University of Twente Student Theses
Isolation of HepG2-derived extracellular vesicles in spiked human blood serum using magnetic beads and click chemistry
Geraets, E.K.W. (2023) Isolation of HepG2-derived extracellular vesicles in spiked human blood serum using magnetic beads and click chemistry.
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| Abstract: | Hepatocellular carcinoma (HCC) is a top contributor to cancer-related deaths making early diagnosis crucial. However, current surveillance techniques are inadequate in early stages of HCC. Extracellular vesicles (EVs) are lipid bilayers and potential biomarkers. Using current methods, it remains difficult to isolate EVs from blood samples. The aim of this research is to test immunomagnetic isolation of HepG2-derived EVs from spiked serum samples using click chemistry, magnetic beads and antibodies. This is achieved through a tetrazine (Tz) and trans-cyclooctene (TCO) reaction. EpCAM antibodies were successfully conjugated to magnetic beads which led to an estimation of 19.9% of beads positive for EVs, during isolation from culture medium. Thereafter, the system was tested in serum samples spiked with HepG2-derived EVs. Bead populations positive for EVs were estimated to be 21.3%, 9.5% and 5.4% for samples of EV-spiked PBS, 50% serum in PBS and 100% serum. In addition, free thiol groups on the magnetic beads were quantified using an Ellman's assay. To conclude, contaminants in serum decrease the capture efficiency of immunomagnetic beads while capturing HepG2-derived EVs. After increasing efficiency, the immunomagnetic isolation of EVs from serum samples is a promising, quick and low-risk surveillance method for early-stage diagnosis of HCC in patients. |
| Item Type: | Essay (Bachelor) |
| Faculty: | TNW: Science and Technology |
| Subject: | 35 chemistry, 42 biology, 44 medicine, 50 technical science in general |
| Programme: | Biomedical Technology BSc (56226) |
| Link to this item: | https://purl.utwente.nl/essays/96254 |
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