Data Sheet 1_Colorimetric aptasensor coupled with a deep-learning-powered smartphone app for programmed death ligand-1 expressing extracellular vesicles.docx

Lung cancer is a devastating public health threat and a leading cause of cancer-related deaths. Therefore, it is imperative to develop sophisticated techniques for the non-invasive detection of lung cancer. Extracellular vesicles expressing programmed death ligand-1 (PD-L1) markers (PD-L1@EVs) in the blood are reported to be indicative of lung cancer and response to immunotherapy. Our approach is the development of a colorimetric aptasensor by combining the rapid capturing efficiency of (Fe3O4)-SiO2-TiO2 for EV isolation with PD-L1 aptamer-triggered enzyme-linked hybridization chain reaction (HCR) for signal amplification. The numerous HRPs catalyze their substrate dopamine (colorless) into polydopamine (blackish brown). Change in chromaticity directly correlates with the concentration of PD-L1@EVs in the sample. The colorimetric aptasensor was able to detect PD-L1@EVs at concentrations as low as 3.6×102 EVs/mL with a wide linear range from 103 to 1010 EVs/mL with high specificity and successfully detected lung cancer patients’ serum from healthy volunteers’ serum. To transform the qualitative colorimetric approach into a quantitative operation, we developed an intelligent convolutional neural network (CNN)-powered quantitative analyzer for chromaticity in the form of a smartphone app named ExoP, thereby achieving the intelligent analysis of chromaticity with minimal user intervention or additional hardware attachments for the sensitive and specific quantification of PD-L1@EVs. This combined approach offers a simple, sensitive, and specific tool for lung cancer detection using PD-L1@EVs. The addition of a CNN-powered smartphone app further eliminates the need for specialized equipment, making the colorimetric aptasensor more accessible for low-resource settings.