Cholesterol-DNA Labeling of Extracellular Vesicles for Amplification Quantitation

Patent Status

Brief Description

Quantifying specific subpopulations of extracellular vesicles is a critical challenge in liquid biopsy and disease monitoring. Researchers at UC Berkeley have developed a highly sensitive method that uses deoxyribonucleic acid oligonucleotide tagging to label and quantify these vesicles. The technology employs lipid-tagged single-stranded DNA that embeds itself directly into the lipid bilayer membrane of the vesicles. Through a combination of anchor, co-anchor, and detection oligonucleotides, a stable and programmable label is formed. Once specific subpopulations of vesicles are captured using antibody-antigen binding, the single-stranded DNA labels are released using restriction enzymes and measured via quantitative polymerase chain reaction. This approach provides a quantitative readout that is directly correlated with the number of captured vesicles, enabling the detection of specific biomarkers from complex biological fluids.

Suggested uses

• Disease Diagnosis and Staging: Detecting and monitoring conditions such as sepsis, traumatic brain injury, heart disease, and various cancers through blood or urine samples.

• Neurodegenerative Disease Monitoring: Tracking the progression of Alzheimer’s disease or dementia by quantifying specific neural-derived vesicles.

• Personalized Medicine: Evaluating an individual patient's response to cancer treatments by monitoring changes in vesicle subpopulations over time.

• Drug Discovery: Assaying the impact of new therapeutic compounds on cellular health and vesicle secretion in laboratory models.

• Multiplexed Biomarker Discovery: Simultaneously quantifying multiple different types of extracellular vesicles in a single sample to identify complex disease signatures.

Advantages

• High Sensitivity: Utilizing quantitative polymerase chain reaction allows for the detection of extremely low concentrations of vesicles that traditional methods might miss.

• Specific Subpopulation Targeting: The integration of immunocapture ensures that only the relevant vesicles expressing specific surface antigens are measured.

• Stable Membrane Anchoring: The use of self-embedding lipids and complementary DNA base pairing prevents the labels from dissociating during the capture and washing steps.

• Direct Correlation: The signal generated by the DNA amplification provides a precise numerical count that corresponds directly to the physical number of vesicles present.

• Versatile Sample Compatibility: The method is effective for vesicles isolated from a wide variety of sources, including blood, saliva, urine, and cell culture media.

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