Naslov
Mass spectrometry identification of recombinant human erythropoietin N-glycans utilizing a novel automated method

Autori
Dončević, Lucija ; Svetličić, Ema ; Ozdanovac, Luka ; Cindrić, Mario

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Skup
4th International conference on post-translational modifications in Bacteria

Mjesto i datum
Kopenhagen, Danska, 05.05.2022. - 06.05.2022

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
post-translational modifications ; glycosylation ; recombinant human erythropoetin ; MALDI-TOF-MS

Sažetak
The analysis of post-translational modifications is one of the main goals of modern protein structure/activity research, especially glycosylation structure elucidation. To understand N- and O-glycoprotein function and the role of protein glycosylation, the fundamental prerequisite is the detailed structural elucidation of both protein and its glycan moieties. Glycosylation is widely used to increase protein stability, cell adhesion, and proper protein folding and immunogenicity. For this purpose, therapeutic protein drugs are often glycosylated, which increases the need for better and faster glycan studies. The frequently used drug recombinant human erythropoietin (rHuEPO) is a highly glycosylated hormone used to treat various diseases such as anemia, chronic renal disease, hematological disorders, cancer, etc. Glycan sample preparation, the most important step in glycan analysis, often includes a combination of two or more chromatographic methods. For identification purposes, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) is often used for the analysis of the N-glycan mixture. In this study, we have automated sample preparation by using a graphitized carbon column on the PosiTip Automated liquid handling platform for the purification and fractionation of N-glycans. The automated method enables the identification of an N-glycan mixture of up to 96 samples simultaneously under 24 hours with minimal sample loss.

Izvorni jezik
Engleski

Znanstvena područja
Kemija

POVEZANOST RADA