Compositions and Methods of Use for Mgd-Csf in Disease Treatment

Inactive Publication Date: 2007-11-15
FIVE PRIME THERAPEUTICS
View PDF1 Cites 12 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0043] MGD-CSF promotes the proliferation, survival, and/or differentiation of monocytes, granulocytes, dendritic cells, and NK cells. Therefore, MGD-CSF finds use as a protein therapeutic for treating cancers through its ability to stimulate the proliferation and/or activation of immune cells such as NK cells, monocytes, macrophages, granulocytes, and dendritic cells to fight tumor cells. MGD-CSF may be used alone or in combination with therapeutic monoclonal antibodies (for example, Rituxan) to treat cancer, since MGD-CSF may promote antibod

Problems solved by technology

Similarly, humans with naturally occurring NK cell deficien

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Compositions and Methods of Use for Mgd-Csf in Disease Treatment
  • Compositions and Methods of Use for Mgd-Csf in Disease Treatment
  • Compositions and Methods of Use for Mgd-Csf in Disease Treatment

Examples

Experimental program
Comparison scheme
Effect test

example 1

Amino Acid Sequence Alignment of MGD-CSF with MCG34647

[0378] MGD-CSF and NP—689669 were compared by aligning their amino acids using the program clustal format for T-COFFEE Version 1.37, cpu=0.00 sec., score=72, Nseq=3, len=242. As shown in FIG. 1, the amino acid sequence of MGD-CSF differs from the amino acid sequence of NP—689669 (MCG34647). The latter sequence has a glutamine (Q) residue at amino acid 81. The five flanking amino acid residues adjacent to and on either side of amino acid 81 in the NCBI sequence of MCG34647 are NVTRLQRAQVS (SEQ ID NO.:279). In contrast to this published sequence of MCG34647, MGD-CSF contains the amino acid sequence NVTRLRAQVS (SEQ ID NO.:280). The difference between these sequences results from alternative splicing of the MCG34647 gene between exons 3 and 4. The genome sequences at the exon 3-4 boundary are the codons aac gtc acc agg ctg gtg (SEQ ID NO.:281) and cag cag agg gcc cag gtg agc (SEQ ID NO.:282), wherein the gtg codon (shown in italics)...

example 2

Plasmid Vectors for MGD-CSF Expression

[0380] The MGD-CSF gene was cloned into pTT-5 and pTT-2 mammalian expression vectors modified as shown in FIG. 2 and FIG. 3 using standard cloning procedures. The MGD-CSF gene was also cloned into the pIB / V5His-DEST insect cell expression vector (Invitrogen, Carlsbad Calif.) using standard cloning procedures. The resulting constructs are described in Table 1 and Table 5. They include human MGD-CSF untagged in vector pTT5 (MGD-CSF), human MGD-CSF untagged in vector pTT2 (CLN00839395), human MGD-CSF with a C-terminal V5H8 tag in vector pTT5 (CLN00732663), human MGD-CSF tagged with V5H8 (CLN00732663), human MGD-CSF with a C-terminal V5H8 tag in vector pTT2 (CLN00840351), human MGD-CSF with a C-terminal V5H8 tag in vector pIB / V5His-DEST (CLN00758593), and human MGD-CSF with a collagen secretory leader and a C-terminal Streptag in vector pTT5-G (CLN00816424).

[0381] To monitor the expression and secretion of MGD-CSF and to aid in its purification, t...

example 3

Transient Expression in Mammalian Cells

[0387] Complementary DNA encoding the MGD-CSF polypeptide was cloned into the expression vectors pTT5 and pCDNA-pDEST40 and expressed as both a tagged and untagged protein. Protein levels were quantified by measuring the levels of the tag, for example, a V5His tag, by quantitative Western blot analysis. The expression vectors were transfected into adherent 293T cells using the transfection agent Fugene 6 (Roche, Nutley N.J.) in DMEM with 10% fetal bovine serum (FBS) and penicillin / streptomycin (100 μg / ml, 100 U / ml), and incubated at 37° C. in 5% CO2 for 40 hours, after which the cells were washed with PBS and incubated for an additional 48 hours in complete DMEM. Cell supernatant was harvested, cleared of cell debris by centrifugation, and tested for biological activity (untagged cDNA) and protein expression (V5 tagged cDNA) by Western blot assay using an anti-V5 antibody.

[0388] Expression studies were also performed with 293-6E cells transie...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Volumeaaaaaaaaaa
Volumeaaaaaaaaaa
Volumeaaaaaaaaaa
Login to view more

Abstract

Disclosed is a newly identified secreted molecule, identified herein as “monocyte, granulocyte, and dendritic cell colony stimulating factor” (MGD-CSF), the polypeptide sequence, and polynucleotides encoding the polypeptide sequence. Also provided is a procedure for producing the polypeptide by recombinant techniques employing, for example, vectors and host cells. Additionally, procedures are described to modify the disclosed novel molecules of the invention to prepare fusion molecules. Also disclosed are methods for using the polypeptides and active fragments thereof for treatment of a variety of diseases, including, for example, cancer, autoimmune and inflammatory diseases, infectious diseases, and recurrent pregnancy loss.

Description

PRIORITY CLAIM [0001] This application claims priority to provisional applications 60 / 590,565, filed Jul. 22, 2004; 60 / 647,604, filed Jan. 27, 2005; 60 / 664,932, filed Mar. 24, 2005; and a provisional application entitled “Novel MGD-CSF Polypeptides, Polynucleotides, and Methods of Use Thereof,” filed Jul. 14, 2005. This application also relates to applications PCT / US03 / 34811, filed Oct. 31, 2003; PCT / US04 / 11270, filed Apr. 30, 2004; 60 / 590,565 filed Jul. 22, 2004; 60 / 642,604, filed Jan. 11, 2005; 60 / 647,013, filed Jan. 27, 2005; and 60 / 654,229, filed Feb. 18, 2005. These applications are all incorporated by reference in their entireties.TECHNICAL FIELD [0002] The present invention relates to a novel secreted molecule identified herein as “monocyte, granulocyte, and dendritic cell colony stimulating factor” (MGD-CSF). It relates to the polypeptide and polynucleotide sequences of MGD-CSF, fusion molecules containing MGD-CSF, vectors, host cells, compositions, and kits comprising MGD-C...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): A61K39/00A61K38/00C07H21/00C07K14/00C07K16/18C12N15/00C12N15/87C12N5/06C12P1/00G01N33/53
CPCA61K38/00C07K16/243C07K14/53A61P1/04A61P1/14A61P1/16A61P15/00A61P15/06A61P17/06A61P19/02A61P25/00A61P29/00A61P31/00A61P31/04A61P31/10A61P31/12A61P33/00A61P35/00A61P35/02A61P37/00A61P37/02A61P37/04A61P37/06A61P9/00A61P9/10
Inventor BEHRENS, DIRKBOSCH, ELIZABETHDOBERSTEIN, STEPHEN K.HALENBECK, ROBERT FORGANHESTIR, KEVINHUANG, MIN MEILEE, ERNESTINELIN, HAISHANLINNEMANN, THOMASMARSHALL, SHANNONWONG, JUSTIN G. P.WU, GEZHOU, AILEEN
Owner FIVE PRIME THERAPEUTICS
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap