Naslov
Semi-high-throughput isolation and N-glycan analysis of human fibrinogen using monolithic supports bearing monoclonal anti-human fibrinogen antibodies
(Semi-high-throughput isolation and N -glycan analysis of human fibrinogen using monolithic supports bearing monoclonal anti-human fibrinogen antibodies)

Autori
Vidic, Urška ; Trbojević-Akmačić, Irena ; Černigoj, Urh ; Albers, Malena ; Gašperšič, Jernej ; Pučić-Baković, Maja ; Vidič, Jana ; Štrancar, Aleš ; Lauc, Gordan

Izvornik
ELECTROPHORESIS (0173-0835) 38 (2017), 22-23; 2922-2930

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
Fibrinogen ; High-throughput ; Immunoaffinity chromatography ; Monoliths ; N-glycosylation

Sažetak
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.

Izvorni jezik
Engleski

Znanstvena područja
Kemija, Biologija, Biotehnologija

POVEZANOST RADA