{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T18:11:52Z","timestamp":1760551912428,"version":"build-2065373602"},"reference-count":16,"publisher":"Wiley","issue":"15","license":[{"start":{"date-parts":[[2005,4,14]],"date-time":"2005-04-14T00:00:00Z","timestamp":1113436800000},"content-version":"vor","delay-in-days":2356,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Electrophoresis"],"published-print":{"date-parts":[[1998,11]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The lysosomal storage disorder mucopolysaccharidoses I (MPS I) is caused by a deficiency in the production of \u03b1\u2010<jats:sc>L<\/jats:sc>\u2010iduronidase. Recently, a recombinant \u03b1\u2010<jats:sc>L<\/jats:sc>\u2010iduronidase has been produced in Chinese hamster ovary (CHO) cells. It is thought that for \u03b1\u2010<jats:sc>L<\/jats:sc>\u2010iduronidase to be correctly targeted to the lysosomal vesicle a particular oligosaccharide make\u2010up must be present, and characterization of the carbohydrates is critical. Oligosaccharides from \u03b1\u2010<jats:sc>L<\/jats:sc>\u2010iduronidase were analyzed using fluorophore\u2010assisted carbohydrate electrophoresis (FACE\u00ae). The FACE\u00ae system uses polyacrylamide gel electrophoresis to separate, quantify, and determine the sequence of oligosaccharides released from glycoproteins. Asparagine\u2010linked oligosaccharides were released from \u03b1\u2010<jats:sc>L<\/jats:sc>\u2010iduronidase using the enzyme peptide <jats:italic>N<\/jats:italic>\u2010glycanase F (PNGase F). Released oligosaccharides were labeled with a fluorophore at the reducing termini by reductive amination. A total of nine bands were sequenced from the released pool of oligosaccharides. The pool of fluorescently labeled oligosaccharides was then electrophoresed in preparative gels and each band individually excised and extracted. Isolated bands were treated with a series of exoenzymes to determine the sequence of monosaccharides that make up a particular oligosaccharide. A total of eighteen different oligosaccharides were identified from the original pool of oligosaccharides. A majority of the oligosaccharides, over 73%, were found to be of the sialylated complex type. Four of the oligosaccharides were phosphorylated, making up approximately 11% of the carbohydrate pool, and the remaining 15% were of the oligomannose type.<\/jats:p>","DOI":"10.1002\/elps.1150191508","type":"journal-article","created":{"date-parts":[[2005,5,28]],"date-time":"2005-05-28T12:36:50Z","timestamp":1117283810000},"page":"2612-2620","source":"Crossref","is-referenced-by-count":9,"title":["Structural determination of oligosaccharides from recombinant iduronidase released with peptide <i>N<\/i>\u2010glycanase F using fluorophore\u2010assisted carbohydrate electrophoresis"],"prefix":"10.1002","volume":"19","author":[{"given":"Chuck","family":"Hague","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R. 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