Semi-high-throughput isolation and N-glycan analysis of human fibrinogen using monolithic supports bearing monoclonal anti-human fibrinogen antibodies.
Electrophoresis
; 38(22-23): 2922-2930, 2017 11.
Article
en En
| MEDLINE
| ID: mdl-28556996
ABSTRACT
Fibrinogen (FIB) is a secretory glycoprotein synthesized by hepatocytes that has a key role in blood clotting. Its glycosylation has not been studied in detail and little is known about the biological variability of FIB N-glycosylation, mainly due to the lack of fast, simple, and robust approaches to purify FIB from blood plasma samples. In recent years, customised chromatographic monoliths have been used for a variety of biological applications due to their unique characteristics. Here we describe development and optimisation of monolithic supports bearing monoclonal anti-human fibrinogen antibodies in a single column as well as in multi-well plate formats with high FIB specificity and binding capacity for fast immunoaffinity purification of FIB from human blood samples. The developed semi-high-throughput workflow has been successfully applied for FIB immunoaffinity isolation and subsequent ultra performance liquid chromatography N-glycosylation analysis in ten healthy human individuals, demonstrating the potential of monolithic supports in glycomics studies.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Fibrinógeno
/
Cromatografía de Afinidad
/
Anticuerpos Inmovilizados
/
Ensayos Analíticos de Alto Rendimiento
/
Anticuerpos Monoclonales
Límite:
Humans
Idioma:
En
Revista:
Electrophoresis
Año:
2017
Tipo del documento:
Article
País de afiliación:
Eslovenia