Study of a monoclonal antibody to human B cells

Drover, Mary Sheila Lewis (1986) Study of a monoclonal antibody to human B cells. Masters thesis, Memorial University of Newfoundland.

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    Available under License - 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.
    (Original Version)

Abstract

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. -- One-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.

Item Type: Thesis (Masters)
URI: http://research.library.mun.ca/id/eprint/5611
Item ID: 5611
Additional Information: Bibliography: leaves 137-163.
Department(s): Medicine, Faculty of
Date: 1986
Date Type: Submission
Library of Congress Subject Heading: Monoclonal antibodies; HLA histocompatibility antigens
Medical Subject Heading: Antibodies, Monoclonal; B-Lymphocytes; HLA Antigens

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