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Micro-Nanofluidic Isolation of Circulating Exosomes from Blood Plasma

Abdorahimzadeh, Amir Hossein | 2020

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  1. Type of Document: M.Sc. Thesis
  2. Language: Farsi
  3. Document No: 52814 (08)
  4. University: Sharif University of Technology
  5. Department: Mechanical Engineering
  6. Advisor(s): Taghipoor, Mojtaba; saeedi, Mohammad saeed
  7. Abstract:
  8. Exosomes are a type of extracellular vesicles which are secreted by cells and could be found in all of human biofluids including blood. The size of exosomes is in the range of 30 – 150 nm. And they have a spherical structure with a phospholipid membrane. Exosomes are very important in academic research for the purpose of diagnostic and therapeutic practices because of their protein and nucleic acid contents.In this study a technological system was developed for isolation of circulating exosomes of human whole blood. This system consisted of a collection of microfluidic chips and protocols in order to extract a purified sample of exosomes from whole blood. First, a chip based on centrifugal platform was introduced to separate and filter blood plasma. Using centrifugal force, this chip first separated plasma from red blood cells; then, it discarded particles larger than 220 nm from the separated plasma using a filter with 220 nm pore size. In the next stage, a microfluidic chip based on the pressure driven platform was designed and fabricated. The function of this chip was based on tangential flow filtration of proteins in the plasma sample filtered in the previous stage. Using a filter with 30 nm pore size, this chip removed protein contaminations in the filtered plasma sample and purified the remaining exosomes. In order to evaluate the functioning of this system, the final samples extracted were examined with the help of DLS, SDS-PAGE and Western blot tests. Primary results indicated that the system is capable of eliminating excessive proteins in the exosome sample and preserving exosomes in the sample through the process of filtration. Regarding low expenses in making the chips and the simplicity of working with the proposed protocols, this system has the potentiality of being exploited instead of commonly used exosome isolation methods including ultracentrifugation and size exclusion chromatography which are known as being expensive and complicated
  9. Keywords:
  10. Microfluidic Chip ; Cross Flow Microfiltration ; Lab-on-a-CD (LOCD) ; Dynamic Light Scattering ; Extracellular Vesicles ; Exosomes

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