{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T02:32:13Z","timestamp":1790821933682,"version":"4.1.0"},"reference-count":38,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2017,9,4]],"date-time":"2017-09-04T00:00:00Z","timestamp":1504483200000},"content-version":"vor","delay-in-days":3,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100002923","name":"CONICET","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002923","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005746","name":"Universidad Nacional del Litoral","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100005746","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotechnology Progress"],"published-print":{"date-parts":[[2017,9]]},"abstract":"<jats:p>\n                    Fabry disease is an X\u2010linked recessive disorder caused by a deficiency in lysosomal \u03b1\u2010Galactosidase A. Currently, two enzyme replacement therapies (ERT) are available. However, access to orphan drugs continues to be limited by their high price. Selection of adequate high\u2010expression systems still constitutes a challenge for alleviating the cost of treatments. Several strategies have been implemented, with varying success, trying to optimize the production process of recombinant human \u03b1\u2010Galactosidase A (rh\u03b1GAL) in Chinese hamster ovary (CHO\u2010K1) cells. Herein, we describe for the first time the application of a strategy based on third\u2010generation lentiviral particles (LP) transduction of suspension CHO\u2010K1 cells to obtain high\u2010producing rh\u03b1GAL clones (3.5 to 59.4 pg cell\n                    <jats:sup>\u22121<\/jats:sup>\n                    d\n                    <jats:sup>\u22121<\/jats:sup>\n                    ). After two purification steps, the active enzyme was recovered (2.4 \u00d7 10\n                    <jats:sup>6<\/jats:sup>\n                    U mg\n                    <jats:sup>\u22121<\/jats:sup>\n                    ) with 98% purity and 60% overall yield. Michaelis\u2010Menten analysis demonstrated that rh\u03b1GAL was capable of hydrolyzing the synthetic substrate 4MU\u2010\u03b1\u2010Gal at a comparable rate to Fabrazyme\u00ae, the current CHO\u2010derived ERT available for Fabry disease. In addition, rh\u03b1GAL presented the same mannose\u20106\u2010phosphate (M6P) content, about 40% higher acid sialic amount and 33% reduced content of the immunogenic type of sialic acid (Neu5Gc) than the corresponding ones for Fabrazyme\u00ae. In comparison with other rh\u03b1GAL production processes reported to date, our approach achieves the highest rh\u03b1GAL productivity preserving adequate activity and glycosylation pattern. Even more, considering the improved glycosylation characteristics of rh\u03b1GAL, which might provide advantages regarding pharmacokinetics, our enzyme could be postulated as a promising alternative for therapeutic use in Fabry disease. \u00a9 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1334\u20131345, 2017\n                  <\/jats:p>","DOI":"10.1002\/btpr.2538","type":"journal-article","created":{"date-parts":[[2017,8,25]],"date-time":"2017-08-25T04:22:47Z","timestamp":1503634967000},"page":"1334-1345","source":"Crossref","is-referenced-by-count":4,"title":["High yield process for the production of active human \u03b1\u2010galactosidase a in CHO\u2010K1 cells through lentivirus transgenesis"],"prefix":"10.1002","volume":"33","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7640-6221","authenticated-orcid":false,"given":"Mar\u00eda Celeste","family":"Rodr\u00edguez","sequence":"first","affiliation":[{"name":"Universidad Nacional del Litoral, CONICET, School of Biochemistry and Biological Sciences, Cell Culture Laboratory, Ciudad Universitaria, Paraje El Pozo, C.C. 242 Santa Fe S3000ZAA Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natalia","family":"Ceaglio","sequence":"additional","affiliation":[{"name":"Universidad Nacional del Litoral, CONICET, School of Biochemistry and Biological Sciences, Cell Culture Laboratory, Ciudad Universitaria, Paraje El Pozo, C.C. 242 Santa Fe S3000ZAA Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sebasti\u00e1n","family":"Antu\u00f1a","sequence":"additional","affiliation":[{"name":"Zelltek S.A., PTLC RN 168 Santa Fe S3000ZAA Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mar\u00eda Bel\u00e9n","family":"Tardivo","sequence":"additional","affiliation":[{"name":"Zelltek S.A., PTLC RN 168 Santa Fe S3000ZAA Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marina","family":"Etcheverrigaray","sequence":"additional","affiliation":[{"name":"Universidad Nacional del Litoral, CONICET, School of Biochemistry and Biological Sciences, Cell Culture Laboratory, Ciudad Universitaria, Paraje El Pozo, C.C. 242 Santa Fe S3000ZAA Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudio","family":"Prieto","sequence":"additional","affiliation":[{"name":"Universidad Nacional del Litoral, School of Biochemistry and Biological Sciences, Cell Culture Laboratory, Ciudad Universitaria, Paraje El Pozo, C.C. 242 Santa Fe S3000ZAA Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2017,9,4]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymgme.2014.08.002"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrd3275"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymgme.2012.08.003"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/cwg034"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10038-005-0342-9"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1002\/btpr.637"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1186\/1753-6561-5-S8-P7"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.22265"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2013.11.035"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1002\/btpr.2447"},{"key":"e_1_2_8_12_1","unstructured":"PlewaC.Application of lentiviral vectors for development of production cell lines and safety testing of lentiviral\u2010derived cells or products application of lentiviral vectors for development of production cell lines and safety testing of lentiviral\u2010derived cells;2010."},{"key":"e_1_2_8_13_1","unstructured":"SohnY LeeJM ParkH\u2010R JungS\u2010C ParkTH OhD\u2010B.Enhanced sialylation and in vivo efficacy of recombinant human \u03b1\u2010galactosidase through in vitro glycosylation.2013;6696:0\u20135."},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pep.2013.11.005"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.72.11.8463-8471.1998"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.72.10.8150-8157.1998"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1038\/mt.2010.239"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1021\/bp049825t"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1002\/jctb.940"},{"key":"e_1_2_8_20_1","unstructured":"Richard FSelden MarianneBorowski Carol MKinoshita Douglas ATreco Melanie DWilliams Thomas JSchuetz PFD. 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