Invention Grant
US07642078B2 Methods to obtain recombinant proteins with increased sialylation from cells that express adenovirus E1A protein, and proteins obtained thereby
有权
从表达腺病毒E1A蛋白的细胞中获得具有增加的唾液酸化的重组蛋白的方法,以及由此获得的蛋白质
- Patent Title: Methods to obtain recombinant proteins with increased sialylation from cells that express adenovirus E1A protein, and proteins obtained thereby
- Patent Title (中): 从表达腺病毒E1A蛋白的细胞中获得具有增加的唾液酸化的重组蛋白的方法,以及由此获得的蛋白质
-
Application No.: US11793998Application Date: 2005-12-28
-
Publication No.: US07642078B2Publication Date: 2010-01-05
- Inventor: Dirk J. E. Opstelten
- Applicant: Dirk J. E. Opstelten
- Applicant Address: NL Leiden
- Assignee: Crucell Holland B.V.
- Current Assignee: Crucell Holland B.V.
- Current Assignee Address: NL Leiden
- Agency: TraskBritt
- International Application: PCT/EP2005/057205 WO 20051228
- International Announcement: WO2006/070011 WO 20060706
- Main IPC: C12N9/04
- IPC: C12N9/04 ; C12P21/06

Abstract:
The invention provides compositions comprising one or more isoforms of an erythropoietin (EPO) comprising glycans linked thereto, characterized in that said glycans comprise LewisX structures and on average at least 6 sialic acid moieties per EPO molecule. The invention further provides methods for obtaining a composition comprising one or more isoforms of an erythropoietin (EPO) comprising glycans linked thereto wherein said glycans comprise on average at least 6 sialic acids per EPO molecule and from 0 to 2 Lewis x structures, said method comprising: a) providing a eukaryotic cell containing a nucleic acid sequence encoding an adenoviral E1A protein in expressible format and further containing a nucleic acid encoding an EPO in expressible format, wherein said cell further contains a nucleic acid sequence encoding a sialyltransferase, preferably an alpha-2,6-sialyltransferase or an alpha-2,3-sialyltransferase, under control of a heterologous promoter; b) culturing said cell in a serum-free culture medium and allow expression of an EPO in said cell; c) harvesting the expressed EPO from said cell and/or from the culture medium; and d) purifying and fractionating the EPO to obtain fractions which have an increased average sialic acid content of the N-linked glycans per EPO molecule, to obtain a composition comprising one or more iso forms of an EPO comprising glycans linked thereto wherein said glycans comprise on average at least 6 sialic acids per EPO molecule and from 0 to 2 Lewis x structures.
Public/Granted literature
Information query