N-Terminal Modification of Gly-His-Tagged Proteins with Azidogluconolactone.
Chembiochem
; 22(22): 3199-3207, 2021 11 16.
Article
em En
| MEDLINE
| ID: mdl-34520613
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 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.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Azidas
/
Vacinas de Partículas Semelhantes a Vírus
/
Vacinas contra COVID-19
/
Gluconatos
/
Glicina
/
Histidina
/
Lactonas
Limite:
Humans
Idioma:
En
Ano de publicação:
2021
Tipo de documento:
Article