An engineered redox‐switchable streptavidin mutein enables the high‐affinity capture and efficient elution of biotinylated ligands Article Swipe
Jesse M. Marangoni
,
Adriana Zutic
,
A.S. Dhindsa
,
Emmanuella Uche‐Orji
,
Sui‐Lam Wong
,
Kenneth Ng
·
YOU?
·
· 2025
· Open Access
·
· DOI: https://doi.org/10.1002/pro.70224
YOU?
·
· 2025
· Open Access
·
· DOI: https://doi.org/10.1002/pro.70224
Although the high affinity and specificity of the interaction between streptavidin and biotin are widely utilized to isolate biotinylated ligands, their elution over practical timescales requires the use of strongly denaturing conditions that are incompatible with many target biomolecules. To overcome this limitation, we previously engineered a disulfide bond into a critical loop of streptavidin to create the redox‐sensitive M88 mutein, which releases biotinylated ligands ~19,000‐fold faster in the reduced state when compared with the oxidized state. To optimize the speed and efficiency of ligand recovery, we describe how multiple mechanisms like disulfide bond reduction, increasing temperature, elevating pH, and adding organic cosolvents can dramatically increase the rates of dissociation for different types of biotinylated ligands bound to M88 or wild‐type streptavidin coupled to magnetic beads. By combining these mechanisms, we demonstrate how a range of biotinylated biomolecules (peptide, oligonucleotide, and protein) can be efficiently recovered under mild conditions that preserve biological activity. M88 magnetic beads thus provide many advantages over currently available methods using either wild‐type streptavidin or lower affinity biotin‐binding proteins for applications requiring both high affinity capture and efficient release of target biomolecules under non‐denaturing conditions.
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- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1002/pro.70224
- OA Status
- green
- Cited By
- 1
- References
- 53
- Related Works
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- OpenAlex ID
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All OpenAlex metadata
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https://openalex.org/W4412489210Canonical identifier for this work in OpenAlex
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https://doi.org/10.1002/pro.70224Digital Object Identifier
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An engineered redox‐switchable streptavidin mutein enables the high‐affinity capture and efficient elution of biotinylated ligandsWork title
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articleOpenAlex work type
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enPrimary language
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2025Year of publication
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2025-07-16Full publication date if available
- Authors
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Jesse M. Marangoni, Adriana Zutic, A.S. Dhindsa, Emmanuella Uche‐Orji, Sui‐Lam Wong, Kenneth NgList of authors in order
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https://doi.org/10.1002/pro.70224Publisher landing page
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YesWhether a free full text is available
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greenOpen access status per OpenAlex
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https://www.ncbi.nlm.nih.gov/pmc/articles/12267106Direct OA link when available
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Biotinylation, Streptavidin, Elution, Chemistry, Redox, Biophysics, Combinatorial chemistry, Biotin, Chromatography, Biochemistry, Inorganic chemistry, BiologyTop concepts (fields/topics) attached by OpenAlex
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1Total citation count in OpenAlex
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2025: 1Per-year citation counts (last 5 years)
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10Other works algorithmically related by OpenAlex
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