Quantitation of enteroendocrine hormones as metabolic disease biomarkers in human biological matrices by liquid chromatography tandem mass spectrometry
type: CreativeWork
description: Polypeptides are widely present in biology and many of them act as biomarkers, both for disease and system regulation. Many gut hormone peptides are secreted by enteroendocrine cells (EECs) and some, known as incretins, are released after a meal to control blood glucose and stimulate neuronal satiety. These are being studied as novel targets for treatment of metabolic diseases such as obesity. Traditionally quantitative analysis of these hormones is performed by enzyme-linked immunosorbent assays (ELISAs). The development of these immunological techniques can take many years and is not always cost effective, so new biomarker assay approaches are being explored in research. Liquid Chromatography Mass Spectrometry (LC-MS/MS) methods are becoming an alternative approach and analytical technique combines the physical separation abilities of liquid chromatography with specific mass analysis of mass spectrometry. The methods described in this thesis are traditional bioanalytical extractions used to isolate small molecule analytes from biological matrices. Assay development can be a challenging process, especially when the target biomarker is expected to be in the low (picomolar) ranges. To improve sensitivity of the methods a number of different extraction procedures were investigated and the optimal conditions for each peptide target were identified. Additionally the mass spectrometry hardware systems were assessed and modified, working to find a balance between sensitive detection limits and reducing sample analysis time to maintain high throughput methods for clinical application. Where applicable, the optimised LC-MS/MS methods were validated by commercial immunoassays and the strong correlation in concentrations confirmed the accuracy of the developed assay. An additional advantage of mass spectrometry is the ability to monitor multiple analytes in a single sample, so additional data on all possible isoforms of the target analyte are available, as opposed to the ‘tot…
abstract: Polypeptides are widely present in biology and many of them act as biomarkers, both for disease and system regulation. Many gut hormone peptides are secreted by enteroendocrine cells (EECs) and some, known as incretins, are released after a meal to control blood glucose and stimulate neuronal satiety. These are being studied as novel targets for treatment of metabolic diseases such as obesity. Traditionally quantitative analysis of these hormones is performed by enzyme-linked immunosorbent assays (ELISAs). The development of these immunological techniques can take many years and is not always cost effective, so new biomarker assay approaches are being explored in research. Liquid Chromatography Mass Spectrometry (LC-MS/MS) methods are becoming an alternative approach and analytical technique combines the physical separation abilities of liquid chromatography with specific mass analysis of mass spectrometry. The methods described in this thesis are traditional bioanalytical extractions used to isolate small molecule analytes from biological matrices. Assay development can be a challenging process, especially when the target biomarker is expected to be in the low (picomolar) ranges. To improve sensitivity of the methods a number of different extraction procedures were investigated and the optimal conditions for each peptide target were identified. Additionally the mass spectrometry hardware systems were assessed and modified, working to find a balance between sensitive detection limits and reducing sample analysis time to maintain high throughput methods for clinical application. Where applicable, the optimised LC-MS/MS methods were validated by commercial immunoassays and the strong correlation in concentrations confirmed the accuracy of the developed assay. An additional advantage of mass spectrometry is the ability to monitor multiple analytes in a single sample, so additional data on all possible isoforms of the target analyte are available, as opposed to the ‘tot…
author: Foreman, Rachel
language: eng
keywords: bioanalysis, enteroendocrine, mass spectrometry, metabolic diseases, peptides
license: https://www.repository.cam.ac.uk/bitstreams/617a0b0f-2183-4d33-9ae9-9bc6ee62917c/download
license: https://www.rioxx.net/licenses/all-rights-reserved/
date published: 2023-03-01
date modified: 2026-07-24T01:33:11Z
provider: Cambridge University
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: University of Cambridge

PropertyValue
property ID: DOI
value: 10.17863/CAM.102080