Edwards, Rebecca Louise (2013). Direct surface sampling of dried blood spots coupled with mass spectrometry for haemoglobin analysis. University of Birmingham. Ph.D.
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Edwards13PhD.pdf
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Abstract
Haemoglobinopathies are inherited disorders, typically detected during neonatal healthcare screening programmes. Haemoglobinopathies are characterised by either a reduction in the synthesis of the globin chains or by point mutations in the globin gene often leading to a single amino acid substitution in the globin chain. Current screening techniques analyse samples from resolubilised dried blood spots (DBS) by HPLC and/or isoelectric focusing. These methods are characterised by lengthy sample preparation and/or ambiguous variant determination. Further analysis is required for unequivocal diagnosis. The work presented here describes a method for the unambiguous diagnosis of haemoglobin variants in neonatal DBS samples using a surface sampling technique called liquid extraction surface analysis (LESA) coupled with high resolution top-down mass spectrometry. LESA allows DBS samples to be sampled directly and the material recovered to be directly electrosprayed into a mass spectrometer. Both the sampling and MS/MS (CID/ETD) parameters were optimised to yield maximum globin chain sequence coverage (up to 81 %) allowing for the unambiguous diagnosis of common Hb variants such as HbS and so-called unknown variants (variants unable to be diagnosed during the standard screening protocol). The LESA sampling technique has also successfully been applied to study Hb non-covalent interactions.
Type of Work: | Thesis (Doctorates > Ph.D.) | ||||||
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Award Type: | Doctorates > Ph.D. | ||||||
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College/Faculty: | Colleges (2008 onwards) > College of Life & Environmental Sciences | ||||||
School or Department: | School of Biosciences | ||||||
Funders: | None/not applicable | ||||||
Subjects: | Q Science > QH Natural history > QH301 Biology | ||||||
URI: | http://etheses.bham.ac.uk/id/eprint/4602 |
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