scholarly journals Rapid Detection and Trapping of Extracellular Vesicles by Electrokinetic Concentration for Liquid Biopsy on Chip

Micromachines ◽  
2018 ◽  
Vol 9 (6) ◽  
pp. 306 ◽  
Author(s):  
Lucia Cheung ◽  
Sarah Sahloul ◽  
Ajymurat Orozaliev ◽  
Yong-Ak Song
Keyword(s):  
Extracellular Vesicles ◽  
Liquid Biopsy ◽  
Rapid Detection ◽  
On Chip

scholarly journals Rapid detection of exosomal microRNA biomarkers by electrokinetic concentration for liquid biopsy on chip

2018 ◽  
Vol 12 (1) ◽  
pp. 014104 ◽  
Author(s):  
Lucia S. Cheung ◽  
Xi Wei ◽  
Diogo Martins ◽  
Yong-Ak Song
Keyword(s):  
Liquid Biopsy ◽  
Rapid Detection ◽  
On Chip ◽  
Exosomal Microrna

scholarly journals Application of Microfluidic Chips in Separation and Analysis of Extracellular Vesicles in Liquid Biopsy for Cancer

Micromachines ◽  
2019 ◽  
Vol 10 (6) ◽  
pp. 390 ◽  
Author(s):  
Jin Lu ◽  
Jiushen Pang ◽  
Ying Chen ◽  
Qi Dong ◽  
Jiahao Sheng ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Liquid Biopsy ◽  
Culture Medium ◽  
Microfluidic Chips ◽  
Cell Culture Medium ◽  
Single Chip ◽  
Size Dependent ◽  
On Chip ◽  
Monitoring Treatment ◽  
Chip Analysis

Extracellular vesicles (EVs) are becoming a promising biomarker in liquid biopsy of cancer. Separation EV from cell culture medium or biofluids with high purity and quality remains a technique challenge. EV manipulation techniques based on microfluidics have been developed in the last decade. Microfluidic-based EV separation techniques developed so far can be classified into two categories: surface biomarker-dependent and size-dependent approaches. Microfluidic techniques allow the integration of EV separation and analysis on a single chip. Integrated EV separation and on-chip analysis have shown great potential in cancer diagnosis and monitoring treatment of responses. In this review, we discuss the development of microfluidic chips for EV separation and analysis. We also detail the clinical application of these microfluidic chips in the liquid biopsy of various cancers.

scholarly journals Single Step In Situ Detection of Surface Protein and MicroRNA in Clustered Extracellular Vesicles Using Flow Cytometry

2021 ◽  
Vol 10 (2) ◽  
pp. 319
Author(s):  
Hee Cheol Yang ◽  
Won Jong Rhee
Keyword(s):  
Extracellular Vesicles ◽  
Liquid Biopsy ◽  
Molecular Beacon ◽  
Disease Diagnosis ◽  
Single Step ◽  
Flow Cytometer ◽  
Surface Proteins ◽  
Biomarker Detection ◽  
In Situ Detection

Because cancers are heterogeneous, it is evident that multiplexed detection is required to achieve disease diagnosis with high accuracy and specificity. Extracellular vesicles (EVs) have been a subject of great interest as sources of novel biomarkers for cancer liquid biopsy. However, EVs are nano-sized particles that are difficult to handle; thus, it is necessary to develop a method that enables efficient and straightforward EV biomarker detection. In the present study, we developed a method for single step in situ detection of EV surface proteins and inner miRNAs simultaneously using a flow cytometer. CD63 antibody and molecular beacon-21 were investigated for multiplexed biomarker detection in normal and cancer EVs. A phospholipid-polymer-phospholipid conjugate was introduced to induce clustering of the EVs analyzed using nanoparticle tracking analysis, which enhanced the detection signals. As a result, the method could detect and distinguish cancer cell-derived EVs using a flow cytometer. Thus, single step in situ detection of multiple EV biomarkers using a flow cytometer can be applied as a simple, labor- and time-saving, non-invasive liquid biopsy for the diagnosis of various diseases, including cancer.

scholarly journals A multiparametric extraction method for Vn96-isolated plasma extracellular vesicles and cell-free DNA that enables multi-omic profiling

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jeremy W. Roy ◽  
Catherine A. Taylor ◽  
Annie P. Beauregard ◽  
Surendar R. Dhadi ◽  
D. Craig Ayre ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Liquid Biopsy ◽  
Small Rna Sequencing ◽  
Isolation Method ◽  
Protein Biomarkers ◽  
Cell Free Dna ◽  
Extraction Protocol ◽  
Free Dna ◽  
Rich Material ◽  
Short Timeframe

AbstractExtracellular vesicles (EVs) have been recognized as a rich material for the analysis of DNA, RNA, and protein biomarkers. A remaining challenge for the deployment of EV-based diagnostic and prognostic assays in liquid biopsy testing is the development of an EV isolation method that is amenable to a clinical diagnostic lab setting and is compatible with multiple types of biomarker analyses. We have previously designed a synthetic peptide, known as Vn96 (ME kit), which efficiently isolates EVs from multiple biofluids in a short timeframe without the use of specialized lab equipment. Moreover, it has recently been shown that Vn96 also facilitates the co-isolation of cell-free DNA (cfDNA) along with EVs. Herein we describe an optimized method for Vn96 affinity-based EV and cfDNA isolation from plasma samples and have developed a multiparametric extraction protocol for the sequential isolation of DNA, RNA, and protein from the same plasma EV and cfDNA sample. We are able to isolate sufficient material by the multiparametric extraction protocol for use in downstream analyses, including ddPCR (DNA) and ‘omic profiling by both small RNA sequencing (RNA) and mass spectrometry (protein), from a minimum volume (4 mL) of plasma. This multiparametric extraction protocol should improve the ability to analyse multiple biomarker materials (DNA, RNA and protein) from the same limited starting material, which may improve the sensitivity and specificity of liquid biopsy tests that exploit EV-based and cfDNA biomarkers for disease detection and monitoring.

scholarly journals Peptide-Affinity Precipitation of Extracellular Vesicles and Cell-Free DNA Improves Sequencing Performance for the Detection of Pathogenic Mutations in Lung Cancer Patient Plasma

2020 ◽  
Vol 21 (23) ◽  
pp. 9083
Author(s):  
Catherine Taylor ◽  
Simi Chacko ◽  
Michelle Davey ◽  
Jacynthe Lacroix ◽  
Alexander MacPherson ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Extracellular Vesicles ◽  
Liquid Biopsy ◽  
Nsclc Patient ◽  
Egfr Mutations ◽  
Single Nucleotide Variants ◽  
Cell Free Dna ◽  
Free Dna ◽  
Nsclc Patients ◽  
Peptide Affinity

Liquid biopsy is a minimally-invasive diagnostic method that may improve access to molecular profiling for non-small cell lung cancer (NSCLC) patients. Although cell-free DNA (cf-DNA) isolation from plasma is the standard liquid biopsy method for detecting DNA mutations in cancer patients, the sensitivity can be highly variable. Vn96 is a peptide with an affinity for both extracellular vesicles (EVs) and circulating cf-DNA. In this study, we evaluated whether peptide-affinity (PA) precipitation of EVs and cf-DNA from NSCLC patient plasma improves the sensitivity of single nucleotide variants (SNVs) detection and compared observed SNVs with those reported in the matched tissue biopsy. NSCLC patient plasma was subjected to either PA precipitation or cell-free methods and total nucleic acid (TNA) was extracted; SNVs were then detected by next-generation sequencing (NGS). PA led to increased recovery of DNA as well as an improvement in NGS sequencing parameters when compared to cf-TNA. Reduced concordance with tissue was observed in PA-TNA (62%) compared to cf-TNA (81%), mainly due to identification of SNVs in PA-TNA that were not observed in tissue. EGFR mutations were detected in PA-TNA with 83% sensitivity and 100% specificity. In conclusion, PA-TNA may improve the detection limits of low-abundance alleles using NGS.

scholarly journals On-Chip Liquid Biopsy: Progress in Isolation of Exosomes for Early Diagnosis of Cancer

2017 ◽  
Vol 112 (3) ◽  
pp. 461a ◽  
Author(s):  
Sung Cheol Kim ◽  
Navneet Dogra ◽  
Benjamin H. Wunsch ◽  
Joshua T. Smith ◽  
Stacey M. Gifford ◽  
...  
Keyword(s):  
Early Diagnosis ◽  
Liquid Biopsy ◽  
Early Diagnosis Of Cancer ◽  
Diagnosis Of Cancer ◽  
On Chip

Separation of distinct exosome subpopulations: isolation and characterization approaches and their associated challenges

The Analyst ◽  
2021 ◽  
Author(s):  
Karishma Singh ◽  
Ruchika Nalabotala ◽  
Kevin M. Koo ◽  
Sudeep Bose ◽  
Ranu Nayak ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Liquid Biopsy ◽  
Therapeutic Use ◽  
Isolation And Characterization ◽  
Communications System

Exosomes are nano-sized extracellular vesicles that serve as a communications system between cells and has shown tremendous promise as liquid biopsy biomarkers in diagnostic, prognostic, and even therapeutic use in...

Protein biomarkers in breast cancer-derived extracellular vesicles for use in liquid biopsies

2021 ◽  
Author(s):  
Dan Li ◽  
Wenjia Lai ◽  
Di Fan ◽  
Qiaojun Fang
Keyword(s):  
Breast Cancer ◽  
Early Diagnosis ◽  
Extracellular Vesicles ◽  
Liquid Biopsy ◽  
Breast Cancer Diagnosis ◽  
Extracellular Vesicle ◽  
Detection Methods ◽  
Protein Markers ◽  
Liquid Biopsies ◽  
Diagnosis And Prognosis

Breast cancer is the most common malignant disease in women worldwide. Early diagnosis and treatment can greatly improve the management of breast cancer. Liquid biopsies are becoming convenient detection methods for diagnosing and monitoring breast cancer due to their non-invasiveness and ability to provide real-time feedback. A range of liquid biopsy markers, including circulating tumor proteins, circulating tumor cells, and circulating tumor nucleic acids, have been implemented for breast cancer diagnosis and prognosis, with each having its own advantages and limitations. Circulating extracellular vesicles are messengers of intercellular communication that are packed with information from mother cells and are found in a wide variety of bodily fluids; thus, they are emerging as ideal candidates for liquid biopsy biomarkers. In this review, we summarize extracellular vesicle protein markers that can be potentially used for the early diagnosis and prognosis of breast cancer or determining its specific subtypes.

scholarly journals Enhanced recovery of CD9-positive extracellular vesicles from human specimens by chelating reagent

2020 ◽  
Author(s):  
Ayako Kurimoto ◽  
Yuki Kawasaki ◽  
Toshiki Ueda ◽  
Tatsutoshi Inuzuka
Keyword(s):  
Extracellular Vesicles ◽  
Western Blotting ◽  
Biological Samples ◽  
Liquid Biopsy ◽  
Enhanced Recovery ◽  
Recovery Efficiency ◽  
Early Diagnostics ◽  
Isolation Methods ◽  
Chelating Reagent

AbstractExtracellular vesicles (EVs) have gained attention as potential targets of early diagnostics and prognosis in the field of liquid biopsy. Despite clinical potentials, the best method to isolate EVs from specimens remains controversial due to low purity, low specificity, and lack of reproducibility with current isolation methods. Here we show that a chelating reagent enhances the recovery efficiency of EVs from crude biological samples by immunoprecipitation using an anti-CD9 antibody. Proteomic and western blotting analyses show that the EVs isolated using the chelating reagent contain a wider variety of proteins than those isolated with PBS.

An integrated microfluidic system for on-chip enrichment and quantification of circulating extracellular vesicles from whole blood

Lab on a Chip ◽  
2019 ◽  
Vol 19 (19) ◽  
pp. 3305-3315 ◽  
Author(s):  
Yi-Sin Chen ◽  
Yu-Dong Ma ◽  
Chihchen Chen ◽  
Shu-Chu Shiesh ◽  
Gwo-Bin Lee
Keyword(s):  
Extracellular Vesicles ◽  
Immunosorbent Assay ◽  
Whole Blood ◽  
Magnetic Beads ◽  
Extracellular Vesicle ◽  
Microfluidic System ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Whole Blood ◽  
On Chip ◽  
Chip Enrichment

An integrated microfluidic system was developed for extracellular vesicle (EV) enrichment and quantification by using anti-CD63-coated magnetic beads and an on-chip enzyme-linked immunosorbent assay in human whole blood.

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