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Design och karaktärisering av biparatopa Affibodymolekyler mot VEGFR2
KTH, School of Biotechnology (BIO).
2015 (Swedish)Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesisAlternative title
Engineering and characterization of biparatopic VEGFR2-specific Affibody molecules (English)
Abstract [en]

The physiological process of new blood vessel development from pre-existing ones is referred to as angiogenesis, and it is one of the most important contributors to cancer cell survival, proliferation and metastasis. Anti-angiogenesis therapeutic strategies have therefore become popular and possess considerable potential for treating angiogenesis related diseases such as cancer and ophtalmic disorders. Biparatopic Affibody constructs which target the Vascular Endothelial Growth Receptor-2 (VEGFR2), an important regulator of angiogenesis, have previously been generated with the intent of therapeutic and imaging utility. The small size of the Affibody provides several advantages over larger proteins, such as faster tissues penetration, higher stability and possible use of alternative administrative routes, which in this case could be the use of eye drops instead of intravitreal eye injections. Here we study how varying the linker length between the two domains in the biparatopic Affibody construct affects the inhibitory ability and select a candidate to move forward with for further anti-angiogenic studies. Both human and murine VEGFR2 binding was observed for all Affibody constructs. The construct chosen for further studies demonstrated inhibition of VEGFR2 phosphorylation, angiogenesis sprout formation of HUVECs and also cell proliferation of 293/KDR cells. The results indicate that the biparatopic Affibody holds potential for in vivo therapeutic and imaging utilization. New constructs of the Affibody were therefore designed, produced and conjugated to chelators and fluorphores for future studies.

Place, publisher, year, edition, pages
Keyword [en]
Angiogenesis, VEGFR2, Affibody molecule, Cancer, Ocular disorders
National Category
Engineering and Technology
URN: urn:nbn:se:kth:diva-173214OAI: diva2:851832
Available from: 2015-09-07 Created: 2015-09-07 Last updated: 2015-09-07Bibliographically approved

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