ArticleProtein engineering, design & selection : PEDS2015
Directed evolution of anti-HER2 DARPins by SNAP display reveals stability/function trade-offs in the selection process.
Article in Protein engineering, design & selection : PEDS, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed, 31 citations in OpenAlex.
- A method for the detection and enrichment of endogenous cereblon substrates.Cell chemical biology · 2025Article
- A method for the detection and enrichment of endogenous cereblon substrates.bioRxiv : the preprint server for biology · 2025Article
- Cell-Free Gene Expression: Methods and Applications.Chemical reviews · 2025Review
- Computational design of newly engineered DARPins as HER2 receptor inhibitors for breast cancer treatment.Research in pharmaceutical sciences · 2023Article
- Engineered fast-dissociating antibody fragments for multiplexed super-resolution microscopy.Cell reports methods · 2022Article
- The production of the first functional antibody mimetic in higher plants: the chloroplast makes the DARPin G3 for HER2 imaging in oncology.Biological research · 2022Article
- Co-optimization of therapeutic antibody affinity and specificity using machine learning models that generalize to novel mutational space.Nature communications · 2022Article
- The Effect of Mutations in the TPR and Ankyrin Families of Alpha Solenoid Repeat Proteins.Frontiers in bioinformatics · 2021Review
- In vitro evolution of antibody affinity via insertional scanning mutagenesis of an entire antibody variable region.Proceedings of the National Academy of Sciences of the United States of America · 2020Article
- Split & mix assembly of DNA libraries for ultrahigh throughput on-bead screening of functional proteins.Nucleic acids research · 2020Article
- Directed evolution methods for overcoming trade-offs between protein activity and stability.AIChE journal. American Institute of Chemical Engineers · 2020Article
- Development of a novel anti-HER2 scFv by ribosome display and in silico evaluation of its 3D structure and interaction with HER2, alone and after fusion to LAMP2B.Molecular biology reports · 2018Article
- Understanding and overcoming trade-offs between antibody affinity, specificity, stability and solubility.Biochemical engineering journal · 2018Article
- High-throughput screening of biomolecules using cell-free gene expression systems.Synthetic biology (Oxford, England) · 2018Review
- Arginine mutations in antibody complementarity-determining regions display context-dependent affinity/specificity trade-offs.The Journal of biological chemistry · 2017Article
- Efficient affinity maturation of antibody variable domains requires co-selection of compensatory mutations to maintain thermodynamic stability.Scientific reports · 2017Article
- A Shorter Route to Antibody Binders via Quantitative in vitro Bead-Display Screening and Consensus Analysis.Scientific reports · 2016Article
- Co-evolution of affinity and stability of grafted amyloid-motif domain antibodies.Protein engineering, design & selection : PEDS · 2015Article
- Review
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In vitro display technologies have proved to be powerful tools for obtaining high-affinity protein binders. We recently described SNAP display, an entirely in vitro DNA display system that uses the SNAP-tag to link protein with its encoding DNA in water-in-oil emulsions. Here, we apply SNAP display for the affinity maturation of a designed ankyrin repeat proteins (DARPin) that binds to the extracellular domain of HER2 previously isolated by ribosome display. After four SNAP display selection cycles, proteins that bound specifically to HER2 in vitro, with dissociation constants in the low- to sub-nanomolar range, were isolated. In vitro affinities of the panel of evolved DARPins directly correlated with the fluorescence intensities of evolved DARPins bound to HER2 on a breast cancer cell line. A stability trade-off is observed as the most improved DARPins have decreased thermostability, when compared with the parent DARPin used as a starting point for affinity maturation. Dissection of the framework mutations of the highest affinity variant, DARPin F1, shows that functionally destabilising and compensatory mutations accumulated throughout the four rounds of evolution.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.