Research group: Combinatorial
protein engineering
Nord, K., Gunneriusson, E., Uhlén, M. and Nygren, P.-Å. (2000) "Ligands
selected from combinatorial libraries of protein a for use in affinity
capture of apolipoprotein A-1 and Taq DNA polymerase" J.
Biotechnol. 80, 45-54.
Lehtiö, J., Teeri, T. and Nygren, P.-Å. (2000) "Alpha-amylase inhibitors
selected from a combinatorial library of a cellulose binding domain
scaffold" Proteins: Struct. Funct. Gen. 41, 316-322
Principal Investigator (PI ): Prof. PerÅke Nygren
Danielsen S., Eklund, M., Deussen, H.-J., Gräslund, T., Nygren, P.-Å. and
Borchert, T. [2001] "In vitro selection of enzymatically active lipase
variants from phage libraries using a mechanism-based inhibitor"
Gene 272, 267-274.
Personnel: Malin Eklund, Olof Nord,
Erik Forsén, Nina Bandmann
Karlström, A. and Nygren, P.-Å. [2001] "Dual labeling of a binding protein
allows for specific fluorescence detection of native protein" J. Anal.
Biochem. 295, 22-30.
Description: Our research group has a
focus on the use and development of
combinatorial protein engineering
principles to provide new tools for a
variety of biotechnology applications,
with an emphasis on affinity
technology,
including
protein
expression, purification, detection and
biomolecular recognition. A core
technology is an in-house developed
platform for the generation of tailormade affinity proteins, denoted
affibodies. These are isolated by
different powerful in-house-developed
in vitro or in vivo selection principles
from large collections (libraries)
constructed
by
combinatorial
mutagenesis of a small and robust
protein domain used as scaffold. Such
"artificial antibodies" are at present
explored for use in a variety of
applications,
incl.
bioseparation,
diagnostics, self-assembly and therapy.
Combinatorial principles are also
employed for gene expression cassette
design to facilitate high-throughput
production of recombinant proteins.
Nord, K., Nord, O., Uhlén, M., Kelley, B., Ljungqvist, C. and Nygren, P.Å. (2001) “Recombinant human factor VIII-specific affinity ligands
selected from phage-displayed combinatorial libraries of protein A"
Eur. J. Biochem. 268, 4269-4277.
Bandmann, N., Van Alstine, J., Veide, A. and Nygren, P.-Å. (2002)
"Functional selection of phage displayed peptides for improved
aqueous two phase partitioning of recombinant proteins" J.
Biotechnology 93, 1-14.
Rönnmark, J., Hansson, M., Nguyen, T., Uhlén, M., Robert, A., Ståhl, S.
and Nygren, P.-Å. (2002) "Construction and characterization of
affibody-Fc chimeras produced in Escherichia coli" J. Immunol. Meth.
261, 199-211.
M. Eklund, Axelsson, L., Uhlén, M and Nygren, P.-Å. (2002) "Antiidiotypic protein domains selected from Protein A-based affibody
libraries" Prot. Struct. Funct. Gen.48, 454-462.
Rönnmark, J., Grönlund, H. Uhlén, M. and Nygren, P.-Å. (2002) "Human
immunoglobulin A (IgA)-specific ligands from combinatorial
engineering of protein A" Eur. J. Biochem. 269, 2647-2655.
Henning, P., Magnusson, M. K., Gunneriusson, E., Hong, S. S., Boulanger,
P., Nygren, P.-Å. and Lindholm, L. (2002) "Genetic modification of
adenovirus 5 tropism by a novel class of ligands based on a three-helix
bundle scaffold derived from staphylococcal protein A." Human Gene
Therapy 13, 1427-1439.
Rönnmark, J, Kampf, C., Asplund, A., Höidén-Guthenberg, Wester, K., IM, Pontén, F., Uhlén, M. and Nygren, P.-Å. (2003)"Affibody-galactosidase immunoconjugates produced as soluble fusion proteins
in the E. coli cytosol". J. Immunol. Meth. 281, 149-160.
Högbom, M., Eklund, M., Nygren, P.-Å. and Nordlund P. (2003) "Crystal
structure of an in vitro evolved affibody-protein complex" Proc. Natl.
Acad. Sci. U.S.A. 100, 3191-3196.
Wahlberg, E., Lendel, C., Helgstrand, M., Allard, P., Dincbas-Renqvist, V.,
Hedqvist, A., Berglund, H., Nygren, P.-Å. and Härd, T. (2003) "An
affibody in complex with a target protein: structure and coupled
folding" Proc. Natl. Acad. Sci. U.S.A. 100, 3185-3190.
Nord, N., Uhlén, M. and Nygren, P.-Å. (2003) "Microbead display of
proteins by cell-free expression of anchored DNA" J. Biotechnology
106, 1-13.
Sandström, K., Xu, Z., Forsberg, G. and Nygren, P.-Å. (2003) "Inhibition
of the CD28-CD80 co-stimulation signal by a CD28-binding ligand
developed by combinatorial protein engineering". Protein Engineering
6, 691-697.
Nord, K., Gunneriusson, E., Ringdahl, J., Ståhl, S., Uhlén, M. and
Nygren, P.-Å. (1997) "Binding proteins selected from libraries of
an -helical bacterial receptor domain" Nature Biotechnology 15,
772-777.
Eklund, M., Sandström K., Teeri, T. and Nygren, P.-Å. (2004) "Sitespecific and reversible anchoring of active proteins onto cellulose
using a cellulosome-like complex" J. Biotechnol. In press.
Nygren, P.-Å. and Uhlén, M. (1997) "Scaffolds for engineering novel
binding sites in proteins" Curr. Opin. Struct. Biol. 7, 463-469.
Hansson, M., Ringdahl, J., Robert, A., Power, U., Goetsch, L., Nguyen, T.,
Uhlén, M., Ståhl, S. and Nygren, P.-Å. (1999) "An in vitro selected
binding protein (affibody) shows conformation-dependent recognition
of the respiratory syncytial virus (RSV) G protein. Immunotechnology
4, 237-252.
Gunneriusson, E., Nord, K., Uhlén, M. and Nygren, P.-Å. (1999) "Affinity
maturation of a Taq DNA polymerase specific affibody by helix
shuffling" Protein Engineering 12, 873-878.
Legendre, D., Laraki, N., Gräslund, T., Björnvad, M. E., Bouchet, M.,
Nygren, P.-Å., Borchert, T. V. and Fastrez, J. (2000) “Display of
active subtilisin 309 on phage: Analysis of parameters influencing the
selection of subtilisin variants with changed substrate specificity from
libraries using phosphonylating inhibitors” J. Mol. Biol. 296, 87-102.
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