Fluorimetric assay with a novel substrate for quantification of galactocerebrosidase activity in dried blood spot specimens.

ABSTRACT:

Background

Decreased galactocerebrosidase (GALC) enzyme activity is causative for Krabbe disease, a lysosomal storage disorder with devastating neurodegenerative consequences. Quantitative fluorimetric assays for GALC activity in isolated blood and skin cells have been described; however, no such assay has been described using dried blood spot (DBS) specimens.

Methods

GALC enzyme activity was measured quantitatively using fluorescence from a novel glycosidic substrate: carboxy derived from 6-hexadecanoylamino-4-methylumbelliferone. GALC activity was demonstrated on newborn DBS specimens, known Krabbe disease patient specimens, proficiency testing and quality control samples.

Results

We present data on characterization of the novel substrate and assay, including pH opt

SUBMITTER: Ullal AJ 

PROVIDER: S-EPMC6838535 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature