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SpySwitch enables pH- or heat-responsive capture and release for plug-and-display nanoassembly.
Vester, Susan K; Rahikainen, Rolle; Khairil Anuar, Irsyad N A; Hills, Rory A; Tan, Tiong Kit; Howarth, Mark.
  • Vester SK; Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK.
  • Rahikainen R; Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK.
  • Khairil Anuar INA; Faculty of Medicine and Health Technology, Tampere University, 33014, Tampere, Finland.
  • Hills RA; Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK.
  • Tan TK; LiliumX Ltd, WE.306 Westbourne Studios, 242 Acklam Road, London, W10 5JJ, UK.
  • Howarth M; Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK.
Nat Commun ; 13(1): 3714, 2022 06 28.
Article in English | MEDLINE | ID: covidwho-1908171
ABSTRACT
Proteins can be empowered via SpyTag for anchoring and nanoassembly, through covalent bonding to SpyCatcher partners. Here we generate a switchable version of SpyCatcher, allowing gentle purification of SpyTagged proteins. We introduce numerous histidines adjacent to SpyTag's binding site, giving moderate pH-dependent release. After phage-based selection, our final SpySwitch allows purification of SpyTag- and SpyTag003-fusions from bacterial or mammalian culture by capture at neutral pH and release at pH 5, with purity far beyond His-tag methods. SpySwitch is also thermosensitive, capturing at 4 °C and releasing at 37 °C. With flexible choice of eluent, SpySwitch-purified proteins can directly assemble onto multimeric scaffolds. 60-mer multimerization enhances immunogenicity and we use SpySwitch to purify receptor-binding domains from SARS-CoV-2 and 11 other sarbecoviruses. For these receptor-binding domains we determine thermal resilience (for mosaic vaccine development) and cross-recognition by antibodies. Antibody EY6A reacts across all tested sarbecoviruses, towards potential application against new coronavirus pandemic threats.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: COVID-19 / Hot Temperature Type of study: Randomized controlled trials Topics: Vaccines Limits: Animals Language: English Journal: Nat Commun Journal subject: Biology / Science Year: 2022 Document Type: Article Affiliation country: S41467-022-31193-8

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Full text: Available Collection: International databases Database: MEDLINE Main subject: COVID-19 / Hot Temperature Type of study: Randomized controlled trials Topics: Vaccines Limits: Animals Language: English Journal: Nat Commun Journal subject: Biology / Science Year: 2022 Document Type: Article Affiliation country: S41467-022-31193-8