Logo: to the web site of Uppsala University

uu.sePublikasjoner fra Uppsala universitet
Endre søk
RefereraExporteraLink to record
Permanent link

Direct link
Referera
Referensformat
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Annet format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annet språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf
Characterization of HexaSynO2-scFv8D3 antibody targeting α-synuclein oligomers in Parkinson’s disease
Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Farmaceutiska fakulteten, Institutionen för farmaci. (ProDDe - Protein Drug Design)
2022 (engelsk)Independent thesis Advanced level (degree of Master (Two Years)), 30 poäng / 45 hpOppgave
Abstract [en]

Aggregation and oligomerization of the alpha-synuclein (α-syn) protein, encoded by the SNCA gene, leads to the formation of Lewy bodies, one of the main risk factors of the progression of the Parkinson’s disease (PD). With a prevalence of around 100-200/100.000 people a year, PD is the second most common neurodegenerative disorder. Among the variety of symptoms, the main ones include tremor, bradykinesia and depression. Studies proved that soluble oligomers of α-syn, could be a valid target for specific recombinant antibodies which are able to cross the blood brain barrier (BBB) and trigger the immune response against these species of aggregates.This study focused on the purification and characterization of a newly designed hexavalent SynO2 (Hexa-SynO2-scFv8D3), specific for binding α-syn protein, and linked to single chain fragment variable (scFv) of the 8D3 antibody (scFv8D3), which is specific to bind to the transferrin receptor and thus be able to cross the BBB. Ultimately, the potential usage of Hexa-SynO2-scFv8D3 as a PET ligand in vivo should have been elucidated. The antibody was purified from human endothelial cells supernatant (HEK293) using an Äkta start system. Purity and quality were determined in vitro by performing SDS PAGE, Western Blot and Dynamic light scattering (DLS) analysis. Conformational stability was tested by thermal shift assay with Tycho NT.6. Finally, binding properties and functional stability were investigated by performing an indirect enzyme-linked immunosorbent assay (ELISA). Hexa-SynO2-scFv8D3 proved to have a good thermal stability overtime, however, its functional stability decreased with time. In addition, the antibody showed a tendency to aggregate, and the final concentration and yield were relatively low, reason why it could not be tested in vivo. Additional studies must be performed in order to increase the final concentration and yield. A new tested purification method could potentially improve the yield.

sted, utgiver, år, opplag, sider
2022. , s. 28
HSV kategori
Identifikatorer
URN: urn:nbn:se:uu:diva-477454OAI: oai:DiVA.org:uu-477454DiVA, id: diva2:1671280
Fag / kurs
Pharmacy
Utdanningsprogram
Master Programme in Drug Discovery and Development
Veileder
Examiner
Tilgjengelig fra: 2022-06-17 Laget: 2022-06-17 Sist oppdatert: 2022-06-17bibliografisk kontrollert

Open Access i DiVA

Fulltekst mangler i DiVA

Av organisasjonen

Søk utenfor DiVA

GoogleGoogle Scholar

urn-nbn

Altmetric

urn-nbn
Totalt: 147 treff
RefereraExporteraLink to record
Permanent link

Direct link
Referera
Referensformat
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Annet format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annet språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf