A Biomarker for the Degradation of Glycogen by Recombinant Lysosomal α-Glucosidase (rhGAA) in vitro and It’s Presence in Serum In vivo


Pre-Clinical Research

Poster Number: Virtual


Allen Murray, Ph.D., Glycan Technologies, Inc.

In studies on the degradation of glycogen by rhGAA, a glycosylated protein residual material was found which consists of about 5–6% of the total starting glycogen. There was an additional 25% of the glycogen unaccounted for based on glucose released. After incubation of glycogen with rhGAA until no more glucose was released, no other carbohydrate was detected on HPAEC-PAD. Several oligosaccharides are then detectable if the medium is first boiled in 0.1NHCl or incubated with a protease. It is present in serum either in an HCl extract or in a trypsin digest. The characteristics of the in vivo serum material are identical to the material in the in vitro incubation medium. One oligosaccharide which cannot be further degraded by rhGAA, from the incubation medium as well as from serum co-elute on HPAEC-PAD. Several masked oligosaccharides in serum contain m-inositol, e-inositol, and sorbitol as the major carbohydrates. The presence of this glycosylated protein in serum is a fraction of glycogen that is degraded outside the lysosome and the cell. The glycosylated protein in the serum is not present in the serum of Pompe mice not on ERT, but it is present in the serum of Pompe disease patients who are on ERT, it is a biomarker of GAA degradation of lysosomal glycogen. Levels of the biomarker will be determined to monitor enzyme replacement therapy in patients. Since there are reports that the lysosomal degradation is on a diurnal rhythm this will be investigated. If there is a diurnal rhythm that will be accounted for in monitoring ERT.