Construction and application of a magnetic and catalytic hairpin assembly-based platform for detecting dual membrane proteins on exosomes / 南方医科大学学报
Journal of Southern Medical University
;
(12): 1453-1460, 2019.
Article
in Chinese
| WPRIM
| ID: wpr-781252
ABSTRACT
OBJECTIVE@#To construct a magnetic and catalytic hairpin assembly-based platform for detection of dual membrane proteins on exosomes.@*METHODS@#Exosomes in supernatant of breast cancer MDA-MB-231 cell culture were separated, purified and characterized. Super-resolution imaging and Western blotting were performed to confirm the expression of the membrane protein CD63 on the exosomes. Polyacrylamide gel electrophoresis was used to verify the combination of Apt-T and exosomes. Fluorescence experiments were carried out to test the feasibility of CHA nucleic acid sequence, optimize the reaction conditions, and determine the specificity of the detection platform.@*RESULTS@#Super-resolution imaging and Western blotting showed that breast cancer MDA-MB-231 cell-derived exosomes expressed abundant membrane protein CD63. Polyacrylamide gel electrophoresis indicated that Apt-T could recognize and bind to exosomes. The results of specificity test showed that the signal-to-noise ratio of the detection platform was 1.10±0.01 for detecting normal human breast epithelial cell-derived exosomes, and was 2.09±0.08 for breast cancer cell-derived exosomes.@*CONCLUSIONS@#Magnetic and catalytic hairpin assembly-based detection platform allows simultaneous detection of two membrane proteins expressed on exosomes and identification of the expressions of membrane proteins on exosomes from different sources.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Blotting, Western
/
Sensitivity and Specificity
/
Exosomes
/
Magnetic Phenomena
/
Membrane Proteins
Type of study:
Diagnostic study
Limits:
Humans
Language:
Chinese
Journal:
Journal of Southern Medical University
Year:
2019
Type:
Article
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