Characterization of his-mumps nucleocapsid protein virus-like particles
Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesisAlternative title
Karaktärisering av vinslika partiklar baserat på nukleokapsidprotein från påssjukevirus (Swedish)
A protocol for production of his-mumps nucleocapsid virus-like parcticles has together with an external collaborating company, been created by Andres Veide and Johan Norén (KTH)- The purification process has suffered from unexpected yield losses with the hypothesis that tha virus-like particles shred on a final downstream processing filtration step. The aim of this thesis, apart from to verify or discard the hypothesis, has been to characterize the mumps nucleocapsid protein with respect to its size and self-assembly.
The particles were characterized using dynamic light scattering, optical density, size exclusion chromatography and gel electrophoresis. The results indicates that the virus-like particles clute from the primary purification step, IMAC, as a donut-shpaed structure and due to the following buffer exchange to 50 mM HEPES assembles into a cylinder-shaped structure. It is thus not the filtration in itself that affects the protein; it is rather the buffer exchange. This thesis suggests a way of manipulating the size of the particles by changing buffer composition. Further work must sill be done, and analysis with transmission electron microscopy is of certain interest.
Place, publisher, year, edition, pages
Mumps, virus-like particle, nucleocapsid protein, dynamic light scattering, protein interaction
Engineering and Technology
IdentifiersURN: urn:nbn:se:kth:diva-150486OAI: oai:DiVA.org:kth-150486DiVA: diva2:744719