Naslov
High-throughput enrichment of complement C3 protein from human plasma using lectin affinity matrix
(High-throughput enrichment of complement C3 protein from human plasma using lectin affinity matrix)

Autori
Šoić, Dinko ; Keser, Toma ; Novokmet, Mislav ; Lauc, Gordan ; Gornik, Olga

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

Izvornik
27HSKIKI Book of abstracts / - , 2021, 243-243

Skup
27. hrvatski skup kemičara i kemijskih inženjera (27HSKIKI)

Mjesto i datum
Veli Lošinj, Hrvatska, 05.10.2021. - 08.10.2021

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
C3 protein ; Glycopeptides ; High-throughput ; LC-MS ; Lectin affinity ; N-glycosylation
(C3 protein ; glycopeptides ; high-throughput ; LC-MS ; lectin affinity ; N-glycosylation)

Sažetak
The third component of the complement system - glycoprotein C3, plays a central role in the activation of the complement system, both in classical and alternative complement activation pathway. Being the most abundant complement protein with plasma concentrations of approximately 1.2 mg/ml, its position in immune surveillance is pivotal. However, its glycosylation, as one of the most significant post-translation modifications, is not well investigated, but from the literature it is known that C3 has only high mannose N-glycans. Given the growing recognition of complement’s involvement in various disease processes, it is important to have high-throughput methods for the detailed analysis of this key complement protein. The aim of this research was to obtain a C3 enrichment protocol from human plasma for downstream C3 glycopeptide analysis on LC-MS. Knowing the C3 N-glycosylation profile consists of only high mannose structures, glycoprotein enrichment was achieved using lectin affinity matrix. Different volumes of plasma samples and enrichment conditions were tested to get a more specific and effective enrichment. Obtained protocol for C3 enrichment and glycopeptide analysis was applied on 14 healthy male individuals measured in three time points in order to determine the C3 N-glycome stability. This method showed for the first time that the C3 N-glycome is stable in healthy individuals.

Izvorni jezik
Engleski

Znanstvena područja
Farmacija, Biotehnologija u biomedicini (prirodno područje, biomedicina i zdravstvo, biotehničko područje)

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