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Process development for the control of solubility of Affibody® molecules
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Biology Education Centre.
2011 (English)Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

In this study the aim was to optimize the production of the Affibody fusion-protein Z03358-

ABD094-(S4G)3-IL2 with regard to the amount of soluble protein produced. However,

problems with reproducibility with this protein and the chosen expression system were

encountered. Therefore, expression of the His-tagged Affibody His6-(Z05477)2 was

evaluated using the same expression system as well as expression in another well

characterized expression system.

Both target proteins are of therapeutic interest. One of the proteins is an IL2 fusion

protein (Z03358-ABD094-(S4G)3-IL2) that bind the platelet-derived growth factor receptor β

(PDGFR-β). PDGF signaling is of interest in cancer treatment where, among other things, the

effects of PDGF on tumor angiogenesis is researched. The His6-(Z05477)2 protein has a

classified target but is developed as a therapeutic in the area of inflammation and autoimmune

disease. Both model proteins are known to be difficult to purify due to low solubility.

The two E. coli expression systems investigated and compared were BL21(DE3) and

Lemo21(DE3). The fusion protein Z03358-ABD094-(S4G)3-IL2 was produced in

BL21(DE3) in inclusion bodies with a yield of 4.95 g/l. An optimized process for the

expression of His6-(Z05477)2 using BL21(DE3) was developed with a yield of 6.6 g/l soluble

protein after expression at 30°C for 6 h.

Place, publisher, year, edition, pages
2011. , 23 p.
Keyword [en]
Protein expression, Platelet derived growth factor receptor-β (PDGFR- β), Expression systems, High cell density cultivations (HCDC)
National Category
Pharmaceutical Biotechnology
Identifiers
URN: urn:nbn:se:uu:diva-160250OAI: oai:DiVA.org:uu-160250DiVA: diva2:448938
Subject / course
Läkemedel
Educational program
Molecular Biotechnology Engineering Programme
Uppsok
Technology
Supervisors
Examiners
Available from: 2011-10-18 Created: 2011-10-18 Last updated: 2011-10-18Bibliographically approved

Open Access in DiVA