scholarly journals Isolation and characterisation of graves’ disease-specific extracellular vesicles from tissue maintained on a bespoke microfluidic device

2021 ◽  
pp. 100011
Author(s):  
Hayley Foster ◽  
Mark Wade ◽  
James England ◽  
John Greenman ◽  
Victoria Green
2005 ◽  
Vol 13 (11) ◽  
pp. 1223-1230 ◽  
Author(s):  
Bryan M Dechairo ◽  
Delilah Zabaneh ◽  
Joanne Collins ◽  
Oliver Brand ◽  
Gary J Dawson ◽  
...  

Micromachines ◽  
2021 ◽  
Vol 12 (12) ◽  
pp. 1534
Author(s):  
Junyuan Liu ◽  
Yuxin Qu ◽  
Han Wang

Methods for the isolation and analysis of extracellular vesicles (EVs) have been extensively explored in the field of life science and in clinical diagnosis in recent years. The separation and efficient recovery of high-purity target EVs from biological samples are important prerequisites in the study of EVs. So far, commonly used methods of EV separation include ultracentrifugation, filtration, solvent precipitation and immunoaffinity capturing. However, these methods suffer from long processing time, EV damage and low enrichment efficiency. The use of acoustophoretic force facilitates the non-contact label-free manipulation of cells based on their size and compressibility but lacks specificity. Additionally, the acoustophoretic force exerted on sub-micron substances is normally weak and insufficient for separation. Here we present a novel immuno-acoustic sorting technology, where biological substances such as EVs, viruses, and biomolecules, can be specifically captured by antibody/receptor coated microparticles through immunoaffinity, and manipulated by an acoustophoretic force exerted on the microparticles. Using immuno-acoustic sorting technology, we successfully separated and purified HER2-positive EVs for further downstream analysis. This method holds great potential in isolating and purifying specific targets such as disease-related EVs from biological fluids and opens new possibilities for the EV-based early diagnosis and prognosis of diseases.


Cancers ◽  
2021 ◽  
Vol 13 (19) ◽  
pp. 4968
Author(s):  
Martina Barone ◽  
Monica Barone ◽  
Francesca Ricci ◽  
Giuseppe Auteri ◽  
Francesco Fabbri ◽  
...  

Polycythemia vera is a myeloproliferative neoplasm with increased risk of thrombosis and progression to myelofibrosis. However, no disease-specific risk factors have been identified so far. Circulating extracellular vesicles (EVs) are mostly of megakaryocyte (MK-EVs) and platelet (PLT-EVs) origin and, along with phosphatidylethanolamine (PE)-EVs, play a role in cancer and thrombosis. Interestingly, circulating microbial components/microbes have been recently indicated as potential modifiers of inflammation and coagulation. Here, we investigated phenotype and microbial DNA cargo of EVs after isolation from the plasma of 38 patients with polycythemia vera. Increased proportion of MK-EVs and reduced proportion of PLT-EVs identify patients with thrombosis history. Interestingly, EVs from patients with thrombosis history were depleted in Staphylococcus DNA but enriched in DNA from Actinobacteria members as well as Anaerococcus. In addition, patients with thrombosis history had also lower levels of lipopolysaccharide-associated EVs. In regard to fibrosis, along with increased proportion of PE-EVs, the EVs of patients with marrow fibrosis were enriched in DNA from Collinsella and Flavobacterium. Here, we identified a polycythemia-vera-specific host/microbial EV-based signature associated to thrombosis history and marrow fibrosis. These data may contribute to refining PV prognosis and to identifying novel druggable targets.


The Analyst ◽  
2019 ◽  
Vol 144 (19) ◽  
pp. 5785-5793 ◽  
Author(s):  
Yoon-Tae Kang ◽  
Emma Purcell ◽  
Thomas Hadlock ◽  
Ting-Wen Lo ◽  
Anusha Mutukuri ◽  
...  

We present a simple strategy to immobilize and analyze extracellular vesicles for multiple markers on a microfluidic device, called DICE.


2015 ◽  
Vol 54 (03) ◽  
pp. 131-136 ◽  
Author(s):  
F. F. Behrendt ◽  
A. Heinzel ◽  
B. Müller ◽  
F. M. Mottaghy ◽  
F. A. Verburg ◽  
...  

SummaryAim: To compare uptake measurements and different methods for the pretherapeutic determination of the effective thyroidal 131I half life (Teff) to the results of posttherapeutic dosimetric measurements. Patients, methods: Retrospective study of 1538 patients who received their first RIT in our department for autonomous thyroid nodules (ATN), autonomous multinodular goiter (AMG) or Graves' disease (GD) between November 1999 and January 2011. Pretherapeutic measurements were performed at any combination of 24 h, 48 h and 6 days after 131I administration. Post-therapy dosimetric measurements were performed in 12 h intervals until discharge. Teff was determined through monoexponential curve fitting. Results: Pretherapeutic Teff values based on measurements at 24 h and 48 h, 24 h and 6 d, 48 h and 6 d as well as on day 24 h, 48 h and 6 d yielded implausible (< 2 d or > 8 d) values for Teff, in 60.4%, 25.7%, 29.1 and 21.4% of available calculations, respectively. The plausible results showed significant, clinically relevant and sometimes considerable overestimations of Teff. Using empirically determined fixed disease specific Teff values resulted in a better congruence between the pre- and posttherapeutic dosimetry results. 24 h measurements were marginally more accurate than 48 h ones in AMG and GD whereas 48 h measurements were marginally more accurate in ATN; these differences are however not clinically relevant. 6 d measurements are clearly less accurate than those after 24 h or 48 h. Conclusion: In ATN, AMG and GD, pretherapeutic dosimetry can be performed by a single uptake measurement at 24 h or 48 h using a fixed, disease specific value for Teff. Additional later measurements do not yield a further clinically relevant contribution to accuracy of pretherapeutic dosimetry.


2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Harshani Wijerathne ◽  
Malgorzata A. Witek ◽  
Joshua M. Jackson ◽  
Virginia Brown ◽  
Mateusz L. Hupert ◽  
...  

Abstract Currently there is no in vitro diagnostic test for acute ischemic stroke (AIS), yet rapid diagnosis is crucial for effective thrombolytic treatment. We previously demonstrated the utility of CD8(+) T-cells’ mRNA expression for AIS detection; however extracellular vesicles (EVs) were not evaluated as a source of mRNA for AIS testing. We now report a microfluidic device for the rapid and efficient affinity-enrichment of CD8(+) EVs and subsequent EV’s mRNA analysis using droplet digital PCR (ddPCR). The microfluidic device contains a dense array of micropillars modified with anti-CD8α monoclonal antibodies that enriched 158 ± 10 nm sized EVs at 4.3 ± 2.1 × 109 particles/100 µL of plasma. Analysis of mRNA from CD8(+) EVs and their parental T-cells revealed correlation in the expression for AIS-specific genes in both cell lines and healthy donors. In a blinded study, 80% test positivity for AIS patients and controls was revealed with a total analysis time of 3.7 h.


Lab on a Chip ◽  
2020 ◽  
Vol 20 (10) ◽  
pp. 1762-1770 ◽  
Author(s):  
Ting-Wen Lo ◽  
Ziwen Zhu ◽  
Emma Purcell ◽  
Daniel Watza ◽  
Joyful Wang ◽  
...  

Immunoaffinity based EV isolation technologies use antibodies targeting surface markers on EVs to provide higher isolation specificity and purity compared to existing approaches.


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