Protein corona formation and structural changes of lipid nanoparticles investigated using QCM-D and waveguide microscopy
| dc.contributor.author | Mohammadi, Sara | |
| dc.contributor.department | Chalmers tekniska högskola / Institutionen för fysik | sv |
| dc.contributor.examiner | Höök, Fredrik | |
| dc.contributor.supervisor | Höök, Fredrik | |
| dc.contributor.supervisor | Sjöberg, Mattias | |
| dc.contributor.supervisor | Agnarsson, Björn | |
| dc.date.accessioned | 2021-06-08T15:12:16Z | |
| dc.date.available | 2021-06-08T15:12:16Z | |
| dc.date.issued | 2021 | sv |
| dc.date.submitted | 2020 | |
| dc.description.abstract | Lipid nanoparticles (LNPs) are being increasingly considered for use in drug delivery. However, understanding the interactions of LNPs with a biological environment is crucial for successful deliver of their cargo, such as for example mRNA, across cellular membranes. Upon exposure to a biological fluid it is believed that the surface of LNPs is spontaneously covered by a protein corona, a layer of adhered biomolecules, prior to being taken up by the targeted cells. A protein of interest in this context is Apolipoprotein-E (ApoE), both because it is known to be part of the protein corona and because it may control endocytic uptake via specific interactions with receptors in the cell membrane. Furthermore, ApoE-coated LNPs are expected to undergo a structural change in the acidic environment of the endosome during cargo release to the cytosol. In this master thesis, we have studied protein-corona formation and the reaction of the LNPs covered/not covered with ApoE in an acid environment that mimics the endosomal environment using label-free surface-based bioanalytical tools such as quartz crystal microbalance with dissipation monitoring (QCM-D) and waveguide microscopy. QCM-D provides information based on ensemble averaging of the biological interactions taking place on the surface of the sensor, while waveguide microscopy provides both ensemble-averaged data and information based on inspection of individual LNPs. Further, waveguide microscopy is capable of detecting both light scattering and fluorescence signals, which provides the possibility to observe both the labeled interior of the LNPs and binding of unlabeled protein. We found that ApoE bind to PEG-modified LNPs with a lag-time of tens of minutes unless mixed with bovine serum albumin (BSA). This is attributed to BSA-induced PEG shedding which promotes ApoE binding. Inspections using combined labelfree and fluorescence based waveguide microscopy releveled a weaker than expected dependence between cargo fluorescence and scattering intensity, suggesting that the self-assembly process utilized for LNP fabrication may vary with LNP size. It is also concluded that further liquid handling must be improved to follow ApoE binding to LNPs using waveguide microscopy. | sv |
| dc.identifier.coursecode | TIFX05 | sv |
| dc.identifier.uri | https://hdl.handle.net/20.500.12380/302413 | |
| dc.language.iso | eng | sv |
| dc.setspec.uppsok | PhysicsChemistryMaths | |
| dc.subject | Waveguide microscopy | sv |
| dc.subject | Quartz crystal microbalance with dissipation monitoring | sv |
| dc.subject | label-free | sv |
| dc.subject | apolipoprotein-E | sv |
| dc.subject | corona formation | sv |
| dc.subject | protein corona | sv |
| dc.subject | lipid nanoparticle | sv |
| dc.subject | MC3 | sv |
| dc.subject | drug delivery | sv |
| dc.subject | mRNA therapy | sv |
| dc.subject | serum protein | sv |
| dc.subject | endosomal membrane | sv |
| dc.title | Protein corona formation and structural changes of lipid nanoparticles investigated using QCM-D and waveguide microscopy | sv |
| dc.type.degree | Examensarbete för masterexamen | sv |
| dc.type.uppsok | H |