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Produktion av HER2-HER3 bispecifika Affibodymolekyler för cellstudier in vitro
KTH, School of Biotechnology (BIO).
2014 (Swedish)Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesisAlternative title
Production of HER2-HER3 bispecific Affibody molecules for in vitro cell studies (English)
Abstract [en]

Overexpressin of HER2 is seen in a number of cancer types and is characterized by aggressive disease with poor prognosis. HER2 targeted therapeutics, such as monoclonal antibodies, are today established for the treatment of HER2 positive cancer types but resistance to these therapeutics are common leading to relapse of disease. This could be overcome by creating bispecific affinity proteins with the ability to simultaneously bind two tumour-associated antigens. In HER2 positive cancer cells coexpression of HER3 is seen. Moreover, HER2 and HER3 form a very potent oncogenic unit with overactive signalling which results in increased proliferation and survival of cells. This receptor pair is therefore an interesting target for the development of bispecific affinity proteins. In this project, bispecific and bivalent Affibody mnolecules targeting this heterodimer have been produced. The molecules, which have high affinity for HER2 and HER3, are fused to an albuminbinding domain (ABD). This domain was used as a purification tag for affinity purification after protein production in E.coli and will also prolong the in vivo halflife of the constructs. Successful protein production and purification of the constructs were followed by in vitro cell studies. In these, the antiproliferative effects of the constructs on the pancreatic cancer cell line BxPc3, which coexpresses HER2 and HER3, were studied. The results from these studies indicate that the Affibody molecules have antiproliferative effects on the cancer cells. By this, we believe that the constructs are interesting for in vivo studies.

Place, publisher, year, edition, pages
2014.
Keyword [en]
Bispecific, Affibody molecules, HER2, HER3, Cancer, Half-life extention, ffinity proteins
National Category
Engineering and Technology
Identifiers
URN: urn:nbn:se:kth:diva-163669OAI: oai:DiVA.org:kth-163669DiVA: diva2:801712
Supervisors
Examiners
Available from: 2015-04-14 Created: 2015-04-10 Last updated: 2015-09-17Bibliographically approved

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CiteExportLink to record
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Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf