Human α-N-Acetylglucosaminidase (Naglu) is a lysosomal acid hydrolase
implicated in tthe rare metabolic storage disorder known as mucopolysaccharidosis type
IIIB (MPS IIIB; also Sanfilippo syndrome B). Absence of this enzyme results in
cytotoxic accumulation of heparan sulphate in the central nervous system, causing mental
retardation and a shortened lifespan. Enzyme replacement therapy is not currently
effective to treat neurological symptoms due to the inability of exogenous Naglu to
access the brain. This laboratory uses a Spodoptera frugiperda (Sf9) insect cell system to
express Naglu fused to a synthetic protein transduction domain with the intent to
facilitate delivery of Naglu across the blood-brain barrier.
The project described herein may be broken down into three main sections.
Firstly, the impact of two cryptic splice sites on Naglu expression levels was analyzed in
both transiently expressing Sf9 cultures and stably selected cell lines. Secondly, the
effectiveness of the native Naglu secretion-signaling peptide in the Sf9 system was
examined. Finally, purification of a Naglu fusion protein from suspension culture
medium was performed using hydrophobic interaction chromatographic techniques.
The ultimate goal of this research is to develop an efficient system for
economical, large-scale production of a human recombinant Naglu fusion protein that has
the potential to be successfully used for enzyme replacement therapy to treat MPS IIIB. / Graduate
| Identifer | oai:union.ndltd.org:uvic.ca/oai:dspace.library.uvic.ca:1828/3455 |
| Date | 15 August 2011 |
| Creators | Jantzen, Roni Rebecca |
| Contributors | Choy, Francis Y. M. |
| Source Sets | University of Victoria |
| Language | English, English |
| Detected Language | English |
| Type | Thesis |
| Rights | Available to the World Wide Web |
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