The DOI matches its Crossref record; 51 of the 51 references it cites were found in a registry or an index.
High-affinity binders selected from designed ankyrin repeat protein libraries
Abstract
We report here the evolution of ankyrin repeat (AR) proteins in vitro for specific, high-affinity target binding. Using a consensus design strategy, we generated combinatorial libraries of AR proteins of varying repeat numbers with diversified binding surfaces. Libraries of two and three repeats, flanked by 'capping repeats,' were used in ribosome-display selections against maltose binding protein (MBP) and two eukaryotic kinases. We rapidly enriched target-specific binders with affinities in the low nanomolar range and determined the crystal structure of one of the selected AR proteins in complex with MBP at 2.3 A resolution. The interaction relies on the randomized positions of the designed AR protein and is comparable to natural, heterodimeric protein-protein interactions. Thus, our AR protein libraries are valuable sources for binding molecules and, because of the very favorable biophysical properties of the designed AR proteins, an attractive alternative to antibody libraries.
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“These POI-targeting polypeptides can either be single chain antibodies (e.g. variable domains from camelid or shark-derived heavy chain only antibodies, nanobodies), or a variety of synthetic polypeptide scaffolds that bind POIs such as fibronectin type III domain-based monobodies [110, 111], designed ankyrin repeats (DARPINs) [112, 113], cystatin-derived affimers [114] or Affibodies derived from”
From Advances in targeted degradation of endogenous proteins (2019) · cites this paper as [113]
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A Protein-Tagging System for Signal Amplification in Gene Expression and Fluorescence Imaging
Tanenbaum, M. E., Gilbert, L. A., Qi, L. S., Weissman, J. S., & Vale, R. D. (2014) · Cell · doi:10.1016/j.cell.2014.09.039 · Cited by 1,611
Analysis of aptamer discovery and technology
Dunn, M. R., Jimenez, R. M., & Chaput, J. C. (2017) · Nature Reviews Chemistry · doi:10.1038/s41570-017-0076 · Cited by 938
Engineering novel binding proteins from nonimmunoglobulin domains
Binz, H. K., Amstutz, P., & Plückthun, A. (2005) · Nature Biotechnology · doi:10.1038/nbt1127 · Cited by 676
Self-citationAnkyrin Repeat: A Unique Motif Mediating Protein−Protein Interactions
Li, J., Mahajan, A., & Tsai, M. (2006) · Biochemistry · doi:10.1021/bi062188q · Cited by 651
Designed Ankyrin Repeat Proteins (DARPins): Binding Proteins for Research, Diagnostics, and Therapy
Plückthun, A. (2011) · The Annual Review of Pharmacology and Toxicology · doi:10.1146/annurev-pharmtox-010611-134654 · Cited by 572
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Metadata
- Authors
- Binz, H. K., Amstutz, P., Kohl, A., Stumpp, M. T., Briand, C., Forrer, P., Grütter, M. G., & Plückthun, A.
- Affiliations
- University of Zurich (CH)
- Corresponding author
- Grütter, Markus G
- Plückthun, Andreas
- Venue
- Nature Biotechnology
- Published
- 2004-04-18
- Volume
- 22
- Issue
- 5
- Pages
- 575-582
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- OpenAlex
- W2084938664
- Cited by
- 656
- References
- 51
- Citations by year
- 2026: 7 · 2025: 22 · 2024: 17 · 2023: 26 · 2022: 27 · 2021: 23 · 2020: 31 · 2019: 24 · 2018: 22 · 2017: 38 · 2016: 28 · 2015: 36 · 2014: 33 · 2013: 33 · 2012: 35
- FWCI
- 30.73
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Bundesministerium für Bildung und Forschung
- National Center of Competence in Research Chemical Biology
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › Monoclonal and Polyclonal Antibodies Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Structural Characterization
- Physical Sciences › Chemistry › Organic Chemistry › Chemical Synthesis and Analysis
- Keywords
- ankyrin repeat proteins, combinatorial protein libraries, ribosome display, protein-protein interaction, maltose-binding protein, high-affinity binders, consensus design, protein crystal structure
- MeSH terms
- Amino Acid Sequence, Enzyme-Linked Immunosorbent Assay, Molecular Sequence Data, Protein Conformation, Ankyrin Repeat
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