University of Twente Student Theses
Isolation of Hepatocellular carcinoma (HCC) derived extracellular vesicles Using click-chemistry conjugated immunomagnetic beads
Mentink, Sven (2023) Isolation of Hepatocellular carcinoma (HCC) derived extracellular vesicles Using click-chemistry conjugated immunomagnetic beads.
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| Abstract: | Hepatocellular carcinoma (HCC) is one of the top five most common malignant tumour types in the western world. The asymptomatic nature of early stage HCC leads to an issue that most patients are diagnosed at late stages of the disease, in which treatments are far less effective. Tumour derived extracellular vesicles (EVs) have been identified as novel liquid biopsy markers for early diagnosis and prognosis in HCC. However, current methods are insufficient for specific isolation of these HCC derived EVs. Here, we aim to develop a system that is capable of specifically capturing HCC derived EVs from conditioned cell medium. By combining different EV characterisation techniques initial presence of HCC derived EVs in conditioned cell medium could be confirmed. The developed capturing system is based on magnetic beads covalently conjugated to antibodies by Tetrazine (Tz) trans-cyclooctyne (TCO) mediated “click” reaction. Fluorescence activated cell sorting (FACS) and microscopy confirmed successful conjugation of subparts of the system, by making use of the inherent fluorescent properties of Tz. Subsequently, by successful staining of EVs with Calcein, FACS showed the capturing of HCC derived EVs from HepG2 cell conditioned medium. |
| Item Type: | Essay (Master) |
| Faculty: | TNW: Science and Technology |
| Subject: | 35 chemistry, 42 biology, 44 medicine |
| Programme: | Biomedical Engineering MSc (66226) |
| Link to this item: | https://purl.utwente.nl/essays/95808 |
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