6533b856fe1ef96bd12b31d7
RESEARCH PRODUCT
N‐Terminal Modification of Gly‐His‐Tagged Proteins with Azidogluconolactone
Juris JansonsSabine KastaljanaAlexander A. CohenGrigorij SutovPamela J. BjorkmanKarīna SpundeRihards KlugaHoward R. MorrisAlexander R MorrisAndris KazāksKaspars TārsKaspars TārsKarl D. BruneIlva LiekniņaAnna ZajakinaGints KalniņšEdgars SunaDace SkrastiņaDejana Jovicevicsubject
Models MolecularAzidesCOVID-19 VaccinesGlycosylationvirusesGlycineGluconatesBiochemistryLactoneschemistry.chemical_compoundAntigenHumansHistidineVaccines Virus-Like ParticleSeroconversionMolecular Biologychemistry.chemical_classificationMolecular StructurebiologyChemistryOrganic ChemistryAntibodies NeutralizingBiopharmaceuticalBiochemistrybiology.proteinClick chemistryMolecular MedicineAntibodyGlycoproteinConjugatedescription
Site-specific protein modifications are vital for biopharmaceutical drug development. Gluconoylation is a non-enzymatic, post-translational modification of N-terminal HisTags. We report high-yield, site-selective inâ vitro α-aminoacylation of peptides, glycoproteins, antibodies, and virus-like particles (VLPs) with azidogluconolactone at pHâ 7.5 in 1â h. Conjugates slowly hydrolyse, but diol-masking with borate esters inhibits reversibility. In an example, we multimerise azidogluconoylated SARS-CoV-2 receptor-binding domain (RBD) onto VLPs via click-chemistry, to give a COVID-19 vaccine. Compared to yeast antigen, HEK-derived RBD was immunologically superior, likely due to observed differences in glycosylation. We show the benefits of ordered over randomly oriented multimeric antigen display, by demonstrating single-shot seroconversion and best virus-neutralizing antibodies. Azidogluconoylation is simple, fast and robust chemistry, and should accelerate research and development.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2021-11-16 | ChemBioChem |