Title:
METHOD FOR PREPARING RECOMBINANT GLYCOPROTEINS WITH HIGH SIALIC ACID CONTENT
Document Type and Number:
WIPO Patent Application WO/2011/096750
Kind Code:
A3
Abstract:
The present disclosure relates to a method for preparing recombinant glycoproteins with high sialic acid content. More specifically, for UDP-GlcNAc 2-epimerase/ManNAc kinase (GNE/MNK) enzyme where point mutation was induced by substituting arginine at position 263 by leucine only or by further substituting arginine at position 266 by glutamine, epimerase activity is constantly maintained, and overexpressed cells thereof experience an increase in intracellular cytidine monophosphate (CMP)-sialic acid content, irrespective of CMP-sialic acid concentration. Particularly, since in an glycoprotein(such as, erythropoietin and thrombopoietin)-producing host cell where point mutationinduced GNE/MNK, human alpha-2,3-sialyltransferase and a CMP-sialic acid transporter gene are simultaneously overexpressed, intracellular content of CMP-sialic acid and sialic acid in glycoprotein increases in cells, overexpression in a host cell producing a sialylated recombinant glycoprotein the three genes above may be useful for preparing glycoprotein with increased sialic acid content.
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Inventors:
KIM JUNG HOE (KR)
SON YOUNG DOK (KR)
HWANG JIN YOUNG (KR)
JEONG YEON TAE (KR)
SON YOUNG DOK (KR)
HWANG JIN YOUNG (KR)
JEONG YEON TAE (KR)
Application Number:
PCT/KR2011/000755
Publication Date:
January 05, 2012
Filing Date:
February 01, 2011
Export Citation:
Assignee:
KOREA ADVANCED INST SCI & TECH (KR)
KIM JUNG HOE (KR)
SON YOUNG DOK (KR)
HWANG JIN YOUNG (KR)
JEONG YEON TAE (KR)
KIM JUNG HOE (KR)
SON YOUNG DOK (KR)
HWANG JIN YOUNG (KR)
JEONG YEON TAE (KR)
International Classes:
C12N15/54; C12N15/18; C12N15/79; C12P21/02
Foreign References:
| US20020045207A1 | 2002-04-18 | |||
| US20090298120A1 | 2009-12-03 | |||
| US6949372B2 | 2005-09-27 |
Other References:
JAY ET AL.: "Preclinical assessment of wt GNE gene plasmid for management of hereditary inclusion body myopathy 2 (HIBM2)", GENE REGUL SYST BIO., vol. 2, 20 June 2008 (2008-06-20), pages 243 - 252, XP002650019
SEPPALA ET AL.: "Mutations in the human UDP-N-acetylglucosamine 2-epimerase gene define the disease sialuria and the allosteric site of the enzyme", AM J HUM GENET., vol. 64, no. 6, June 1999 (1999-06-01), pages 1563 - 1569, XP002171406
GALEANO ET AL.: "Mutation in the key enzyme of sialic acid biosynthesis causes severe glomerular proteinuria and is rescued by N-acetylmannosamine", J CLIN INVEST, vol. 117, no. 6, June 2007 (2007-06-01), pages 1585 - 1594, XP008151028
See also references of EP 2534249A4
SEPPALA ET AL.: "Mutations in the human UDP-N-acetylglucosamine 2-epimerase gene define the disease sialuria and the allosteric site of the enzyme", AM J HUM GENET., vol. 64, no. 6, June 1999 (1999-06-01), pages 1563 - 1569, XP002171406
GALEANO ET AL.: "Mutation in the key enzyme of sialic acid biosynthesis causes severe glomerular proteinuria and is rescued by N-acetylmannosamine", J CLIN INVEST, vol. 117, no. 6, June 2007 (2007-06-01), pages 1585 - 1594, XP008151028
See also references of EP 2534249A4
Attorney, Agent or Firm:
LEE, Won-Hee (642-16 Yoksam-dong Kangnam-ku, Seoul 135-080, KR)
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