{"id":{"repo_id":"southwales","oai_identifier":"oai:pure.atira.dk:studenttheses/2f7b4682-04e3-44f4-a56f-25760683e7ab"},"canonical_url":"https://search.dev.ndltd.org/etd/southwales/oai:pure.atira.dk:studenttheses/2f7b4682-04e3-44f4-a56f-25760683e7ab","repository":{"repo_id":"southwales","name":"University of South Wales","base_url":"https://pure.southwales.ac.uk/ws/oai"},"display":{"title":"Development of a Chemiluminescent Assay for Autoantibodies to Thyroid Peroxidase","abstract":"The presence of antibodies to thyroid peroxidase (i.e. TPOAb) in human serum has been measured using an immunoradiometric assay (IRMA), an enzyme-immunometric assay, and three other immunoassay systems which incorporated the chemiluminescent label acridinium ester (AE).
The IRMA successfully utilised a commercial preparation of TPO labelled with [125I] and magnetic particles coupled to Protein A (Staphylococcus aureus) (MPA), with the latter separation medium used in an enzyme-immunometric assay which incorporated TPO labelled with horse-radish peroxidase (HRP-TPO). TPO was also labelled with AE and used in a chemiluminometric assay with magnetic particles coupled to human anti-IgG (MAb-Anti-IgG). The latter assay exhibited an improved response, as compared with the non-viable enzyme-immunometric assay, but was inferior to the IRMA lacking the required sensitivity and precision for a viable, clinical assay.
A competitive assay system was also investigated which utilised human anti-TPO (obtained from a purified preparation of IgG) labelled with acridinium ester (Acrid-IgG), TPO labelled with biotin (Biotin-TPO) and magnetic streptavidin-labelled 'Dynabeads®', as the separation medium. This immunoassay produced a similar 'blanket' response as produced in the enzyme-immunometric assay. The coated-tube assay involved the immobilisation of TPO onto plastic tubes and the use of a commercial preparation of sheep anti-human IgG labelled with AE (Acrid-Anti-IgG). This system proved to be the most viable of all the non-radioactive assays, with the results comparing favourably with the established ELISA with a correlation coefficient (r) of 0.96 and P = <0.001, (using the 'least squares linear regression after logarithmic conversion of the data). A good correlation was also demonstrated in the Deming and the Passing & Bablok plots. The coated-tube assay also compared favourably with indirect agglutination (r) = 0.80, P = <0.001.
The results indicated that the less pure source of TPO used in the non-radioactive assays, could only be successfully applied to the measurement of TPOAb, when immobilised onto a solid-phase support such as the ELISA micro-titre plate or onto plastic tubes, but not in the more random, assay systems which utilised magnetic particles as the separation phase. The solid-phase, coated-tube chemiluminometric assay was comparable with ELISA for the measurement of TPOAb in human serum.","abstract_html":"The presence of antibodies to thyroid peroxidase (i.e. TPOAb) in human serum has been measured using an immunoradiometric assay (IRMA), an enzyme-immunometric assay, and three other immunoassay systems which incorporated the chemiluminescent label acridinium ester (AE).<br/><br/>The IRMA successfully utilised a commercial preparation of TPO labelled with [<sup>125</sup>I] and magnetic particles coupled to Protein A (<i>Staphylococcus aureus</i>) (MPA), with the latter separation medium used in an enzyme-immunometric assay which incorporated TPO labelled with horse-radish peroxidase (HRP-TPO). TPO was also labelled with AE and used in a chemiluminometric assay with magnetic particles coupled to human anti-IgG (MAb-Anti-IgG). The latter assay exhibited an improved response, as compared with the non-viable enzyme-immunometric assay, but was inferior to the IRMA lacking the required sensitivity and precision for a viable, clinical assay.<br/><br/>A competitive assay system was also investigated which utilised human anti-TPO (obtained from a purified preparation of IgG) labelled with acridinium ester (Acrid-IgG), TPO labelled with biotin (Biotin-TPO) and magnetic streptavidin-labelled 'Dynabeads®', as the separation medium. This immunoassay produced a similar 'blanket' response as produced in the enzyme-immunometric assay. The coated-tube assay involved the immobilisation of TPO onto plastic tubes and the use of a commercial preparation of sheep anti-human IgG labelled with AE (Acrid-Anti-IgG). This system proved to be the most viable of all the non-radioactive assays, with the results comparing favourably with the established ELISA with a correlation coefficient (r) of 0.96 and P = <0.001, (using the 'least squares linear regression after logarithmic conversion of the data). A good correlation was also demonstrated in the Deming and the Passing & Bablok plots. The coated-tube assay also compared favourably with indirect agglutination (r) = 0.80, P = <0.001.<br/><br/>The results indicated that the less pure source of TPO used in the non-radioactive assays, could only be successfully applied to the measurement of TPOAb, when immobilised onto a solid-phase support such as the ELISA micro-titre plate or onto plastic tubes, but not in the more random, assay systems which utilised magnetic particles as the separation phase. The solid-phase, coated-tube chemiluminometric assay was comparable with ELISA for the measurement of TPOAb in human serum.","abstract_has_math":false,"creators":["Thomas, Karen"],"institution":null,"degree_name":"Master's Thesis","degree_level":"Student thesis","degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2000,"date_issued":"2000-7","date_published":"2000-7","updated_at":"2026-07-24T04:38:59Z","subjects":["Biochemistry","Autoantibodies","Thyroid Peroxida","TPOAb","immunoradiometric assay","IRMA","enzymeimmunometric assay","Chemiluminescent Assay","acridinium ester (AE)"],"languages":["eng"],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["oai:pure.atira.dk:studenttheses/2f7b4682-04e3-44f4-a56f-25760683e7ab"],"render_values":[{"text":"oai:pure.atira.dk:studenttheses/2f7b4682-04e3-44f4-a56f-25760683e7ab","href":null,"code":true}]}]},"links":{"outbound_url":"https://pure.southwales.ac.uk/en/studentTheses/2f7b4682-04e3-44f4-a56f-25760683e7ab","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:creator","label":"Author","values":["Thomas, Karen"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2000-7"]},{"key":"dc:date.issued","label":"Date","values":["2000-7"]},{"key":"dc:relation.isreferencedby","label":"Dc Relation Isreferencedby","values":["https://pure.southwales.ac.uk/en/studentTheses/2f7b4682-04e3-44f4-a56f-25760683e7ab"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]},{"key":"dc:type.qualificationlevel","label":"Dc Type Qualificationlevel","values":["Student thesis"]},{"key":"dc:type.qualificationname","label":"Dc Type Qualificationname","values":["Master's Thesis"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Biochemistry","Autoantibodies","Thyroid Peroxida","TPOAb","immunoradiometric assay","IRMA","enzymeimmunometric assay","Chemiluminescent Assay","acridinium ester (AE)"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["oai:pure.atira.dk:studenttheses/2f7b4682-04e3-44f4-a56f-25760683e7ab","https://pure.southwales.ac.uk/en/studentTheses/2f7b4682-04e3-44f4-a56f-25760683e7ab"]},{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://pure.southwales.ac.uk/files/2104139/K._C._Thomas_2000_2059454.pdf"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["The presence of antibodies to thyroid peroxidase (i.e. TPOAb) in human serum has been measured using an immunoradiometric assay (IRMA), an enzyme-immunometric assay, and three other immunoassay systems which incorporated the chemiluminescent label acridinium ester (AE).
The IRMA successfully utilised a commercial preparation of TPO labelled with [125I] and magnetic particles coupled to Protein A (Staphylococcus aureus) (MPA), with the latter separation medium used in an enzyme-immunometric assay which incorporated TPO labelled with horse-radish peroxidase (HRP-TPO). TPO was also labelled with AE and used in a chemiluminometric assay with magnetic particles coupled to human anti-IgG (MAb-Anti-IgG). The latter assay exhibited an improved response, as compared with the non-viable enzyme-immunometric assay, but was inferior to the IRMA lacking the required sensitivity and precision for a viable, clinical assay.
A competitive assay system was also investigated which utilised human anti-TPO (obtained from a purified preparation of IgG) labelled with acridinium ester (Acrid-IgG), TPO labelled with biotin (Biotin-TPO) and magnetic streptavidin-labelled 'Dynabeads®', as the separation medium. This immunoassay produced a similar 'blanket' response as produced in the enzyme-immunometric assay. The coated-tube assay involved the immobilisation of TPO onto plastic tubes and the use of a commercial preparation of sheep anti-human IgG labelled with AE (Acrid-Anti-IgG). This system proved to be the most viable of all the non-radioactive assays, with the results comparing favourably with the established ELISA with a correlation coefficient (r) of 0.96 and P = <0.001, (using the 'least squares linear regression after logarithmic conversion of the data). A good correlation was also demonstrated in the Deming and the Passing & Bablok plots. The coated-tube assay also compared favourably with indirect agglutination (r) = 0.80, P = <0.001.
The results indicated that the less pure source of TPO used in the non-radioactive assays, could only be successfully applied to the measurement of TPOAb, when immobilised onto a solid-phase support such as the ELISA micro-titre plate or onto plastic tubes, but not in the more random, assay systems which utilised magnetic particles as the separation phase. The solid-phase, coated-tube chemiluminometric assay was comparable with ELISA for the measurement of TPOAb in human serum."]},{"key":"dc:title","label":"Title","values":["Development of a Chemiluminescent Assay for Autoantibodies to Thyroid Peroxidase"]}]}],"canonical_facts":{"dc:creator":["Thomas, Karen"],"dc:date":["2000-7"],"dc:date.issued":["2000-7"],"dc:description.abstract":["The presence of antibodies to thyroid peroxidase (i.e. TPOAb) in human serum has been measured using an immunoradiometric assay (IRMA), an enzyme-immunometric assay, and three other immunoassay systems which incorporated the chemiluminescent label acridinium ester (AE).
The IRMA successfully utilised a commercial preparation of TPO labelled with [125I] and magnetic particles coupled to Protein A (Staphylococcus aureus) (MPA), with the latter separation medium used in an enzyme-immunometric assay which incorporated TPO labelled with horse-radish peroxidase (HRP-TPO). TPO was also labelled with AE and used in a chemiluminometric assay with magnetic particles coupled to human anti-IgG (MAb-Anti-IgG). The latter assay exhibited an improved response, as compared with the non-viable enzyme-immunometric assay, but was inferior to the IRMA lacking the required sensitivity and precision for a viable, clinical assay.
A competitive assay system was also investigated which utilised human anti-TPO (obtained from a purified preparation of IgG) labelled with acridinium ester (Acrid-IgG), TPO labelled with biotin (Biotin-TPO) and magnetic streptavidin-labelled 'Dynabeads®', as the separation medium. This immunoassay produced a similar 'blanket' response as produced in the enzyme-immunometric assay. The coated-tube assay involved the immobilisation of TPO onto plastic tubes and the use of a commercial preparation of sheep anti-human IgG labelled with AE (Acrid-Anti-IgG). This system proved to be the most viable of all the non-radioactive assays, with the results comparing favourably with the established ELISA with a correlation coefficient (r) of 0.96 and P = <0.001, (using the 'least squares linear regression after logarithmic conversion of the data). A good correlation was also demonstrated in the Deming and the Passing & Bablok plots. The coated-tube assay also compared favourably with indirect agglutination (r) = 0.80, P = <0.001.
The results indicated that the less pure source of TPO used in the non-radioactive assays, could only be successfully applied to the measurement of TPOAb, when immobilised onto a solid-phase support such as the ELISA micro-titre plate or onto plastic tubes, but not in the more random, assay systems which utilised magnetic particles as the separation phase. The solid-phase, coated-tube chemiluminometric assay was comparable with ELISA for the measurement of TPOAb in human serum."],"dc:identifier":["oai:pure.atira.dk:studenttheses/2f7b4682-04e3-44f4-a56f-25760683e7ab","https://pure.southwales.ac.uk/en/studentTheses/2f7b4682-04e3-44f4-a56f-25760683e7ab"],"dc:identifier.uri":["https://pure.southwales.ac.uk/files/2104139/K._C._Thomas_2000_2059454.pdf"],"dc:language":["eng"],"dc:relation.isreferencedby":["https://pure.southwales.ac.uk/en/studentTheses/2f7b4682-04e3-44f4-a56f-25760683e7ab"],"dc:subject":["Biochemistry","Autoantibodies","Thyroid Peroxida","TPOAb","immunoradiometric assay","IRMA","enzymeimmunometric assay","Chemiluminescent Assay","acridinium ester (AE)"],"dc:title":["Development of a Chemiluminescent Assay for Autoantibodies to Thyroid Peroxidase"],"dc:type":["Thesis"],"dc:type.qualificationlevel":["Student thesis"],"dc:type.qualificationname":["Master's Thesis"]},"updated_at":"2026-07-24T04:38:59Z"}