PMID: 19864504

 

    Legend: Gene, Sites

Title : Site-specific analysis of N-linked oligosaccharides of recombinant lysosomal arylsulfatase A produced in different cell lines

Abstract :

1. Metachromatic leukodystrophy ( MLD ) is a lysosomal storage disease caused by a deficiency of the lysosomal enzyme arylsulfatase A ( ASA )
2. Enzyme replacement therapy ( ERT ) is a therapeutic option for MLD and other lysosomal disorders
3. This therapy depends on N-linked oligosaccharide-mediated delivery of intravenously injected recombinant enzyme to the lysosomes of patient cells
4. Because of the importance of N-linked oligosaccharide side chains in ERT , we examined the com position of the three N-linked glycans of four different recombinant ASAs in a site-specific manner
5. Depending on the culture conditions and the cell line expressing the enzyme , we detected a high variability of the high-mannose-type N-glycans which prevail at all glycosylation sites
6. Our data show that the com position of the glycans is largely determined by substantial trimming in the medium
7. The susceptibility for trimming is different for the glycans at the three N-glycosylation sites
8. Interestingly, which of the glycans is most susceptible to trimming also depends on production conditions
9. CHO cells cultured under bioreactor conditions yielded recombinant ASA with the most preserved N-glycan structures, the highest mannose-6-phosphate content and the highest similarity to non-recombinant enzyme
10. Notably, roughly one-third of the N-glycans released from the three glycosylation sites were fucosylated
11. In the last years, numerous recombinant lysosomal enzymes were used for preclinical ERT trials
12. Our data show that the oligosaccharide structures were very different in these trials making it difficult to draw common conclusions from the various investigations

Output (sent_index, trigger, protein, sugar, site):

• 0. A, , arylsulfatase A, N-linked oligosaccharides, -
• 10. fucosylated, , -, the N-glycans, -
• 10. glycosylation, , -, -, sites
• 10. released, , -, the N-glycans, sites
• 3. enzyme, , enzyme, N-linked oligosaccharide-mediated delivery, -
• 7. N-glycosylation, , -, -, sites
• 7. sites, , -, the glycans, sites
• 9. ASA, , ASA, the highest mannose-6-phosphate content, -
• 9. ASA, , ASA, the most preserved N-glycan structures, -

Output(Part-Of) (sent_index, protein, site):

*Output_Site_Fusion* (sent_index, protein, sugar, site):

 

Protein	NCBI ID	SENTENCE INDEX
arylsulfatase A	410	0