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Principal Investigator (PI): Prof.
Per-Åke Nygren
Personnel: Sebastian Grimm (PhD student),
Feifan Yu (PhD student)
Description: The research
group has a present focus on the use of combinatorial protein
engineering principles to provide new tools for a variety
of biotechnology applications, with an emphasis on affinity
biotechnology. A core technology is an in-house developed
platform for the generation of tailor-made affinity proteins,
denoted affibodies. These are isolated by different powerful
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, in vivo diagnostics (imaging) and therapy.
The group is also exploring different means for construction
and handling of large gene/protein libraries during selections,
including phage display, ribosome display and protein fragment
complementation assay (PCA).
| Some key
publications: |
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Nord, K., Nilsson, J., Nilsson, B., Uhlén,
M and Nygren, P.-Å. (1995) "A
combinatorial library of an α-helical bacterial receptor
domain" Protein Engineering 8, 601-608. |
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Makrides, S., Nygren,
P.-Å., Andrews, B., Ford, P., Evans, K.S.,
Hayman, E.G., Adari, H., Levin, J., Uhlén, M. and
Toth, C.A. (1996) "Extended in vivo half-life
of human soluble complement receptor type I fused to serum
albumin-binding receptor" J. Pharm. Exp. Therap.
277, 534-541. |
| • |
Jendeberg, L., Nilsson, P., Larsson, A.,
Nilsson, B., Uhlén, M. and Nygren, P.-Å.
(1997) "Engineering of Fc1 and Fc3 from human immunoglobulin
G to analyse subclass specificity for staphylococcal protein
A" J. Imm. Methods 201, 25-34. |
| • |
Nord, K., Gunneriusson, E., Ringdahl, J.,
Ståhl, S., Uhlén, M. and Nygren, P.-Å.
(1997) "Binding proteins selected from libraries of
an α-helical bacterial receptor domain" Nature
Biotechnology 15, 772-777. |
| • |
Jansson, B., Uhlén, M. and Nygren,
P.-Å. (1998) "All individual domains
of staphylococcal protein A show Fab binding" FEMS
Immunol. Med. Microbiol. 20, 69-78. |
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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. |
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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. |
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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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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. |
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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 |
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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. |
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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. |
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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.
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Johanson, M. U., Frick, I.-M., Nilsson, H.,
Kraulis, P. J., Hober, S., Jonasson, P., Nygren,
P.-Å., Uhlén, M., Björck, L.,
Drakenberg, T., Forsén, S. and Wikström, M.
"Structure, specificity and mode of interaction for
bacterial albumin-binding modules" J. Biol. Chem.
277, 8114-8120. |
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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. |
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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. |
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Eklund, M., Axelsson, L., Uhlén, M
and Nygren, P.-Å. (2002) "Anti-idiotypic
protein domains selected from Protein A-based affibody libraries"
Prot. Struct. Funct. Gen. 48, 454-462. |
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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. |
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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. |
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Gräslund, S., Eklund, M., Falk, R.,
Uhlén, M., Nygren, P.-Å. and
Ståhl, S. (2002) "A novel affinity gene system
allowing protein A-based recovery of non-immunoglobulin
gene products" J. Biotechnol. 99, 41-50. |
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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. |
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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. |
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| Rönnmark, J, Kampf, C., Asplund, A.,
Höidén-Guthenberg, Wester, K., I-M, Pontén,
F., Uhlén, M. and Nygren, P.-Å.
(2003)"Affibody-β-galactosidase immunoconjugates
produced as soluble fusion proteins in the E. coli
cytosol". J. Immunol. Meth. 281, 149-160. |
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| 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. |
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| Nord, O., Uhlén, M. and Nygren,
P.-Å. (2003) "Microbead display of proteins
by cell-free expression of anchored DNA" J. Biotechnology
106, 1-13. |
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| Andersson, M., Rönnmark, J., Nygren,
P.-Å. and Ahlborg, N. (2003) "Inclusion
of a non-antibody binding protein in two-site ELISAs for
quantification of human serum proteins without interference
by heterophilic serum antibodies" J. Immunol. Meth.
283, 225-234. |
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| Eklund, M., Sandström K., Teeri, T.
and Nygren, P.-Å. (2004) "Site-specific
and reversible anchoring of active proteins onto cellulose
using a cellulosome-like complex" J. Biotechnol.
109, 277-286. |
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| Renberg, B., Nygren, P.-Å.,
Eklund, M. and Eriksson Karlström, A. (2004) "Fluorescence
resonance energy transfer-based detection of analytes using
anti-idiotypic affinity protein pairs" Anal. Biochem.
334, 72-80. |
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| Nygren, P.-Å and Skerra
A. (2004) "Binding proteins from alternative scaffolds"
J. Immunol. Meth. 290, 3-28. |
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| Nord, O., Gustrin, A. and Nygren,
P.-Å. (2005) "Fluorescent detection
of β-lactamase activity in living E. coli
via esterase supplementation" FEMS Microbiology
Letters 242, 73-79. |
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| Engfeldt, T., Renberg B., Brumer, H., Nygren,
P.-Å. and Eriksson Karlström, A. "Chemical
synthesis of fluorescent-labeled affinity proteins for specific
binding and detection of unlabeled protein" Chem.
BioChem. 6, 1043-1050. |
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| Renberg, B., Shiroyama, I., Engfeldt, T.,
Nygren, P.-Å. and Eriksson Karlström,
A. (2005) "Affibody protein capture microarrays: synthesis
and evaluation of random and directed immobilization of
affibody molecules" Anal. Biochem. 334, 72-80. |
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| Renberg, B., Nordin, J. Merca, A., Uhlén,
M., Feldwisch, J., Nygren, P.-Å.
and Eriksson Karlström, A. (2007) "Affibody molecules
in protein capture microarrays. evaluation of multidomain
ligands and different detection formats" J. Proteome
Res. 6, 171-179. |
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| Taussig MJ, Stoevesandt O, Borrebaeck CA,
Bradbury AR, Cahill D, Cambillau C, de Daruvar A, Dubel
S, Eichler J, Frank R, Gibson TJ, Gloriam D, Gold L, Herberg
FW, Hermjakob H, Hoheisel JD, Joos TO, Kallioniemi O, Koegl
M, Konthur Z, Korn B, Kremmer E, Krobitsch S, Landegren
U, van der Maarel S, McCafferty J, Muyldermans S, Nygren
P-Å, Palcy S, Pluckthun A, Polic B, Przybylski
M, Saviranta P, Sawyer A, Sherman DJ, Skerra A, Templin
M, Ueffing M, Uhlen M. (2007) "ProteomeBinders: planning
a European resource of affinity reagents for analysis of
the human proteome" Nat Methods 4, 13-17. |
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| Bandmann N. and Nygren P-Å.
(2007) "Combinatorial expression vector engineering
for tuning of recombinant protein production in Escherichia
coli" Nucleic Acids Res. 35(5):e32. |
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| Jonsson, A., Dogan J., Herne, N., Abrahmsen,
L. and Nygren, P.-Å. (2008) "Engineering
of a femtomolar affinity binding protein to human serum
albumin" Protein Engineering, Design and Selection
(PEDS) 275, 2668-2676. |
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| Nygren, P.-Å. (2008)
"Alternative binding proteins: Affibody binding proteins
developed from a small three-helix bundle scaffold"
FEBS Journal 275, 2668-2676. |
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| Vernet, E., Konrad, A., Lundberg, E, Nygren,
P.-Å. and Gräslund, T. "Affinity-based
entrapment of the HER2 receptor in the endoplasmatic reticulum
using an affibody molecule" Submitted. |
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