ArticlePloS one2012
Design of pH sensitive binding proteins from the hyperthermophilic Sso7d scaffold.
Article in PloS one, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 33 papers.
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Who cites it
33 citing papers in PubMed, 56 citations in OpenAlex.
- Chemical modifications to enhance the stability of DNA nanoparticles for biological applications.iScience · 2026Review
- Identification and characterization of vasoactive intestinal peptide receptor antagonists with high-affinity and potent anti-leukemia activity.The Journal of biological chemistry · 2026Article
- Engineering and Structural Elucidation of a Sac7d-Derived IgG Fc-Specific Affitin and Its Application for the Light-Controlled Affinity Purification of Antibodies.Chembiochem : a European journal of chemical biology · 2025Article
- Isolation of nucleic acids using liquid-liquid phase separation of pH-sensitive elastin-like polypeptides.Scientific reports · 2024Article
- Strategies for Conditional Regulation of Proteins.JACS Au · 2023Review
- Engineering Strategies to Overcome the Stability-Function Trade-Off in Proteins.ACS synthetic biology · 2022Review
- Yeast Display Guided Selection of pH-Dependent Binders.Methods in molecular biology (Clifton, N.J.) · 2022Article
- A stepwise mutagenesis approach using histidine and acidic amino acid to engineer highly pH-dependent protein switches.3 Biotech · 2022Article
- Driving CARs with alternative navigation tools - the potential of engineered binding scaffolds.The FEBS journal · 2021Review
- Genetically encoded live-cell sensor for tyrosinated microtubules.The Journal of cell biology · 2020Article
- Article
- A Generic Procedure for the Isolation of pH- and Magnesium-Responsive Chicken scFvs for Downstream Purification of Human Antibodies.Frontiers in bioengineering and biotechnology · 2020Article
- Design and evaluation of engineered protein biosensors for live-cell imaging of EGFR phosphorylation.Science signaling · 2019Article
- De novo design of tunable, pH-driven conformational changes.Science (New York, N.Y.) · 2019Article
- Engineering Allostery into Proteins.Advances in experimental medicine and biology · 2019Review
- Engineering a Protein Binder Specific for p38α with Interface Expansion.Biochemistry · 2018Article
- Beyond antibody engineering: directed evolution of alternative binding scaffolds and enzymes using yeast surface display.Microbial cell factories · 2018Review
- Inefficient Ribosomal Skipping Enables Simultaneous Secretion and Display of Proteins in Saccharomyces cerevisiae.ACS synthetic biology · 2017Article
- Derivative of Extremophilic 50S Ribosomal Protein L35Ae as an Alternative Protein Scaffold.PloS one · 2017Article
- Fc-Binding Ligands of Immunoglobulin G: An Overview of High Affinity Proteins and Peptides.Materials (Basel, Switzerland) · 2016Review
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We have engineered pH sensitive binding proteins for the Fc portion of human immunoglobulin G (hIgG) (hFc) using two different strategies - histidine scanning and random mutagenesis. We obtained an hFc-binding protein, Sso7d-hFc, through mutagenesis of the Sso7d protein from the hyperthermophilic archaeon Sulfolobus solfataricus; Sso7d-hFc was isolated from a combinatorial library of Sso7d mutants using yeast surface display. Subsequently, we identified a pH sensitive mutant, Sso7d-his-hFc, through systematic evaluation of Sso7d-hFc mutants containing single histidine substitutions. In parallel, we also developed a yeast display screening strategy to isolate a different pH sensitive hFc binder, Sso7d-ev-hFc, from a library of mutants obtained by random mutagenesis of a pool of hFc binders. In contrast to Sso7d-hFc, both Sso7d-his-hFc and Sso7d-ev-hFc have a higher binding affinity for hFc at pH 7.4 than at pH 4.5. The Sso7d-mutant hFc binders can be recombinantly expressed at high yield in E. coli and are monomeric in solution. They bind an epitope in the CH3 domain of hFc that has high sequence homology in all four hIgG isotypes (hIgG(1-4)), and recognize hIgG(1-4) as well as deglycosylated hIgG in western blotting assays. pH sensitive hFc binders are attractive candidates for use in chromatography, to achieve elution of IgG under milder pH conditions. However, the surface density of immobilized hFc binders, as well as the avidity effect arising from the multivalent interaction of dimeric hFc with the capture surface, influences the pH dependence of dissociation from the capture surface. Therefore, further studies are needed to evaluate if the Sso7d mutants identified in this study are indeed useful as affinity ligands in chromatography.
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