SpySwitch enables pH- or heat-responsive capture and release for plug-and-display nanoassembly

Susan K. Vester, Rolle Rahikainen, Irsyad N.A. Khairil Anuar, Rory A. Hills, Tiong Kit Tan, Mark Howarth

    Research output: Contribution to journalArticleScientificpeer-review

    21 Citations (Scopus)
    29 Downloads (Pure)

    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.

    Original languageEnglish
    Article number3714
    JournalNature Communications
    Volume13
    Issue number1
    DOIs
    Publication statusPublished - Dec 2022
    Publication typeA1 Journal article-refereed

    Publication forum classification

    • Publication forum level 3

    ASJC Scopus subject areas

    • General Chemistry
    • General Biochemistry,Genetics and Molecular Biology
    • General
    • General Physics and Astronomy

    Fingerprint

    Dive into the research topics of 'SpySwitch enables pH- or heat-responsive capture and release for plug-and-display nanoassembly'. Together they form a unique fingerprint.

    Cite this