title: Glycosylation and functionality of recombinant β-glucocerebrosidase from various production systems.
abstract: The glycosylation of recombinant β-glucocerebrosidase, and in particular the exposure of mannose residues, has been shown to be a key factor in the success of ERT (enzyme replacement therapy) for the treatment of GD (Gaucher disease). Macrophages, the target cells in GD, internalize β-glucocerebrosidase through MRs (mannose receptors). Three enzymes are commercially available for the treatment of GD by ERT. Taliglucerase alfa, imiglucerase and velaglucerase alfa are each produced in different cell systems and undergo various post-translational or post-production glycosylation modifications to expose their mannose residues. This is the first study in which the glycosylation profiles of the three enzymes are compared, using the same methodology and the effect on functionality and cellular uptake is evaluated. While the major differences in glycosylation profiles reside in the variation of terminal residues and mannose chain length, the enzymatic activity and stability are not affected by these differences. Furthermore, the cellular uptake and in-cell stability in rat and human macrophages are similar. Finally, in vivo studies to evaluate the uptake into target organs also show similar results for all three enzymes. These results indicate that the variations of glycosylation between the three regulatory-approved β-glucocerebrosidase enzymes have no effect on their function or distribution.
authors: *Protalix Biotherapeutics, 2 Snunit Street, P. O. Box 455, Carmiel, 2161401, Israel.
authors: Yoram
authors: Y
authors: Tekoah
authors: *Protalix Biotherapeutics, 2 Snunit Street, P. O. Box 455, Carmiel, 2161401, Israel.
authors: Salit
authors: S
authors: Tzaban
authors: *Protalix Biotherapeutics, 2 Snunit Street, P. O. Box 455, Carmiel, 2161401, Israel.
authors: Tali
authors: T
authors: Kizhner
authors: *Protalix Biotherapeutics, 2 Snunit Street, P. O. Box 455, Carmiel, 2161401, Israel.
authors: Mariana
authors: M
authors: Hainrichson
authors: *Protalix Biotherapeutics, 2 Snunit Street, P. O. Box 455, Carmiel, 2161401, Israel.
authors: Anna
authors: A
authors: Gantman
authors: *Protalix Biotherapeutics, 2 Snunit Street, P. O. Box 455, Carmiel, 2161401, Israel.
authors: Myriam
authors: M
authors: Golembo
authors: David
authors: D
authors: Aviezer
authors: *Protalix Biotherapeutics, 2 Snunit Street, P. O. Box 455, Carmiel, 2161401, Israel.
authors: Yoseph
authors: Y
authors: Shaaltiel
mesh_terms: D000818:Animals; D001692:Biological Transport; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002460:Cell Line; D004795:Enzyme Stability; D005962:Glucosylceramidase / Q000737:chemistry / Q000378:metabolism* / Q000493:pharmacokinetics; D006031:Glycosylation; D006801:Humans; D007700:Kinetics; D016676:Macrophages, Alveolar / Q000201:enzymology; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D014018:Tissue Distribution
chemical_list: D011994:Recombinant Proteins; D005962:Glucosylceramidase; C550184:Velaglucerase alfa, human; C582473:taliglucerase alfa; C090568:imiglucerase
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