Department of Biotechnology
Molecular Biotechnology
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  Affinity-protein Based Blocking of
Cellular Proteins and their Delivery
Summary
Eriksson Karlström
Gräslund
Nygren
Samuelson
Ståhl

Principal Investigator (PI): Torbjörn Gräslund, PhD

Staff: Camilla Hofström (Graduate student), Ali Sheikholvaezin (Post-doc), Erik Vernet (Graduate student)

Affibody molecules

Affibody molecules are small and robust affinity proteins. For many purposes they can be used to replace antibodies and other conventional affinity reagents and function in a similar manner. In addition, their behavior in for some applications can be superior, where the biophysical properties of the conventional affinity reagents are unsatisfactory regarding, for example, folding, stability, production and size.
Our work aims at generating affibody reagents to desired antigens and to employ them in different applications. Our current work has dealt with:
Development of affibody reagents binding to the intracellular oncogenic transcription factor c-Jun.
Comparison of affibody molecules and antibodies as reagents in standard biochemical applications such as flow cytometry, immunofluorescence microscopy and protein “pull-down” experiments.
Engineering of affibody molecules specific for the oncogenic HER-2 receptor to become stationary in the secretory compartments of cells. This lead to hindering of receptor export and to a decrease in cell growth.

The future focus of the project is to further develop affibody-based reagents to study and specifically interfere with biological processes.

Other interests
Development of strategies for cellular transduction, including virus-based methods and strategies based on cell penetrating peptides.
Stable and specific knock-down of protein function using siRNAs delivered by virus-based vectors.
Artificial transcription factors.
Some current publications
Gräslund T., Li X., Magnenat L., Popkov M., Barbas CF. 3rd. Exploring strategies for the design of artificial transcription factors: targeting sites proximal to known regulatory regions for the induction of gamma-globin expression and the treatment of sickle cell disease. J Biol Chem. 280(5):3707-14 (2005)
Hedhammar M., Gräslund T., Hober S. (invited review). Protein Engineering Strategies for Selective Protein Purification. Chem. Eng. & Tech. 28(11): 1315-1325 (2005)
Hedhammar M., Alm T., Gräslund T., Hober S. Single-step recovery and solid-phase refolding of inclusion body proteins using a polycationic purification tag.
Biotech. Journal 1(2):187-196 (2006)
Lundberg E., Höidén-Guthenberg I., Larsson B., Uhlén M., Gräslund T. Site-specifically conjugated anti-HER2 Affibody molecules as one-step reagents for target expression analyses on cells and xenograft samples. J Immunol Methods. 319(1-2):53-63 (2007)
Gordley RM., Smith JD., Gräslund T., Barbas CF. Evolution of programmable zinc finger-recombinases with activity in human cells. J Mol Biol.367(3):802-13 (2007)
Lundberg E., Sundberg M., Gräslund T., Uhlén M. and Andersson-Svahn H. A novel solid phase method for fluorescent labeling of antibodies. J Immunol Methods:322(1-2):40-9 (2007)
Vernet E, Konrad A, Lundberg E, Nygren PA, Gräslund T. Affinity-based entrapment of the HER2 receptor in the endoplasmic reticulum using an affibody molecule. J Immunol Methods. 338(1-2):1-6. (2008)
Lundberg E, Brismar H, Gräslund T. Selection and characterization of Affibody ligands to the transcription factor c-Jun. Biotechnol Appl Biochem. (2008).
Last updated: 2009-01-20