Please use this identifier to cite or link to this item: http://scholarbank.nus.edu.sg/handle/10635/139074
Title: BIOSYNTHESIS, ISOLATION AND CHARACTERISATION OF ISOTOPICALLY LABELLED SPIKES FOR FERRITIN ANALYSIS BY ISOTOPE DILUTION MASS SPECTROMETRY
Authors: DORRY POA MING HIANG
Keywords: Biosynthesis, Isolation, Characterisation, Ferritin, Isotope, Labelled
Issue Date: 25-Aug-2017
Citation: DORRY POA MING HIANG (2017-08-25). BIOSYNTHESIS, ISOLATION AND CHARACTERISATION OF ISOTOPICALLY LABELLED SPIKES FOR FERRITIN ANALYSIS BY ISOTOPE DILUTION MASS SPECTROMETRY. ScholarBank@NUS Repository.
Abstract: Ferritin is found in most living organism and is responsible for maintaining normal physiological conditions in the human body through iron storage. Serum ferritin concentration is currently the most specific and best single indicator to assess body iron stores. Although current techniques used to quantify serum ferritin are simple and rapid, several limitations exist. One of the biggest concerns of traditional quantification techniques lies in the inability to achieve traceable results metrologically to the SI system of units. This restricts the comparison of results across time and space. . In this study, recombinant human L-chain and H-chain ferritin were successfully cultured, labelled with nitrogen-15 (15N) and purified for ferritin analysis by isotope dilution mass spectrometry. Two proteotypic peptides per ferritin chain were identified through mass spectrometric techniques and purchased to characterise 15N labelled ferritin for concentration and isotopic composition. Before quantification experiments, mass spectrometric isotopologue abundance patterns were validated, whereby repeatable isotopologue ratios were achieved across different concentrations and days. Biosynthesised 15N labelled ferritins were quantified using the technique of organic isotope dilution using unlabelled proteotypic peptides as spike.
URI: http://scholarbank.nus.edu.sg/handle/10635/139074
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