Taci-immunoglobulin fusion proteins

A technology of immunoglobulin and fusion protein, applied in the field of improving TACI-immunoglobulin fusion protein

Inactive Publication Date: 2010-01-20
ZYMOGENETICS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The original version resulted in low-level expression of heterogeneous proteins

Method used

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  • Taci-immunoglobulin fusion proteins
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  • Taci-immunoglobulin fusion proteins

Examples

Experimental program
Comparison scheme
Effect test

example TA

[0102] Example TACI-Fc fusion protein constructs

[0103]

TACI sequence a

Fc version

TACI b

Fc4

TACI b

Fc5

TACI b

Fcγ1

TACI (d107-154)

Fc5

TACI (R119Q)

Fc4

TACI(1-104)-BCMA(42-54) c

Fc5

TACI (d143-150)

Fc5

TACI (R142G, d143-150)

Fc5

TACI (R119G, Q121P, R122Q, S123A)

Fc5

TACI (R119G, R122Q)

Fc5

TACI (d1-28, V29M)

Fc6

TACI (d1-29)

Fc6

TACI (d1-29)

Fc5

TACI (d1-29, d107-154)

Fc5

TACI (d1-29, d111-154)

Fc5

TACI (d1-29, d120-154)

Fc5

[0104] a Provides position, mutation and deletion information on the amino acid sequence with reference to the amino acid sequence of SEQ ID NO: 2 in parentheses.

[0105] b includes amino acid residues 1 to 154 of SEQ ID NO:2.

[0106] c This construct includes amino acid residues 1 to 104 of SEQ ID NO: 2 (TACI) and amino acids 42 to 5...

Embodiment 1

[0140] Example 1 describes an expression vector comprising a cytomegalovirus promoter directing transgenic expression of a recombinant protein, an immunoglobulin intron, and a tissue plasminogen activator signal sequence. A suitable immunoglobulin intron is the murine 26-10V H Intron. SEQ ID NO: 66 provides mouse 26-10V H Exemplary nucleotide sequences of introns. The expression vector may also include a 5' untranslated region (UTR) upstream of the nucleotide sequence encoding the TACI-immunoglobulin protein. Suitable 5'-UTR can be from mouse 26-10V H obtained in the gene. SEQ ID NO: 63 discloses useful natural mouse 26-10V H The 5'-UTR nucleotide sequence, while SEQ ID NO: 64 shows the mouse 26-10V H When using the 5'-UTR nucleotide sequence, it has been optimized at the 3' end.

[0141] As an example, SEQ ID NO: 67 provides the nucleotide sequence comprising the following elements: native murine 26-10V H 5'-UTR (nucleotides 1 to 51), mouse 26-10V H Signal sequence...

Embodiment 2

[0301] Production of TACI-Fc Protein by Chinese Hamster Ovary Cells

[0302] By means of electroporation, the TACI-Fc expression construct was used to transfect suspension-adapted Chinese hamster ovary (CHO) DG44 cells grown in animal protein-free medium (Urlaub et al., Som. Cell. Molec. Genet. 12:555 (1986)). CHO DG44 cells lack a functional dihydrofolate reductase gene due to deletions at two dihydrofolate reductase chromosomal locations. Growth of the cells in the presence of increasing concentrations of methotrexate results in amplification of the dihydrofolate reductase gene and the linked gene encoding the recombinant protein on the expression construct.

[0303] CHO DG44 cells were passaged in PFCHO medium (JRH Biosciences, Lenexa, KS), 4mM L-glutamine (JRH Biosciences), and 1x hypothanxine-thymidine rehydration solution (Life Technologies), and then incubated at 37°C and 5% CO 2 The cells were incubated in Corning shaker flasks on a rotary shaker platform rotating at...

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Abstract

Molecules that interfere with the binding of a tumor necrosis factor receptor with its ligand, such as a soluble receptor, have proven usefulness in both basic research and as therapeutics. The present invention provides improved soluble transmembrane activator and calcium modulator and cyclophilin ligand-interactor (TACI) receptors.

Description

[0001] This application is a divisional application of an invention patent application with an application date of May 20, 2002, an application number of 02812698.X, and an invention title of "TACI-immunoglobulin fusion protein". technical field [0002] The present invention generally relates to improved fusion proteins comprising a tumor necrosis factor receptor portion and an immunoglobulin portion. In particular, the invention relates to improved TACI-immunoglobulin fusion proteins. Background of the invention [0003] Cytokines are small soluble proteins that mediate growth and various biological effects in many cell types, (see, eg, Arai et al., Annu. Rev. Biochem. 59:783 (1990); Mosmann, Curr. Opin . Immunol. 3:311 (1991); Paul and Seder, Cell 76:241 (1994)). The proteins that make up the cytokine group include interleukins, interferons, colony-stimulating factors, tumor necrosis factors, and other regulatory molecules. For example: human interleukin-17 is a cytokin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/17A61K39/395A61P35/00C07K19/00C12N15/62C12N15/09A61K38/00A61K48/00C07K14/705C07K14/715C07K16/00
CPCC07K16/241C07K2319/00C07K14/70578A61K38/00C07K2319/30A61P1/00A61P11/00A61P11/06A61P13/12A61P17/06A61P19/00A61P19/02A61P21/04A61P25/00A61P29/00A61P31/04A61P35/00A61P35/02A61P37/00A61P37/02A61P37/06A61P7/00A61P7/06A61P7/10A61P9/00A61P3/10A61K39/395A61K39/3955A61K45/06
Inventor M·W·里克森J·A·格罗斯
Owner ZYMOGENETICS INC
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