title: Enzyme replacement therapy in Gaucher's disease: large-scale purification of glucocerebrosidase suitable for human administration.
abstract: Enzyme replacement therapy for the alleviation of Gaucher's disease has been impeded because of the difficulty in preparing large amounts of glucocerebrosidase, the enzyme that is deficient in patients with this disorder. A large-scale procedure for the purification of human placental glucocerebrosidase has been developed. The method uses cholate extraction, ammonium sulfate fractionation, acid precipitation, butanol extraction, and hydrophobic chromatography; the final enzyme preparation has a specific activity of more than 10(6) units/mg of protein with an overall recovery of 30%. In addition, the contamination of enzyme preparations, intended for human infusion, prepared by isolation procedures involving concanavalin A columns has been studied and is reported here.
mesh_terms: D005260:Female; D005776:Gaucher Disease / Q000188:drug therapy*; D005959:Glucosidases / Q000627:therapeutic use*; D005962:Glucosylceramidase / Q000302:isolation & purification / Q000627:therapeutic use*; D006801:Humans; D010920:Placenta / Q000201:enzymology; D011247:Pregnancy
chemical_list: D005959:Glucosidases; D005962:Glucosylceramidase
references.citation: Biochem Biophys Res Commun. 1965 Jan 18;18:221-5
references.pmid: 14282020
references.citation: Proc Natl Acad Sci U S A. 1976 Dec;73(12):4672-4
references.pmid: 1070017
references.citation: Biochem Biophys Res Commun. 1976 Aug 23;71(4):1048-53
references.pmid: 971299
references.citation: Proc Natl Acad Sci U S A. 1977 Mar;74(3):818-21
references.pmid: 322153
references.citation: J Neurochem. 1976 Dec;27(6):1513-9
references.pmid: 1003225
references.citation: Proc Soc Exp Biol Med. 1976 Nov;153(2):213-9
references.pmid: 995951
references.citation: J Exp Zool. 1976 Nov;198(2):261-6
references.pmid: 978175
references.citation: FEBS Lett. 1974 Aug 30;44(3):274-8
references.pmid: 4412570
references.citation: N Engl J Med. 1974 NOV 7;291(19):989-93
references.pmid: 4415565
references.citation: FEBS Lett. 1974 Jun 1;42(2):209-13
references.pmid: 4212005
references.citation: Int Rev Cytol. 1974;39:89-190
references.pmid: 4611947
references.citation: Proc Soc Exp Biol Med. 1974 Sep;146(4):1163-5
references.pmid: 4471637
references.citation: J Biol Chem. 1973 Aug 10;248(15):5256-61
references.pmid: 4768898
references.citation: J Biol Chem. 1973 May 10;248(9):3137-45
references.pmid: 4700453
references.citation: Anal Biochem. 1976 Jan;70(1):241-50
references.pmid: 1259145
references.citation: Nature. 1970 Aug 15;227(5259):680-5
references.pmid: 5432063
references.citation: J Biol Chem. 1966 Mar 10;241(5):1081-4
references.pmid: 5933867