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FRET-Based Assay fo...
FRET-Based Assay for the Quantification of Extracellular Vesicles and Other Vesicles of Complex Composition
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- Thorsteinsson, Konrad, 1991- (författare)
- Umeå universitet,Avdelningen för virologi,Wallenberg centrum för molekylär medicin vid Umeå universitet (WCMM),Marta Bally,Umeå University
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- Olsén, Erik, 1994 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology,Division of Nano and Biophysics, Department of Physics, Chalmers University of Technology, Gothenburg, Sweden
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- Schmidt, Eneas, 1991 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology,Division of Nano and Biophysics, Department of Physics, Chalmers University of Technology, Gothenburg, Sweden
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- Pace, Hudson, 1982 (författare)
- Umeå universitet,Wallenberg centrum för molekylär medicin vid Umeå universitet (WCMM),Avdelningen för virologi,Umeå University
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- Bally, Marta, 1981 (författare)
- Umeå universitet,Wallenberg centrum för molekylär medicin vid Umeå universitet (WCMM),Avdelningen för virologi,Marta Bally,Umeå University
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(creator_code:org_t)
- 2020-11-12
- 2020
- Engelska.
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Ingår i: Analytical Chemistry. - : American Chemical Society (ACS). - 0003-2700 .- 1520-6882. ; 92:23, s. 15336-15343
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Abstract
Ämnesord
Stäng
- Research in the field of extracellular vesicles is rapidly expanding and finding footholds in many areas of medical science. However, the availability of methodologies to quantify the concentration of membrane material present in a sample remains limited. Herein, we present a novel approach for the quantification of vesicle material, specifically the quantification of the total lipid membrane surface area, found in a sample using Förster resonance energy transfer (FRET). In this assay, sonication is used to drive the fusion between vesicles in the sample to be quantified and liposomes containing a pair of FRET fluorophores. The change in emission spectrum upon vesicle fusion is directly related to the total membrane surface area of the sample added, and a calibration curve allows for the quantification of a variety of vesicle species, including enveloped viruses, bacterial outer membrane vesicles, and mammalian extracellular vesicles. Without extensive optimization of experimental parameters, we were able to quantify down to ∼109 vesicles/mL, using as little as 60 μL of the sample. The assay precision was comparable to that of a commercial nanoparticle tracking analysis system. While its limit of detection was slightly higher, the FRET assay is superior for the detection of small vesicles, as its performance is vesicle-size-independent. Taken together, the FRET assay is a simple, robust, and versatile method for the quantification of a variety of purified vesicle samples.
Ämnesord
- NATURVETENSKAP -- Kemi -- Fysikalisk kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences -- Physical Chemistry (hsv//eng)
- NATURVETENSKAP -- Kemi -- Analytisk kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences -- Analytical Chemistry (hsv//eng)
- TEKNIK OCH TEKNOLOGIER -- Elektroteknik och elektronik -- Annan elektroteknik och elektronik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Electrical Engineering, Electronic Engineering, Information Engineering -- Other Electrical Engineering, Electronic Engineering, Information Engineering (hsv//eng)
Publikations- och innehållstyp
- art (ämneskategori)
- ref (ämneskategori)
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