Multimerization of recombinant protein by fusion to a sequence from lamprey 机翻标题: 暂无翻译,请尝试点击翻译按钮。

公开号/公开日
IN201747025417 A 2017-08-11 [IN201747025417] / 2017-08-11
申请号/申请日
2017IN-47025417 / 2017-07-18
发明人
SODOYER Régis;LEGASTELOIS Isabelle;
申请人
SANOFI PASTEUR;
主分类号
IPC分类号
C07K-019/00C12N-015/62
摘要
(IN201747025417) The present invention relates to polymerized recombinant proteins to recombinant nucleic acids coding for the polymerized recombinant proteins to expression cassettes comprising the recombinant nucleic acids to host cells transformed by the expression cassettes and to a method for multimerizing a recombinant protein.  The polymerized proteins of the invention may be used in pharmaceutical or immunogenic compositions.  In particular the recombinant proteins may be antigens antibodies or scaffolds.  In particular the polymerized recombinant protein may be an influenza haemagglutinin.
机翻摘要
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地址
代理人
代理机构
;
优先权号
2014EP-0307096 2014-12-19
主权利要求
(IN201747025417)  1. A molecule which comprises a first amino acid sequence which has at least 80% identity to SEQ ID NO: 1 and a second amino acid sequence which is heterologous to said first sequence, wherein said molecule does not comprise a ieucine-rich repeat (LRR) module from a lamprey VLR-B antibody.  10. 2. A molecule according to claim 1, wherein said molecule does not comprise a sequence selected from the group of sequences defined by SEQ ID NO: 29.  15. 3. A molecule according to claim 1 or 2, wherein the only amino acid sequence in said molecule which is derived from a lamprey VLR-B antibody is the sequence having at least 80% identity to SEQ ID NO: 1. 4. The molecule according to any one of claims 1 to 3 wherein said 20. Molecule is a recombinant protein. 5. The molecule according to any one of claims 1 to 4 which comprises cysteine residues at the positions within the molecule corresponding to positions 2, 7, 13, 19,21, 24 and 27 of SEQ IDNO:1. 6. The molecule according to any one of claims 1 to 5 wherein the first amino acid sequence has at least 90% identity or 100% identity to SEQ !D NO: 1. 7. The molecule according to any one of c/aims 1 to 6 which comprises SEQ ID NO: 2.  5. 8. The molecule according to any one of claims 1 to 7, wherein there is a linker between the first amino acid sequence and the heterologous amino acid sequence.  10. 9. The molecule according to any one of claims 1 to 8, wherein the heterologous amino acid sequence encodes an antigen. 10. The molecule of claim 9 wherein the antigen is selected from the group consisting of influenza virus, HIV, cytomegalovirus, dengue virus, yellow 15. Fever virus, tick-borne encephalitis virus, hepatitis virus, Japanese encephalitis virus, human papillomavirus, coxsackievirus, herpes simplex virus, rubella virus, mumps virus, measles virus, rabies virus, polio virus, rotavirus, respiratory syncytial virus, Ebola virus, Chikungunya virus, Mycobacterium tuberculosis, Staphylococcus aureus, Staphylococcus epidermidis, E. coli, Clostridium difficile, 20. Bordeteila pertussis, Clostridium tetani, Haemophilus influenzae type b, Chlamydia pneumoniae, Chlamydia trachomatis, Porphyromonas gingivalis, Pseudomonas aeruginosa, Mycobacterium diphtheriae, Shigella, Neisseria meningitidis, Streptococcus pneumoniae and Plasmodium falciparum. 11. The molecule of claim 10, wherein the antigen is from influenza virus and is selected from the group consisting of a haemaglutinin (HA), a matrix 2 protein (M2), and an HAM2 fusion protein.  5. 12. The molecule of claim 11, wherein the antigen is an influenza haemaglutinin, preferably the ectodomain of an influenza haemaglutinin. 13. The molecule of claim 10, wherein the antigen is from Shigella and Is selected from the group consisting of IpaD and MxiH.  10. 14. The molecule according to any one of claims 1 to 8, wherein the heterologous amino acid sequence encodes an antibody or a scaffold. 15. The molecule of claim 14 wherein the antibody or scaffold is 15. Specific for an antigen selected from the group consisting of influenza virus, HIV, Cytomegalovirus, dengue virus, yellow fever virus, tick-borne encephalitis virus, hepatitis virus, Japanese encephalitis virus, human papillomavirus, coxsackievirus, herpes simplex virus, rubella virus, mumps virus, measles virus, rabies virus, polio virus, rotavirus, respiratory syncytial virus, Ebola virus, 20. Chikungunya virus, Mycobacterium tuberculosis, Staphylococcus aureus, Staphylococcus epidermidis, E. coli, Clostridium difficile, Bordeteila pertussis, Clostridium tetani, Haemophilus influenzae type b, Chlamydia pneumoniae, Chlamydia trachomatis, Porphyromonas gingivaiis, Pseudomonas aeruginosa, Mycobacterium diphtheriae, Shigella, Neisseria meningitidis, Streptococcus 25. Pneumoniae and Plasmodium falciparum. 16. The molecule of claims 14 or 15 wherein the antibody is selected from the group consisting of a monoclonal antibody, a single domain antibody (dAb), a single-chain variable fragment (scFv), a Fab, a F(ab')2 and a diabody 5. (Db). 17. The molecule of claims 14 or 15 wherein the heterologous amino acid sequence encodes an antibody or scaffold selected from the group consisting of a bi-specific antibody, a multi-specific antibody, a bi-specific 10. Scaffold, and a multi-specific scaffold. 18. A recombinant nucleic acid which comprises a first nucleic acid sequence with at least 80% identity to SEQ ID NO;  3 and a second nucleic acid sequence which is heterologous to said first sequence, wherein said recombinant 15. Nucleic acid does not encode a leucine-rich repeat (LRR) module from a lamprey VLR-8 antibody. 19. The recombinant nucleic acid of claim 18 wherein said first nucleic acid sequence encodes an amino acid sequence which comprises cysteine 20 residues at positions within said amino acid sequence that correspond to positions 2, 7( 13, 19, 21, 24 and 27 of SEQ ID NO:1. 20. The recombinant nucleic acid of claims 18 or 19 wherein the first nucleic acid sequence has at least 90% identity or 100% identity to SEQ ID NO: 21. The recombinant nucleic acid of claims 18 or 19 which comprises SBQ )D HO: 4.  5. 22. An expression cassette comprising a recombinant nucleic acid as claimed in any one of claims 18 to 21 and wherein the recombinant nucleic acid is operably linked to a promoter, 23. A host cell transformed with an expression cassette as claimed in 10 claim 22. 24. The host cell of claim 23 wherein the host cell is a prokaryote, Preferably a prokaryote host cell selected from the group consisting of E.co/i, a Bacillus species, Lactococcus lactis, Pseudomonas fluorescens, a Caulobacter 15. Species, Corynebacterium glutamicum and Ralstonia eutropha. 25. The host cell of claim 23 wherein the host ceil is an eukaryote, Preferably a host ceff selected from the group consisting of a protist, an insect cell, a yeast, a mammalian cell, a plant cell, a micro-algae or a fungus..  20. 26. The host cell of claim 23 wherein the host cell is selected from the group consisting of Leishmania tarentolae, CHO and E. coli. 27. The molecule as claimed in any one of claims 1 to 17, which is 25. Capable of forming a stable multimeric protein. 28. A stable homo-multimeric recombinant protein which comprises a protein selected from the group consisting of the ectodomain of an influenza HA protein, a Shigella IpaD protein and a Shigella MxiH protein, fused to a protein having an amino acid sequence with at least 80% identity to SEQ ID NO: 1. 29. A pharmaceutical composition comprising a molecule as claimed in any one of cfaims 1 to 17 or 27, or a recombinant protein as claimed in claim 28, and a pharmaceuticaily acceptable carrier or diluent. 30. The molecule as claimed in anyone of claims 1 to 17 or 27, or the recombinant protein as claimed in claim 28, for use as a medicament. 31. The molecule as claimed in any one of claims 9 to 13, or the recombinant protein as claimed in claim 28, for use in inducing an immune response to an antigen in a subject. 32. A method for multimerizing a recombinant protein comprising: A) fusing a nucleic acid sequence having at least 80% identity to SEQ ID NO: 3 to the nucleic acid sequence coding for said recombinant protein, with the proviso that said recombinant protein does not comprise a leucine-rich repeat (LRR) module from a lamprey VLR-B antibody, B) expressing the fusion protein encoded by said nucleic acid sequence, under conditions which lead to the multimerization of said recombinant protein.
法律状态
(IN201747025417) LEGAL DETAILS FOR IN201747025417  Actual or expected expiration date=2037-07-18    Legal state=ALIVE    Status=PENDING     Event publication date=2017-07-18  Event code=IN/APP  Event indicator=Pos  Event type=Examination events  Application details  Application country=IN IN201747025417  Application date=2017-07-18  Standardized application number=2017IN-47025417     Event publication date=2017-08-11  Event code=IN/A  Event indicator=Pos  Event type=Examination events  Application laid open  Publication country=IN  Publication number=IN201747025417  Publication stage Code=A  Publication date=2017-08-11  Standardized publication number=IN201747025417
专利类型码
A
国别省市代码
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