Carroll, Kenneth (1988) Monoclonal antibody production and the application of monoclonal antibodies to the study of tumour cell membrane antigens. PhD thesis, Dublin City University.
Abstract
A murine monoclonal antibody, F10 1 E12, (IgM) was
produced against Landschutz ascites tumour cells (LAT)
This antibody exhibits preferential reactivity on tumour
versus normal cells The antigen against which the
antibody is directed was identified as a cell membrane
glycoprotein of mw 205kD, designated p-205
Characterization of this antigen by Western immunoblot
analysis, coupled with enzyme digestion studies, revealed
that the epitope bound by F10 1 E12 is sensitive to the
effects of trypsin, pronase and papain. This suggests
that an arginine and/or lysine ammoacid residue may be
located at this site Lipase and a number of
glycosidases did not affect the antigen-antibody
interaction Neuraminidase was found to destroy this
interaction, however, it did lead to the immunodetection
of a range of other glycoproteins of mw from 40kD to
60kD. This may have been due to the effects of undefined
proteases in the neuraminidase preparation used.
The p-205 of LAT cells was not detectable on a range of
normal mouse cells by Western immunoblot analysis,
despite the fact that certain of these cells, notably
normal mouse liver cells, exhibited strong reaction with
F10 1 E12 m solid-phase ELISA studies. This would
suggest that the antigen detected on normal cells is
different to that of LAT cells, or that the antigen as
expressed by normal cells is more labile than that of
tumour cells.
The glycoprotein p-205 was also detectable by Western
immunoblotting analysis using reagent polyclonal
antibodies raised against nigh molecular weight
fucopeptides derived from LAT cell surfaces. These
fucopeptides are well characterized m the literature and
demonstrate a strong association with the neoplastic
state. This finding may provide some valuable clues as
to the nature of the carbohydrate moieties of p-205.
In the course of this project, developments were made m
the areas of solid-phase ELISA, somatic cell fusion
techniques using PEG and m the elimination of mycoplasma
from infected hybridomas. These methods and detailed
accounts of the developments initiated during this
research assignment will also be discussed.
Metadata
Item Type: | Thesis (PhD) |
---|---|
Date of Award: | 1988 |
Refereed: | No |
Supervisor(s): | O'Kennedy, Richard |
Uncontrolled Keywords: | Monoclonal antibodies; Antigen-antibody reactions |
Subjects: | Biological Sciences > Biotechnology |
DCU Faculties and Centres: | DCU Faculties and Schools > Faculty of Science and Health > School of Biotechnology |
Use License: | This item is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 3.0 License. View License |
ID Code: | 18401 |
Deposited On: | 17 Jul 2013 13:04 by Celine Campbell . Last Modified 17 Jul 2013 13:04 |
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