INVESTIGADORES
MARIÑO Karina Valeria
convenios, asesorías y/o servicios tecnológicos
Título:
Análisis de glicosilación de glicoproteína recombinante
Autor/es:
MARIÑO, KARINA V; CAGNONI, ALEJANDRO J.
Fecha inicio:
2019-07-01
Fecha finalización:
2019-08-01
Campo de Aplicación:
Otros campos
Descripción:
Due to its complex biosynthesis process, glycoproteins are naturally heterogeneous, generating different specific glycoforms. Glycoanalytics nowadays have several technologies available for characterization of a specific glycosylation profile. Among them, N-glycan characterization after fluorescent derivatization by chromatographic techniques is one of the most used.For the analysis of the N-glycans in this glycoprotein heterologously expressed in Leishmania tarentolae, the enzymatic release of glycans and their subsequent fluorescent labeling is required (Royle et al., 2008, Mariño et al., 2010). In this analysis we used an Acquity H-Class (Waters) equipment coupled to a fluorescence detector. N-glycans were released by an endoglycanase (PNGase F) and were labeled with a fluorescent probe (2-aminobenzamide, 2-AB), which makes the fluorescent signal independent of the glycan structure and allows relative quantification (Royle et al., 2008). HILIC-UPLC-FLR methodology was applied for the separation and characterization of profiles.
