Aptamer-Cholesterol-Mediated Proximity Ligation Assay for Accurate Identification of Exosomes

Accurate identification of exosomes plays an essential role in facilitating disease diagnosis and therapies. Herein, we proposed an Aptamer-cholesterol-mediated Proximity Ligation Assay (AcmPLA) for accurate identification of exosomes in a dual-probe strategy, one aptamer probe for recognition of ex...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2020-04, Vol.92 (7), p.5411-5418
Main Authors: Zhao, Xianxian, Luo, Canjun, Mei, Qiang, Zhang, Hongmin, Zhang, Wenqing, Su, Dongwei, Fu, Weiling, Luo, Yang
Format: Article
Language:English
Subjects:
A549 Cells
Amplification
Analytical chemistry
Antibodies
Aptamers
Aptamers, Nucleotide - metabolism
Beads
Biomarkers
Biomedical engineering
Biosensing Techniques - methods
CD63 antigen
CD9 antigen
Chemistry
Cholesterol
Cholesterol - metabolism
Exosomes
Exosomes - metabolism
Humans
Identification
Limit of Detection
Molecular Probes - metabolism
Precision medicine
Proximity
Recognition
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Summary:Accurate identification of exosomes plays an essential role in facilitating disease diagnosis and therapies. Herein, we proposed an Aptamer-cholesterol-mediated Proximity Ligation Assay (AcmPLA) for accurate identification of exosomes in a dual-probe strategy, one aptamer probe for recognition of exosomal innate surface protein CD63 and another cholesterol probe for biolipid layer targeting. By integrating a proximity ligation of probes bound with exosomal biomarkers for specific recognition and a rolling circle amplification (RCA) strategy for signal amplification, we have successfully developed an exosomes-surface approach that can perform “AND” logic analysis of dual biomarkers, which not only could be used for exosomes quantification, but also for exosomes tracing. Besides RCA-initiated signal amplification, CD9 antibody-labeled magnetic beads were used to capture exosomes for isolation and secondary signal enrichment. Our approach can achieve specific exosomes isolation and accurate identification and thus could be exploited for broad applications in biological science, biomedical engineering, and personalized medicine.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.0c00141