Hicham Naimy1, Kendall D Powell2, John R Moriarity2, Jiang Wu1, Thomas G McCauley1, Patrick Aj Haslett1,3, Ann J Barbier1,4, Yongchang Qiu1. 1. Shire, 300 Shire Way, Lexington, MA 02421, USA. 2. Tandem Labs - RTP, A LabCorp Company, 2202 Ellis Road, Suite D, Durham, NC 27703, USA. 3. Current address: Agios Pharmaceuticals, 88 Sidney Street, Cambridge, MA 02139, USA. 4. Current address: Alnylam Pharmaceuticals, Inc, 300 Third Street, Cambridge, MA 02142, USA.
Abstract
AIMS: Heparan sulfate (HS) accumulates in the central nervous system in mucopolysaccharidosis III type A (MPS IIIA). A validated LC-MS/MS assay was developed to measure HS in human cerebrospinal fluid (CSF). METHODS & RESULTS: HS was extracted and digested and the resultant disaccharides were derivatized with a novel label, 4-butylaniline, enabling isoform separation and isotope-tagged analog introduction as an internal standard for LC-MS/MS. The assay has a LLOQ for disaccharides of 0.1 μM, ±20% accuracy and ≤20% precision. CSF samples from patients with MPS IIIA showed elevated HS levels (mean 4.9 μM) compared with negative controls (0.37 μM). CONCLUSION: This assay detected elevated HS levels in the CSF of patients with MPS IIIA and provides a method to assess experimental therapies.
AIMS: Heparan sulfate (HS) accumulates in the central nervous system in mucopolysaccharidosis III type A (MPS IIIA). A validated LC-MS/MS assay was developed to measure HS in human cerebrospinal fluid (CSF). METHODS & RESULTS:HS was extracted and digested and the resultant disaccharides were derivatized with a novel label, 4-butylaniline, enabling isoform separation and isotope-tagged analog introduction as an internal standard for LC-MS/MS. The assay has a LLOQ for disaccharides of 0.1 μM, ±20% accuracy and ≤20% precision. CSF samples from patients with MPS IIIA showed elevated HS levels (mean 4.9 μM) compared with negative controls (0.37 μM). CONCLUSION: This assay detected elevated HS levels in the CSF of patients with MPS IIIA and provides a method to assess experimental therapies.
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