N-Terminal Modification of Gly-His-Tagged Proteins with Azidogluconolactone
- Creators
- Brune, Karl D.
- Liekniņa, Ilva
- Sutov, Grigorij
- Morris, Alexander R.
- Jovicevic, Dejana
- Kalniņš, Gints
- Kazāks, Andris
- Kluga, Rihards
- Kastaljana, Sabine
- Zajakina, Anna
- Jansons, Juris
- Skrastiņa, Dace
- Spunde, Karīna
- Cohen, Alexander A.
- Bjorkman, Pamela J.
- Morris, Howard R.
- Suna, Edgars
- Tārs, Kaspars
Abstract
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 in vitro α-aminoacylation of peptides, glycoproteins, antibodies, and virus-like particles (VLPs) with azidogluconolactone at pH 7.5 in 1 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.
Additional Information
© 2021 Wiley-VCH. Issue Online: 16 November 2021; Version of Record online: 06 October 2021; Accepted manuscript online: 14 September 2021; Manuscript revised: 13 September 2021; Manuscript received: 30 July 2021. This research was funded by the Latvian Council of Science (VPP-COVID-2020/1-0014), The Lithuanian Agency for Science, Innovation and Technology (MITA, 01.2.1-MITA-T-852-01-0109), the Merkin Institute for Translational Medicine at The California Institute of Technology, BioPharmaSpec Ltd and Genie Biotech Ltd. A.R.M., G.S., and K.D.B. are named inventors on Genie Biotech patent applications for azidogluconoylation. All other authors declare no conflict of interest.Attached Files
Accepted Version - cbic202100381.pdf
Supplemental Material - cbic202100381-sup-0001-misc_information.pdf
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Additional details
- Eprint ID
- 111317
- DOI
- 10.1002/cbic.202100381
- Resolver ID
- CaltechAUTHORS:20211008-224615060
- VPP-COVID-2020/1-0014
- Latvian Council of Science
- 01.2.1-MITA-T-852-01-0109
- Lithuanian Agency for Science, Innovation and Technology
- Caltech Merkin Institute for Translational Research
- BioPharmaSpec Ltd.
- Genie Biotech Ltd.
- Created
-
2021-10-12Created from EPrint's datestamp field
- Updated
-
2023-10-04Created from EPrint's last_modified field
- Caltech groups
- COVID-19, Richard N. Merkin Institute for Translational Research, Division of Biology and Biological Engineering