ResearchDx is now offering EXO-NET® Exosome Isolation and Analysis services through a licensing agreement with Inoviq

EXO-NET® Pan-Exosome Capture is an immunoaffinity magnetic bead-based isolation system for Exosome Isolation and Analysis from plasma, serum, urine, saliva, and cell-conditioned medium.

EXO-NET®  offers unparalleled recovery of enriched exosomes for a multitude of applications.

 

 

Exosome Services offered by ResearchDx

High Throughput Exosome Extraction

  • DNA
  • RNA
  • MiRNA
  • Protein
 Analysis of EV associated miRNA

 

Exosome Analysis Services Offered By ResearchDx

  • qPCR Expression Analysis
  • miRNA NGS expression analysis
  • Total RNA NGS expression analysis
  • ELISA-based protein analysis
  • Fluorescence-activated Cell Sorting (FACS)
  • Flow Cytometry
Western blot analysis of EV

How Does EXO-NET® work?

EXO-NET is a fast and efficient exosome isolation system suitable for almost any biofluid or application. The EXO-NET workflow can be applied to high throughput screening of EVs and exosome-based biomarker discovery for diagnostics and therapeutic targets.

EXO-NET Performance

Comparison of mRNA and microRNA yield and recovery from EXO-NET isolated EVs with 4 other EV isolation kits.

Method: EVs were isolated from pooled normal human plasma by EXO-NET and other 4 commercial kits according to their manufacturers’ instructions.  Isolated EVs were lysed and total RNA was extracted for qPCR analysis to measure both microRNAs (miR-16, let-7a and miR-21) and mRNAs (GAPDH, OAZ1, RPLPO and SERF2).

Result: EXO-NET delivers equivalent or higher recovery of plasma EV RNA (mRNAs and microRNAs) compared to the other 4 commercial EV isolation kits as indicated by a lower CT value.  

 EV Associated miRNA

 

  

Evaluation of known EV protein and RNA markers in EXO-NET isolated EVs from human plasma.

Method: Pooled normal human plasma (500µl) was incubated with 30 µl of EXO-NET for 15 min. EXO-NET-captured EVs were magnetically isolated and lysed for downstream analysis. Total EV protein and RNA were extracted and analysed by Western blot and qPCR.

Result: Western blot analysis showed EXO-NET-isolated EVs from normal human plasma were positive for CD63, CD81, CD9, TSG101 and Flotillin-1, and negative for calnexin (an EV negative marker). qPCR analysis demonstrated that EXO-NET isolated EVs had high yield of mRNA (GAPDH) and microRNA (miR-191).

 Western Blot Analysis of EV's

 

  

Comparison of known EV proteins and RNA markers in EXO-NET isolated EVs from human saliva.

Method: Pooled normal human saliva (400µl) was diluted with 400µl of PBS and then incubated with 30 µl  of EXO-NET for 15 min.  EXO-NET captured EVs were magnetically isolated and lysed for downstream analysis.  Total protein and RNA were extracted from captured EVs and analyzed by Western blot and qPCR.

Result: Western blot analysis confirmed the presence of CD63, CD81 and CD9. Calnexin (EVs negative marker) was not detectable. qPCR analysis confirmed that EXO-NET isolated EVs had a high yield of mRNA (GAPDH) and microRNA (U6).

 Western Blot Analysis of EV's

 

  

Comparison of mass spectrometry proteomic analysis of EXO-NET isolated EVs with another bead-based EV isolation kit.

Method: EVs were isolated from pooled normal human plasma by EXO-NET and Kit A according to manufacturers’ instructions.  Isolated EVs were lysed and extracted total protein were processed for proteomic analysis by tandem mass spectrometry.

Result: Mass spectrometry analysis showed that albumin peptide intensity in Kit A was 2-fold higher than that observed for EXO-NET isolated EVs.  EXO-NET reduces contamination of EV preparations by serum proteins that may confound downstream analysis. The EV peptides intensity in Kit A was 6 to 10-fold less than that observed for EXO-NET-isolated EVs.

 EXO-NET isolated EV's
   

EXO-NET comparison to traditional exosome isolation methods1

EV Enrichment Techniques Time Cost Scalability Contaminants Specificity
Polyethylene glycol precipitation          
Size exclusion chromatography          
High MW centrifugal filters          
Differential ultracentrifugation          
Tangential flow filtration          
Affinity chromatography          
Immuno-magnetic bead capture*          

1Adapted from Salomon et al (2022) Extracellular Vesicles and Their Emerging Roles as Cellular Messengers in Endocrinology: An Endocrine Society Scientific Statement, Endocrine Reviews, Endocrine Reviews, 2022, 43, 441–468

 

Peer Review

Posters

  1. Poster Presentation: Thomas Stewart, Tamkin Ahmadzada, Sophia Amerena-Cowie, Emily Stein, Peter French, Gregory Rice, Michelle Wood, Joonsup Lee, Elham Hosseini-Beheshti, Peter A Lay, Georges E Grau. Differential detection of cancer-derived extracellular vesicles using combined antibody functionalized magnetic beads and infrared spectroscopy. 
  2. Poster Presentation: Andrew Lai, Dominic Guanzon, Carlos Palma, Ramin Khanabdali, Lewis Perrin, John Hooper, Jim Coward, Gregory Rice, Carlos Salomon. Reproducibility and efficiency of an extracellular vesicle capture technology for the detection of ovarian cancer. Presented at ISEV Conference in Seattle, USA (18 May 2023).
  3. Poster Presentation: Olesia Gololobova, Gregory Rice, Ramin Khanabdali, Kenneth W. Witwer. 
    Enrichment of extracellular vesicles from human plasma with molecular net matrix coated magnetic beads (EXO-NET®). Presented at ISEV Conference in Seattle, USA (19 May 2023).
  4. Poster Presentation: Ramin Khanabdali, Carlos Palma, Sara Nikseresht, Siena Barton, Khairul Ansari, Sukhdeep Spall, Laura F Dagley, Susan Belzer, Gregory Rice.
    High-throughput isolation and enrichment of extracellular vesicles using an immunoaffinity magnetic bead-based matrix. Presented at ISEV Conference in Seattle, USA (19 May 2023).
  5. Poster Presentation: Ramin Khanabdali, Chun Liu, Carlos Palma, Saso Ivanovski, Pingping Han, Gregory Rice. EXO-NET enriched salivary extracellular vesicles in periodontitis. Presented at ISEV Conference in Seattle, USA (20 May 2023).

 

Abstracts

  1. Carlos Salomon, Saumya Das, Uta Erdbrügger, Raghu Kalluri, Sai Kiang Lim, Jerrold M Olefsky, Gregory E Rice, Susmita Sahoo, W Andy Tao, Pieter Vader, Qun Wang, Alissa M Weaver.
    Extracellular Vesicles and Their Emerging Roles as Cellular Messengers in Endocrinology: An Endocrine Society Scientific Statement. Endocrine Reviews (June 2022).

 

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