Characterization of airborne allergens of selected crustaceans and monitoring of their occupational exposure levels using mass spectrometry
Thesis (Ph.D.)--Memorial University of Newfoundland, 2011. Chemistry Includes bibliographical references. The number of workers in the processing and harvesting sector of the fishery has increased substantially in recent decades, seeing more than 41 million workers worldwide engaged in various activ...
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ftmemorialunivdc:oai:collections.mun.ca:theses5/39254 2023-05-15T17:23:28+02:00 Characterization of airborne allergens of selected crustaceans and monitoring of their occupational exposure levels using mass spectrometry Abdel Rahman, Anas M. (Anas Mahmoud) Memorial University of Newfoundland. Dept. of Chemistry 2011 xix, 191 leaves : ill. Image/jpeg; Application/pdf http://collections.mun.ca/cdm/ref/collection/theses5/id/39254 Eng eng Electronic Theses and Dissertations (20.80 MB) -- http://collections.mun.ca/PDFs/theses/Rahman_AnasMAbdel.pdf http://collections.mun.ca/cdm/ref/collection/theses5/id/39254 The author retains copyright ownership and moral rights in this thesis. Neither the thesis nor substantial extracts from it may be printed or otherwise reproduced without the author's permission. Paper copy kept in the Centre for Newfoundland Studies, Memorial University Libraries Allergens--Testing Proteins--Allergenicity Mass spectrometry Shellfish fisheries Fishery processing industries Respiratory allergy Text Electronic thesis or dissertation 2011 ftmemorialunivdc 2015-08-06T19:22:53Z Thesis (Ph.D.)--Memorial University of Newfoundland, 2011. Chemistry Includes bibliographical references. The number of workers in the processing and harvesting sector of the fishery has increased substantially in recent decades, seeing more than 41 million workers worldwide engaged in various activities related to seafood production. Serious allergies and asthma have developed in the seafood occupational workplace as the result of airborne allergenic proteins being aerosolized on fishing vessels and in processing plants. Traditionally, the identification and quantification of these allergens are performed by immunoactive-based techniques such as radioallergosorbent test and enzyme-linked immunosorbent assay. -- In this study, a novel strategy of monitoring airborne seafood allergens was developed using tandem mass spectrometry. First, the major allergenic proteins of snow crab and black tiger prawn were identified and profiled using an allergeomics approach. The immunoblotting was performed against sensitized patients' sera and the active protein bands were characterized using peptide mass fingerprinting mass spectrometry. The purified forms of the target allergenic proteins, tropomyosin and arginine kinase, were used as reference materials for primary structural identification and for quantification method development. -- For protein sequencing, the samples were exposed to different enzymatic digestions (Trypsin, Glu-C V8, and Asp-N), the peptides analyzed by different mass spectrometry ion sources, and various derivatization protocols were used to maximize the amino acid sequence coverage. The product mass spectra of the enzymatically produced peptides were uploaded to a Mascot search engine and the national center for biotechnology information non-redundant database searched. Signature peptides that surrogate each individual allergenic protein were determined and chemically synthesized (purchased from third party) for use as analytical standards in both light and heavy forms. Subsequently, a quantification approach using liquid chromatography-multiple reaction monitoring isotopic dilution tandem mass spectrometry was developed to measure snow crab tropomyosin and arginine kinase in air samples. A comprehensive validation study was performed for the quantification method with respect to the international conference on harmonization guidelines. The air sampling protocol, allergen extraction recovery, and the completeness of the protein digestion were optimized for real air sample collection. Actual air samples were collected from a simulated crab processing plant and analyzed by mass spectrometry. Thesis Newfoundland studies Snow crab University of Newfoundland Memorial University of Newfoundland: Digital Archives Initiative (DAI) |
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Memorial University of Newfoundland: Digital Archives Initiative (DAI) |
op_collection_id |
ftmemorialunivdc |
language |
English |
topic |
Allergens--Testing Proteins--Allergenicity Mass spectrometry Shellfish fisheries Fishery processing industries Respiratory allergy |
spellingShingle |
Allergens--Testing Proteins--Allergenicity Mass spectrometry Shellfish fisheries Fishery processing industries Respiratory allergy Abdel Rahman, Anas M. (Anas Mahmoud) Characterization of airborne allergens of selected crustaceans and monitoring of their occupational exposure levels using mass spectrometry |
topic_facet |
Allergens--Testing Proteins--Allergenicity Mass spectrometry Shellfish fisheries Fishery processing industries Respiratory allergy |
description |
Thesis (Ph.D.)--Memorial University of Newfoundland, 2011. Chemistry Includes bibliographical references. The number of workers in the processing and harvesting sector of the fishery has increased substantially in recent decades, seeing more than 41 million workers worldwide engaged in various activities related to seafood production. Serious allergies and asthma have developed in the seafood occupational workplace as the result of airborne allergenic proteins being aerosolized on fishing vessels and in processing plants. Traditionally, the identification and quantification of these allergens are performed by immunoactive-based techniques such as radioallergosorbent test and enzyme-linked immunosorbent assay. -- In this study, a novel strategy of monitoring airborne seafood allergens was developed using tandem mass spectrometry. First, the major allergenic proteins of snow crab and black tiger prawn were identified and profiled using an allergeomics approach. The immunoblotting was performed against sensitized patients' sera and the active protein bands were characterized using peptide mass fingerprinting mass spectrometry. The purified forms of the target allergenic proteins, tropomyosin and arginine kinase, were used as reference materials for primary structural identification and for quantification method development. -- For protein sequencing, the samples were exposed to different enzymatic digestions (Trypsin, Glu-C V8, and Asp-N), the peptides analyzed by different mass spectrometry ion sources, and various derivatization protocols were used to maximize the amino acid sequence coverage. The product mass spectra of the enzymatically produced peptides were uploaded to a Mascot search engine and the national center for biotechnology information non-redundant database searched. Signature peptides that surrogate each individual allergenic protein were determined and chemically synthesized (purchased from third party) for use as analytical standards in both light and heavy forms. Subsequently, a quantification approach using liquid chromatography-multiple reaction monitoring isotopic dilution tandem mass spectrometry was developed to measure snow crab tropomyosin and arginine kinase in air samples. A comprehensive validation study was performed for the quantification method with respect to the international conference on harmonization guidelines. The air sampling protocol, allergen extraction recovery, and the completeness of the protein digestion were optimized for real air sample collection. Actual air samples were collected from a simulated crab processing plant and analyzed by mass spectrometry. |
author2 |
Memorial University of Newfoundland. Dept. of Chemistry |
format |
Thesis |
author |
Abdel Rahman, Anas M. (Anas Mahmoud) |
author_facet |
Abdel Rahman, Anas M. (Anas Mahmoud) |
author_sort |
Abdel Rahman, Anas M. (Anas Mahmoud) |
title |
Characterization of airborne allergens of selected crustaceans and monitoring of their occupational exposure levels using mass spectrometry |
title_short |
Characterization of airborne allergens of selected crustaceans and monitoring of their occupational exposure levels using mass spectrometry |
title_full |
Characterization of airborne allergens of selected crustaceans and monitoring of their occupational exposure levels using mass spectrometry |
title_fullStr |
Characterization of airborne allergens of selected crustaceans and monitoring of their occupational exposure levels using mass spectrometry |
title_full_unstemmed |
Characterization of airborne allergens of selected crustaceans and monitoring of their occupational exposure levels using mass spectrometry |
title_sort |
characterization of airborne allergens of selected crustaceans and monitoring of their occupational exposure levels using mass spectrometry |
publishDate |
2011 |
url |
http://collections.mun.ca/cdm/ref/collection/theses5/id/39254 |
genre |
Newfoundland studies Snow crab University of Newfoundland |
genre_facet |
Newfoundland studies Snow crab University of Newfoundland |
op_source |
Paper copy kept in the Centre for Newfoundland Studies, Memorial University Libraries |
op_relation |
Electronic Theses and Dissertations (20.80 MB) -- http://collections.mun.ca/PDFs/theses/Rahman_AnasMAbdel.pdf http://collections.mun.ca/cdm/ref/collection/theses5/id/39254 |
op_rights |
The author retains copyright ownership and moral rights in this thesis. Neither the thesis nor substantial extracts from it may be printed or otherwise reproduced without the author's permission. |
_version_ |
1766112477491232768 |