasive liquid biopsy. There exists urgent clinical need for developing rapid, portable, and cost-effective immediate-response assays for PD-L1⁺ exosomes. Here, we proposed a smartphone-assisted colorimetric sensor using the triple-helix molecular switch (THMS) combined with Y-shaped catalytic hairpin assembly (Y-CHA) reaction. Namely, this strategy initiates the Y-CHA cycle and forms Y-DNA G-quadruplex/Hemin DNAzyme with K⁺ and Hemin after specific recognition of PD-L1⁺ exosomes by THMS. Then, PD-L1⁺ exosomes could be quantified via the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB)-mediated color change. The method demonstrates excellent specificity and sensitivity, with a limit of detection (LOD) of 5.45 × 10³ particles/mL and an LOD of 8.56 × 10³ particles/mL using smartphone analysis. With this strategy, we found that PD-L1⁺ exosome levels were significantly elevated in the peripheral blood of patients with colorectal cancer liver metastases (CRLM), indicating great potential for identifying CRLM patients. Additionally, this colorimetric sensor can be used to quantify PD-L1⁺ exosomes across wide range of cancers, which possess a great prospect in the point-of-care testing filed for cancer companion diagnosis.