{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,14]],"date-time":"2026-05-14T14:23:01Z","timestamp":1778768581772,"version":"3.51.4"},"reference-count":67,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2001,3,2]],"date-time":"2001-03-02T00:00:00Z","timestamp":983491200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotech &amp; Bioengineering"],"published-print":{"date-parts":[[2001,5,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Chinese hamster ovary and murine myeloma NS0 cells are currently favored host cell types for the production of therapeutic recombinant proteins. In this study, we compared <jats:italic>N<\/jats:italic>\u2010glycan processing in GS\u2010NS0 and GS\u2010CHO cells producing the same model recombinant glycoprotein, tissue inhibitor of metalloproteinases 1. By manipulation of intracellular nucleotide\u2010sugar content, we examined the feasibility of implementing metabolic control strategies aimed at reducing the occurrence of murine\u2010specific glycan motifs on NS0\u2010derived recombinant proteins, such as Gal\u03b11,3Gal\u03b21,4GlcNAc. Although both CHO and NS0\u2010derived oligosaccharides were predominantly of the standard complex type with variable sialylation, 30% of <jats:italic>N<\/jats:italic>\u2010glycan antennae associated with NS0\u2010derived TIMP\u20101 terminated in \u03b11,3\u2010linked galactose residues. Furthermore, NS0 cells conferred a greater proportion of terminal <jats:italic>N<\/jats:italic>\u2010glycolylneuraminic (sialic) acid residues as compared with the <jats:italic>N<\/jats:italic>\u2010acetylneuraminic acid variant. Inclusion of the nucleotide\u2010sugar precursors, glucosamine (10 m<jats:italic>M<\/jats:italic>, plus 2 m<jats:italic>M<\/jats:italic> uridine) and <jats:italic>N<\/jats:italic>\u2010acetylmannosamine (20 m<jats:italic>M<\/jats:italic>), in culture media were shown to significantly increase the intracellular pools of UDP\u2010<jats:italic>N<\/jats:italic>\u2010acetylhexosamine and CMP\u2010sialic acid, respectively, in both NS0 and CHO cells. The elevated UDP\u2010<jats:italic>N<\/jats:italic>\u2010acetylhexosamine content induced by the glucosamine\/uridine treatment was associated with an increase in the antennarity of <jats:italic>N<\/jats:italic>\u2010glycans associated with TIMP\u20101 produced in CHO cells but not <jats:italic>N<\/jats:italic>\u2010glycans associated with TIMP\u20101 from NS0 cells. In addition, elevated UDP\u2010<jats:italic>N<\/jats:italic>\u2010acetylhexosamine content was associated with a slight decrease in sialylation in both cell lines. The elevated CMP\u2010sialic acid content induced by <jats:italic>N<\/jats:italic>\u2010acetylmannosamine had no effect on the overall level of sialylation of TIMP\u20101 produced by both CHO and NS0 cells, although the ratio of <jats:italic>N<\/jats:italic>\u2010glycolylneuraminic acid:<jats:italic>N<\/jats:italic>\u2010acetylneuraminic acid associated with NS0\u2010derived TIMP\u20101 changed from 1:1 to 1:2. These data suggest that manipulation of nucleotide\u2010sugar metabolism can promote changes in <jats:italic>N<\/jats:italic>\u2010glycan processing that are either conserved between NS0 and CHO cells or specific to either NS0 cells or CHO cells.  \u00a9 2001 John Wiley &amp; Sons, Inc. Biotechnol Bioeng 73: 188\u2013202, 2001<\/jats:p>","DOI":"10.1002\/bit.1051","type":"journal-article","created":{"date-parts":[[2002,8,25]],"date-time":"2002-08-25T17:44:55Z","timestamp":1030297495000},"page":"188-202","source":"Crossref","is-referenced-by-count":157,"title":["Metabolic control of recombinant protein <i>N<\/i>\u2010glycan processing in NS0 and CHO cells"],"prefix":"10.1002","volume":"73","author":[{"given":"Kym N.","family":"Baker","sequence":"first","affiliation":[]},{"given":"Mark H.","family":"Rendall","sequence":"additional","affiliation":[]},{"given":"Anna E.","family":"Hills","sequence":"additional","affiliation":[]},{"given":"Michael","family":"Hoare","sequence":"additional","affiliation":[]},{"given":"Robert B.","family":"Freedman","sequence":"additional","affiliation":[]},{"given":"David C.","family":"James","sequence":"additional","affiliation":[]}],"member":"311","published-online":{"date-parts":[[2001,3,2]]},"reference":[{"key":"e_1_2_1_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.260470112"},{"key":"e_1_2_1_3_1","first-page":"69","article-title":"Real\u2010time monitoring of recombinant protein concentration in animal cell cultures using an optical biosensor","volume":"17","author":"Baker K","year":"1997","journal-title":"Gen Eng Biotechnol"},{"key":"e_1_2_1_4_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.19.12616"},{"key":"e_1_2_1_5_1","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1006\/abio.1995.1468","article-title":"Nonselective and efficient fluorescent labeling of glycans using 2\u2010aminobenzamide and anthranilic acid","volume":"230","author":"Bigge JC","year":"1995","journal-title":"Anal Biochem"},{"key":"e_1_2_1_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/0167-5699(93)90259-N"},{"key":"e_1_2_1_7_1","doi-asserted-by":"publisher","DOI":"10.1042\/bj2240277"},{"key":"e_1_2_1_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(80)80036-5"},{"key":"e_1_2_1_9_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.21.13622"},{"key":"e_1_2_1_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt0790-662"},{"key":"e_1_2_1_11_1","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/10.1.11"},{"key":"e_1_2_1_12_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1997.00187.x"},{"key":"e_1_2_1_13_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.93.15.7572"},{"key":"e_1_2_1_14_1","doi-asserted-by":"crossref","first-page":"2485","DOI":"10.1182\/blood.V82.8.2485.2485","article-title":"One percent of human girculating B\u2010lymphocytes are capable of producing the natural anti\u2010gal antibody","volume":"82","author":"Galili U","year":"1993","journal-title":"Blood"},{"key":"e_1_2_1_15_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0290(19980305)57:5<518::AID-BIT3>3.0.CO;2-J"},{"key":"e_1_2_1_16_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1997.t01-1-00187.x"},{"key":"e_1_2_1_17_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0290(19981205)60:5<596::AID-BIT10>3.0.CO;2-5"},{"key":"e_1_2_1_18_1","doi-asserted-by":"publisher","DOI":"10.1038\/37995"},{"key":"e_1_2_1_19_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1995.0718a.x"},{"key":"e_1_2_1_20_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1026466408042"},{"key":"e_1_2_1_21_1","doi-asserted-by":"publisher","DOI":"10.1021\/bp980005o"},{"key":"e_1_2_1_22_1","first-page":"191","volume-title":"New developments and new applications in animal cell technology: Proceedings of the 15th ESACT meeting","author":"Gregoire A","year":"1998"},{"key":"e_1_2_1_23_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0290(19980620)58:6<642::AID-BIT10>3.0.CO;2-9"},{"key":"e_1_2_1_24_1","doi-asserted-by":"publisher","DOI":"10.1006\/abio.1996.0351"},{"key":"e_1_2_1_25_1","doi-asserted-by":"crossref","first-page":"9005","DOI":"10.1016\/S0021-9258(19)50380-5","article-title":"Glycosylation of nuclear and cytoplasmic proteins \u2014 purification and characterization of a uridine diphospho\u2010N\u2010acetylglucosamine\u2010polypeptide beta\u2010N\u2010acetylglucosaminyltransferase","volume":"267","author":"Haltiwanger RS","year":"1992","journal-title":"J Biol Chem"},{"key":"e_1_2_1_26_1","doi-asserted-by":"publisher","DOI":"10.1093\/oxfordjournals.jbchem.a122838"},{"key":"e_1_2_1_27_1","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/6.7.711"},{"key":"e_1_2_1_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-2952(99)00035-0"},{"key":"e_1_2_1_29_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.39.24313"},{"key":"e_1_2_1_30_1","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/7.2.169"},{"key":"e_1_2_1_31_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1995.0981m.x"},{"key":"e_1_2_1_32_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0290(19990605)63:5<559::AID-BIT6>3.0.CO;2-L"},{"key":"e_1_2_1_33_1","doi-asserted-by":"publisher","DOI":"10.1006\/bbrc.1998.8946"},{"key":"e_1_2_1_34_1","doi-asserted-by":"publisher","DOI":"10.1093\/oxfordjournals.jbchem.a022076"},{"key":"e_1_2_1_35_1","first-page":"1336","volume-title":"Encyclopedia of bioprocess technology: Fermentation, biocatalysis and bioseparation","author":"James DC","year":"1999"},{"key":"e_1_2_1_36_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt0695-592"},{"key":"e_1_2_1_37_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-0348-7388-8_6"},{"key":"e_1_2_1_38_1","doi-asserted-by":"publisher","DOI":"10.1159\/000425635"},{"key":"e_1_2_1_39_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt0896-975"},{"key":"e_1_2_1_40_1","doi-asserted-by":"publisher","DOI":"10.1093\/oxfordjournals.jbchem.a022004"},{"key":"e_1_2_1_41_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.bi.39.070170.001451"},{"key":"e_1_2_1_42_1","doi-asserted-by":"publisher","DOI":"10.1042\/bj2170701"},{"key":"e_1_2_1_43_1","doi-asserted-by":"publisher","DOI":"10.1097\/00007890-199510270-00014"},{"key":"e_1_2_1_44_1","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/5.8.813"},{"key":"e_1_2_1_45_1","doi-asserted-by":"publisher","DOI":"10.1002\/j.1460-2075.1994.tb06651.x"},{"key":"e_1_2_1_46_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.260460502"},{"key":"e_1_2_1_47_1","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/5.2.175"},{"key":"e_1_2_1_48_1","first-page":"20216","article-title":"Biosynthesis of N\u2010glycolyneuraminic acid \u2014 the primary site of hydroxylation of N\u2010acetylneuraminic acid is the cytosolic sugar nucleotide pool","volume":"264","author":"Muchmore EA","year":"1989","journal-title":"J Biol Chem"},{"key":"e_1_2_1_49_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.95.16.9140"},{"key":"e_1_2_1_50_1","doi-asserted-by":"publisher","DOI":"10.1093\/oxfordjournals.jbchem.a124721"},{"key":"e_1_2_1_51_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0290(19990205)62:3<336::AID-BIT10>3.0.CO;2-N"},{"key":"e_1_2_1_52_1","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/8.5.445"},{"key":"e_1_2_1_53_1","doi-asserted-by":"publisher","DOI":"10.1042\/bj3050865"},{"key":"e_1_2_1_54_1","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/10.5.477"},{"key":"e_1_2_1_55_1","doi-asserted-by":"publisher","DOI":"10.1038\/40677"},{"key":"e_1_2_1_56_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.260440207"},{"key":"e_1_2_1_57_1","doi-asserted-by":"publisher","DOI":"10.1006\/abio.1997.2036"},{"key":"e_1_2_1_58_1","doi-asserted-by":"publisher","DOI":"10.1097\/00007890-199707270-00003"},{"key":"e_1_2_1_59_1","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/8.8.841"},{"key":"e_1_2_1_60_1","doi-asserted-by":"crossref","first-page":"6225","DOI":"10.1016\/S0021-9258(19)39314-7","article-title":"Transfer and expression of a murine UDP\u2010gal\u2010beta\u2010D\u2010gal\u2010alpha\u20101,3\u2010galactosyltransferase gene in transfected Chinese hamster ovary cells \u2014 competition reactions between the alpha\u20101,3\u2010galactosyltransferase and the endogenous alpha\u20102,3\u2010sialyltransferase","volume":"265","author":"Smith DF","year":"1990","journal-title":"J Biol Chem"},{"key":"e_1_2_1_61_1","doi-asserted-by":"publisher","DOI":"10.1006\/abio.1994.1138"},{"key":"e_1_2_1_62_1","doi-asserted-by":"publisher","DOI":"10.1038\/321618a0"},{"key":"e_1_2_1_63_1","doi-asserted-by":"publisher","DOI":"10.1038\/6179"},{"key":"e_1_2_1_64_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0290(19990820)64:4<401::AID-BIT3>3.0.CO;2-M"},{"key":"e_1_2_1_65_1","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/3.2.97"},{"key":"e_1_2_1_66_1","doi-asserted-by":"publisher","DOI":"10.1038\/15104"},{"key":"e_1_2_1_67_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi9621472"},{"key":"e_1_2_1_68_1","doi-asserted-by":"publisher","DOI":"10.1021\/bp9800945"}],"container-title":["Biotechnology and Bioengineering"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fbit.1051","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/bit.1051","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T20:11:04Z","timestamp":1760040664000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/bit.1051"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2001,3,2]]},"references-count":67,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2001,5,5]]}},"alternative-id":["10.1002\/bit.1051"],"URL":"https:\/\/doi.org\/10.1002\/bit.1051","archive":["Portico"],"relation":{},"ISSN":["0006-3592","1097-0290"],"issn-type":[{"value":"0006-3592","type":"print"},{"value":"1097-0290","type":"electronic"}],"subject":[],"published":{"date-parts":[[2001,3,2]]}}}