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N-Terminal Modification of Gly-His-Tagged Proteins with Azidogluconolactone

Brune, Karl D. and Liekniņa, Ilva and Sutov, Grigorij and Morris, Alexander R. and Jovicevic, Dejana and Kalniņš, Gints and Kazāks, Andris and Kluga, Rihards and Kastaljana, Sabine and Zajakina, Anna and Jansons, Juris and Skrastiņa, Dace and Spunde, Karīna and Cohen, Alexander A. and Bjorkman, Pamela J. and Morris, Howard R. and Suna, Edgars and Tārs, Kaspars (2021) N-Terminal Modification of Gly-His-Tagged Proteins with Azidogluconolactone. ChemBioChem, 22 (22). pp. 3199-3207. ISSN 1439-4227. doi:10.1002/cbic.202100381.

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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.

Item Type:Article
Related URLs:
URLURL TypeDescription
Brune, Karl D.0000-0003-2796-972X
Kalniņš, Gints0000-0002-9593-6308
Kluga, Rihards0000-0003-1308-9019
Kastaljana, Sabine0000-0002-7700-4371
Zajakina, Anna0000-0002-4753-1103
Jansons, Juris0000-0003-0845-4392
Skrastiņa, Dace0000-0001-5836-6441
Cohen, Alexander A.0000-0002-2818-656X
Bjorkman, Pamela J.0000-0002-2277-3990
Suna, Edgars0000-0002-3078-0576
Tārs, Kaspars0000-0001-8421-9023
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.
Group:COVID-19, Richard N. Merkin Institute for Translational Research
Funding AgencyGrant Number
Latvian Council of ScienceVPP-COVID-2020/1-0014
Lithuanian Agency for Science, Innovation and Technology01.2.1-MITA-T-852-01-0109
Caltech Merkin Institute for Translational ResearchUNSPECIFIED
Genie Biotech Ltd.UNSPECIFIED
Subject Keywords:click chemistry; immunology; nanoparticles; protein modifications; site-specific conjugation
Issue or Number:22
Record Number:CaltechAUTHORS:20211008-224615060
Persistent URL:
Official Citation:N-Terminal Modification of Gly-His-Tagged Proteins with Azidogluconolactone. K. D. Brune, I. Liekniņa, G. Sutov, A. R. Morris, D. Jovicevic, G. Kalniņš, A. Kazāks, R. Kluga, S. Kastaljana, A. Zajakina, J. Jansons, D. Skrastiņa, K. Spunde, A. A. Cohen, P. J. Bjorkman, H. R. Morris, E. Suna, K. Tārs, ChemBioChem 2021, 22, 3199; DOI: 10.1002/cbic.202100381
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:111317
Deposited By: George Porter
Deposited On:12 Oct 2021 20:15
Last Modified:22 Nov 2021 18:14

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