Utjecaj antikoagulansa korištenog prilikom uzimanja uzoraka krvi na kromatografsku analizu N-glikana plazmatskih proteina

master's thesis

• Title (english): The impact of anticoagulant on chromatographic analysis of N-glycans released from plasma proteins
• Author: Matea Ćurčić
• Mentor: Olga Gornik (mentor)
• Committee member: Olga Gornik (predsjednik povjerenstva)
• Committee member: Nada Vrkić (član povjerenstva)
• Committee member: Suzana Inić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of biochemistry and molecular biology); Zagreb
• Defense date and country: 2016-07-15, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Medical Biochemistry

Abstract

Glikozilacija je najčešća i najvarijabilnija proteinska modifikacija koja ima važnu ulogu u usklađivanju različitih funkcija viših organizama. Glikani osim strukturne imaju i regulatornu ulogu te moduliraju mnoge biološke procese. Upravo zbog velike složenosti i strukturne raznolikosti glikani predstavljaju novu metu znanstvenih istraživanja koja kao uzorak često koriste humanu plazmu. Budući da je u takvim istraživanjima često teško kontrolirati predanalitičke postupke, cilj ovog rada je
bio ispitati utjecaj uzimanja uzorka krvi korištenjem različitih antikoagulansa na rezultate analize Nglikozilacije plazmatskih proteina. Za ostvarivanje navedenog cilja prikupljeni su uzorci plazme ispitanika dobiveni vađenjem krvi na dva različita antikoagulansa, EDTA i Na-citrat. N-glikani su zatim otpušteni s proteina plazme enzimom PNGazom F i obilježeni 2-aminobenzamidom te analizirani tekućinskom kromatografijom visoke djelotvornosti koja se temelji na hidrofilnim interakcijama (HILIC-UPLC kromatografija). Dobiveni rezultati su pokazali da za 25 glikanskih skupina postoji statistički značajna razlika između mjerenja dobivenih nakon uzimanja uzoraka krvi na različiti antikoagulans. Dobivene razlike su male, ali dovoljne da upozore na važnost standardiziranog predanalitičkog postupka. Stoga, da bi interpretacija rezultata laboratorijskih istraživanja bila objektivna i točna, što se od nje i očekuje, nužno je razumijevanje i svjesnost o utjecaju različitih predanalitičkih čimbenika.

Abstract (english)

Glycosylation is the most common and variable protein modification which has important role in harmonizing the various functions of higher organisms. Other than structural, glycans have a regulatory role and modulate numerous biological processes. Due to the great complexity and structural diversity, glycans represent novel targets in scientific research. In this kind of research, it is often very difficult to control preanalytical factors. Therefore, the aim of this study was to examine the
influence of anticoagulant used during blood sampling on the analysis of plasma proteins Nglycosylation. In order to achieve this goal, plasma samples were collected on two types of anticoagulant, EDTA and Na-citrate. Glycans were released using N-glycosidase F, labeled with 2-aminobenzamide and analyzed by high performance liquid chromatography based on hydrophilic interactions (HILICUPLC). Obtained results showed statistically significant difference for 25 glycan groups. Although those differences are small, they show the relevance of standardizing preanalytical procedure for glycan analysis. Therefore, in order to assure accurate interpretation of research results, it is necessary to understand the impact of preanalytical factors.

• Keywords: glikozilacija; N-glikani; predanalitički čimbenici; antikoagulans; HILIC-UPLC
• Keywords (english): glycosylation; N-glycans; preanalytical factors; anticoagulant; HILIC-UPLC
• Language: croatian
• URN:NBN: urn:nbn:hr:163:882018
• Study programme: Title: Medical biochemistry; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra medicinske biokemije (magistar/magistra medicinske biokemije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2016-10-28 11:13:51