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Claims for Patent: 5,236,838

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Claims for Patent: 5,236,838

Title: Enzymatically active recombinant glucocerebrosidase
Abstract:Recombinant enzymatically active glucocerebrosidase is produced by a eukaryotic cell. Also, a cell includes nucleic acid encoding enzymatically active glucocerebrosidase; also a eukaryotic organism contains such a cell. Also, a method for producing enzymatically active glucocerebrosidase includes steps of introducing glucocerebrosidase-encoding nucleic acid into a eukaryotic cell, causing the cell to express glucocerebrosidase, and purifying the glucocerebrosidase from the cell.
Inventor(s): Rasmussen; James (Boston, MA), Barsomian; Gary (Georgetown, MA), Bergh; Michel (Belmont, MA)
Assignee: Genzyme Corporation (Cambridge, MA)
Application Number:07/748,283
Patent Claims: 1. A method for producing enzymatically active glucocerebrosidase comprising the steps of:

introducing nucleic acid encoding human glucocerebrosidase into a CHO cell;

causing said cell to express and secrete said glucocerebrosidase into a culture medium; and

purifying said glucocerebrosidase from said culture medium.

2. The method of claim 1 wherein the pH of said culture medium is between about pH 6.5 and pH 7.2.

3. The method of claim 2 wherein the pH of said culture medium is between about pH 6.6 and pH 6.8.

4. The method of claim 1 wherein said culture medium contains O.sub.2 in an amount below about 50% saturation and sufficient to maintain the cells.

5. The method of claim 1 wherein said culture medium contains O.sub.2 in an amount between about 20% saturation and about 30% saturation.

6. A method for producing enzymatically active glucocerebrosidase comprising the steps of:

introducing a plasmid containing nucleic acid encoding glucocerebrosidase into a CHO cell, said plasmid being selected from the group consisting of pGB20, pGB37 and pGB42;

causing said cell to express and secrete said glucocerebrosidase into a culture medium; and

purifying said glucocerebrosidase from said culture medium.
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