Colorimetric Aptasensor Coupled with a Deep-learning-powered Smartphone App for Programmed Death Ligand-1 Expressing Extracellular Vesicles

Overview

Journal Front Immunol

Date 2025 Feb 7

PMID 39916963

Authors

Adeel Khan

Haroon Khan

Nongyue He

Zhiyang Li

Heba Khalil Alyahya

Yousef A Bin Jardan

Affiliations

Soon will be listed here.

Abstract

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 (FeO)-SiO-TiO 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 3.6×10 EVs/mL with a wide linear range from 10 to 10 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.

References

Kang W, Liu L, Yu P, Zhang T, Lei C, Nie Z . A switchable Cas12a enabling CRISPR-based direct histone deacetylase activity detection. Biosens Bioelectron. 2022; 213:114468. DOI: 10.1016/j.bios.2022.114468. View

Boriachek K, Masud M, Palma C, Phan H, Yamauchi Y, Hossain M . Avoiding Pre-Isolation Step in Exosome Analysis: Direct Isolation and Sensitive Detection of Exosomes Using Gold-Loaded Nanoporous Ferric Oxide Nanozymes. Anal Chem. 2019; 91(6):3827-3834. DOI: 10.1021/acs.analchem.8b03619. View

Niu M, Liu Y, Yi M, Jiao D, Wu K . Biological Characteristics and Clinical Significance of Soluble PD-1/PD-L1 and Exosomal PD-L1 in Cancer. Front Immunol. 2022; 13:827921. PMC: 8977417. DOI: 10.3389/fimmu.2022.827921. View

Xie S, Tao D, Fu Y, Xu B, Tang Y, Steinaa L . Rapid Visual CRISPR Assay: A Naked-Eye Colorimetric Detection Method for Nucleic Acids Based on CRISPR/Cas12a and a Convolutional Neural Network. ACS Synth Biol. 2021; 11(1):383-396. DOI: 10.1021/acssynbio.1c00474. View

Huang M, Yang J, Wang T, Song J, Xia J, Wu L . Homogeneous, Low-volume, Efficient, and Sensitive Quantitation of Circulating Exosomal PD-L1 for Cancer Diagnosis and Immunotherapy Response Prediction. Angew Chem Int Ed Engl. 2020; 59(12):4800-4805. DOI: 10.1002/anie.201916039. View

Chang L, Wu H, Chen R, Sun X, Yang Y, Huang C . Microporous PdCuB nanotag-based electrochemical aptasensor with Au@CuCl nanowires interface for ultrasensitive detection of PD-L1-positive exosomes in the serum of lung cancer patients. J Nanobiotechnology. 2023; 21(1):86. PMC: 10008610. DOI: 10.1186/s12951-023-01845-y. View

Xia N, Cheng J, Tian L, Zhang S, Wang Y, Li G . Hybridization Chain Reaction-Based Electrochemical Biosensors by Integrating the Advantages of Homogeneous Reaction and Heterogeneous Detection. Biosensors (Basel). 2023; 13(5). PMC: 10216504. DOI: 10.3390/bios13050543. View

Khan A, Di K, Khan H, He N, Li Z . Rapid Capturing and Chemiluminescent Sensing of Programmed Death Ligand-1 Expressing Extracellular Vesicles. Biosensors (Basel). 2022; 12(5). PMC: 9138745. DOI: 10.3390/bios12050281. View

Li J, Baird M, Davis M, Tai W, Zweifel L, Adams Waldorf K . Dramatic enhancement of the detection limits of bioassays via ultrafast deposition of polydopamine. Nat Biomed Eng. 2017; 1. PMC: 5654575. DOI: 10.1038/s41551-017-0082. View

10.

Wang Y, Liu J, Adkins G, Shen W, Trinh M, Duan L . Enhancement of the Intrinsic Peroxidase-Like Activity of Graphitic Carbon Nitride Nanosheets by ssDNAs and Its Application for Detection of Exosomes. Anal Chem. 2017; 89(22):12327-12333. PMC: 5705088. DOI: 10.1021/acs.analchem.7b03335. View

11.

Sun B, Zhou A, Li X, Yu H . Development and Application of Mobile Apps for Molecular Sensing: A Review. ACS Sens. 2021; 6(5):1731-1744. DOI: 10.1021/acssensors.1c00512. View

12.

Wang Y, Liu Z, Wang X, Dai Y, Li X, Gao S . Rapid and Quantitative Analysis of Exosomes by a Chemiluminescence Immunoassay Using Superparamagnetic Iron Oxide Particles. J Biomed Nanotechnol. 2019; 15(8):1792-1800. DOI: 10.1166/jbn.2019.2809. View

13.

Zhao J, Li X, Liu L, Zhu Z, He C . Exosomes in lung cancer metastasis, diagnosis, and immunologically relevant advances. Front Immunol. 2023; 14:1326667. PMC: 10752943. DOI: 10.3389/fimmu.2023.1326667. View

14.

Cao Y, Wang Y, Yu X, Jiang X, Li G, Zhao J . Identification of programmed death ligand-1 positive exosomes in breast cancer based on DNA amplification-responsive metal-organic frameworks. Biosens Bioelectron. 2020; 166:112452. DOI: 10.1016/j.bios.2020.112452. View

15.

Yu Q, Chen X, Qi L, Yang H, Wang Y, Zhang M . Smartphone readable colorimetry and ICP-MS dual-mode sensing platform for ultrasensitive and label-free detection of Escherichia coli based on filter-assisted separation. Talanta. 2022; 251:123760. DOI: 10.1016/j.talanta.2022.123760. View

16.

Chai H, Cheng W, Jin D, Miao P . Recent Progress in DNA Hybridization Chain Reaction Strategies for Amplified Biosensing. ACS Appl Mater Interfaces. 2021; 13(33):38931-38946. DOI: 10.1021/acsami.1c09000. View

17.

Su X, Liu X, Xie Y, Chen M, Zheng C, Zhong H . Integrated SERS-Vertical Flow Biosensor Enabling Multiplexed Quantitative Profiling of Serological Exosomal Proteins in Patients for Accurate Breast Cancer Subtyping. ACS Nano. 2023; 17(4):4077-4088. DOI: 10.1021/acsnano.3c00449. View

18.

Gao F, Jiao F, Xia C, Zhao Y, Ying W, Xie Y . A novel strategy for facile serum exosome isolation based on specific interactions between phospholipid bilayers and TiO. Chem Sci. 2019; 10(6):1579-1588. PMC: 6369439. DOI: 10.1039/c8sc04197k. View

19.

Liu D, Tang J, Xu H, Yuan K, Aryee A, Zhang C . Split-aptamer mediated regenerable temperature-sensitive electrochemical biosensor for the detection of tumour exosomes. Anal Chim Acta. 2022; 1219:340027. DOI: 10.1016/j.aca.2022.340027. View

20.

Wang L, Yang Y, Liu Y, Ning L, Xiang Y, Li G . Bridging exosome and liposome through zirconium-phosphate coordination chemistry: a new method for exosome detection. Chem Commun (Camb). 2019; 55(18):2708-2711. DOI: 10.1039/c9cc00220k. View