Study of a monoclonal antibody to human B cells
Thesis (M.Sc.)--Memorial University of Newfoundland, 1986. Medicine Bibliography: leaves 137-163. The purpose of this work was to characterize and determine the specificity of a mouse monoclonal antibody (NFLD.M1) which was derived from a fusion between SP2/0-Ag14 and spleen cells from a Balb/c mous...
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ftmemorialunivdc:oai:collections.mun.ca:theses2/65076 2023-05-15T17:23:30+02:00 Study of a monoclonal antibody to human B cells Drover, Mary Sheila Lewis Memorial University of Newfoundland. Faculty of Medicine 1986 xviii, 167 leaves : ill. Image/jpeg; Application/pdf http://collections.mun.ca/cdm/ref/collection/theses2/id/65076 Eng eng Electronic Theses and Dissertations (22.37 MB) -- http://collections.mun.ca/PDFs/theses/Drover_MarySheilaLewis.pdf 75380783 http://collections.mun.ca/cdm/ref/collection/theses2/id/65076 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 Monoclonal antibodies HLA histocompatibility antigens Text Electronic thesis or dissertation 1986 ftmemorialunivdc 2015-08-06T19:16:40Z Thesis (M.Sc.)--Memorial University of Newfoundland, 1986. Medicine Bibliography: leaves 137-163. The purpose of this work was to characterize and determine the specificity of a mouse monoclonal antibody (NFLD.M1) which was derived from a fusion between SP2/0-Ag14 and spleen cells from a Balb/c mouse that had been hyperimmunized with B-cells from a chronic lymphatic leukemic patient. Cloning was done by limiting dilution and positive clones were selected by screening on a panel of viable cells using the cellular enzyme-linked immunosorbent assay (CELISA). This assay was shown to be more specific and sensitive than an ELISA that used glutaraldehyde-fixed cells. -- Two sources of the antibody (purified IgG1 from ascites fluid and supernatant from overgrown cultures) appeared to be identical in their serological pattern on several B-cell lines. Specificity testing using the CELISA and several different cell types revealed that NFLD.M1 recognized some B-cells, but failed to react with any of the T-cells tested. A Frequency Distribution plot of the data showed that NFLD.M1 reacted with the cells in a bimodal fashion compared to the normal distribution observed with the monomorphic monoclonal antibody, NEI anti-Ia. Furthermore when NFLD.M1 antibody was expressed as a percent of the NEI anti-Ia it was found that all the DR4 positive cells produced values greater than 50% whereas DR4 negative cells gave values less than 30%. Using 30% as a cutoff point a correlation analysis was done on the CELISA results for 42 cell lines. The r value obtained for DR4 and NFLD.M1 was 1 with a p value of 2 x 10-10. In addition significant r values were obtained for DRw53 and DQw3 which are in linkage disequilibrium with DR4. -- 0ne-dimensional electrophoresis of the immunoprecipitated molecules from a DR4 cell produced a banding pattern that was compatible with that of the alpha and beta subunits of DR. Thesis Newfoundland studies University of Newfoundland Memorial University of Newfoundland: Digital Archives Initiative (DAI) |
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Memorial University of Newfoundland: Digital Archives Initiative (DAI) |
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English |
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Monoclonal antibodies HLA histocompatibility antigens |
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Monoclonal antibodies HLA histocompatibility antigens Drover, Mary Sheila Lewis Study of a monoclonal antibody to human B cells |
topic_facet |
Monoclonal antibodies HLA histocompatibility antigens |
description |
Thesis (M.Sc.)--Memorial University of Newfoundland, 1986. Medicine Bibliography: leaves 137-163. The purpose of this work was to characterize and determine the specificity of a mouse monoclonal antibody (NFLD.M1) which was derived from a fusion between SP2/0-Ag14 and spleen cells from a Balb/c mouse that had been hyperimmunized with B-cells from a chronic lymphatic leukemic patient. Cloning was done by limiting dilution and positive clones were selected by screening on a panel of viable cells using the cellular enzyme-linked immunosorbent assay (CELISA). This assay was shown to be more specific and sensitive than an ELISA that used glutaraldehyde-fixed cells. -- Two sources of the antibody (purified IgG1 from ascites fluid and supernatant from overgrown cultures) appeared to be identical in their serological pattern on several B-cell lines. Specificity testing using the CELISA and several different cell types revealed that NFLD.M1 recognized some B-cells, but failed to react with any of the T-cells tested. A Frequency Distribution plot of the data showed that NFLD.M1 reacted with the cells in a bimodal fashion compared to the normal distribution observed with the monomorphic monoclonal antibody, NEI anti-Ia. Furthermore when NFLD.M1 antibody was expressed as a percent of the NEI anti-Ia it was found that all the DR4 positive cells produced values greater than 50% whereas DR4 negative cells gave values less than 30%. Using 30% as a cutoff point a correlation analysis was done on the CELISA results for 42 cell lines. The r value obtained for DR4 and NFLD.M1 was 1 with a p value of 2 x 10-10. In addition significant r values were obtained for DRw53 and DQw3 which are in linkage disequilibrium with DR4. -- 0ne-dimensional electrophoresis of the immunoprecipitated molecules from a DR4 cell produced a banding pattern that was compatible with that of the alpha and beta subunits of DR. |
author2 |
Memorial University of Newfoundland. Faculty of Medicine |
format |
Thesis |
author |
Drover, Mary Sheila Lewis |
author_facet |
Drover, Mary Sheila Lewis |
author_sort |
Drover, Mary Sheila Lewis |
title |
Study of a monoclonal antibody to human B cells |
title_short |
Study of a monoclonal antibody to human B cells |
title_full |
Study of a monoclonal antibody to human B cells |
title_fullStr |
Study of a monoclonal antibody to human B cells |
title_full_unstemmed |
Study of a monoclonal antibody to human B cells |
title_sort |
study of a monoclonal antibody to human b cells |
publishDate |
1986 |
url |
http://collections.mun.ca/cdm/ref/collection/theses2/id/65076 |
genre |
Newfoundland studies University of Newfoundland |
genre_facet |
Newfoundland studies University of Newfoundland |
op_source |
Paper copy kept in the Centre for Newfoundland Studies, Memorial University Libraries |
op_relation |
Electronic Theses and Dissertations (22.37 MB) -- http://collections.mun.ca/PDFs/theses/Drover_MarySheilaLewis.pdf 75380783 http://collections.mun.ca/cdm/ref/collection/theses2/id/65076 |
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_ |
1766112832959545344 |