Artículo
Bifunctional GM-CSF-derived peptides as tools for O-glycoengineering and protein tagging
Sales, María de Los Milagros; Kratje, Ricardo Bertoldo
; Oggero Eberhardt, Marcos Rafael
; Ceaglio, Natalia Analia
Fecha de publicación:
02/2021
Editorial:
Elsevier Science
Revista:
Journal of Biotechnology
ISSN:
0168-1656
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
Rapid development of effective biotherapeutics has been a concern during the last couple decades. In our work we designed two novel peptide tags, GMOP and mGMOP, derived from the N-terminal region of human granulocyte and macrophage colony stimulating factor (hGM-CSF), which contain four and six potential O-glycosylation sites, respectively. These peptide tags were fused to the N-terminus of human interferon-α2b (hIFN-α2b), a therapeutic antiviral and antiproliferative protein rapidly cleared from circulation. Two new molecules were obtained which, consistently with the presence of O-glycans, showed higher molecular masses, more negatively charged isoforms, and higher sialic acid content compared to wild-type IFN. In vitro bioactivity of purified chimeras revealed a similar antiviral specific biological activity (SBA) compared to unmodified IFN. A reduction of antiproliferative SBA was only observed for mGMOP-IFN. Pharmacokinetic studies in rats showed a notable improvement in terminal half-life (t1/2elim) (3.3 and 2.8 times-longer) and a marked reduction of the apparent clearance (CLapp, 3.7 and 4.1-fold lower for GMOP-IFN and mGMOP-IFN in comparison with native IFN, respectively). Furthermore, the in vitro thermal and plasma stability of both proteins was improved. Finally, a monoclonal antibody (mAb) that recognizes an N-terminal GM-CSF epitope was able to bind both chimeras in western blots and ELISAs. This demonstrates the potential of both peptides to behave as bifunctional tags to create novel long-acting biotherapeutics and to facilitate detection and purification.
Palabras clave:
FUSION PROTEINS
,
O-GLYCOSYLATION
,
PEPTIDE TAGGING
,
PHARMACOKINETICS
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Colecciones
Articulos(CCT - SANTA FE)
Articulos de CTRO.CIENTIFICO TECNOL.CONICET - SANTA FE
Articulos de CTRO.CIENTIFICO TECNOL.CONICET - SANTA FE
Citación
Sales, María de Los Milagros; Kratje, Ricardo Bertoldo; Oggero Eberhardt, Marcos Rafael; Ceaglio, Natalia Analia; Bifunctional GM-CSF-derived peptides as tools for O-glycoengineering and protein tagging; Elsevier Science; Journal of Biotechnology; 327; 2-2021; 18-27
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