Date of Award

1998

Document Type

Master Thesis

Degree Name

Masters of Science (Research)

Department

Biological Sciences

First Advisor

Dr. J. O'Mullane

Second Advisor

Dr. M. Madden

Third Advisor

Dr. S. Fanning

Abstract

CD44 is a cell surface glycoprotein (mol. wt. =85-95 kDa) which has been implicated in lymphocyte-HEV interactions and the process of lymphocyte homing. CD44 is a complex glycoprotein which undergoes extensive alternative splicing to generate various CD44 isoforms. CD44 functions include the delivery of an activation signal to T-cells, binding to hyaluronate, mediation of lymphocyte homing, and the spread of metastatic cells. CD44 has been particularly implicated in the dissemination of non- Hodgkin's lymphoma and as such may serve as a useful prognostic marker of disease progression. CD44 measurement in serum may provide a rapid, inexpensive and relatively non-invasive prognostic marker for NHL. Commercial ELISA systems have been developed for such a purpose and were assessed as part of this study. Two assays were evaluated but both were found to be unsatisfactory. The lack of efficacy of the assays led to the development of an 'in-house' ELISA system. The development of such a system requires CD44 antigen, which despite several approaches to the various manufacturers, proved impossible to obtain. Cloned CD44 cDNA was therefore used to generate antigen by recombinant DNA methods. CD44 cDNA was subcloned into the pTrcHis2 expression vector . CD44 protein expression was induced by IPTG and purified by affinity chromatography. The presence of the protein was confirmed by SDS-PAGE and western blot analysis. The protein was quantified using a Hitachi 911 analyser.

Comments

A thesis presented to the National Council for Educational Awards for the degree of Master of Science

Access Level

info:eu-repo/semantics/openAccess

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