Hoffmann-la Roche Inc.

Roche Palo Alto LLC

F. Hoffmann-la Roche Ag

Roche Glycart Ag

Hendrik Knoetgen

Christian Klein

Jens Niewoehner

Monika Baehner

Erhard Kopetzki

Joerg Thomas Regula

Kurt Lang

Ulrich Goepfert

Christian Schantz

Andreas Schaubmar

Anne Stern

Ekkehard Moessner

Pablo Umana

Stephan Fischer

Friederike Hesse

Guy Georges

Peter Michael Huelsmann

Friedrich Metzger

Hans-Joachim Schoenfeld

Sabine Imhof-Jung

Hans-Willi Krell

Nikolaos Dimoudis

Johannes Auer

Silke Hansen

Oliver Ploettner

Dirk Ponsel

Sandra Miller

Engin Toksoez

Martina Schmittnaegel

Petra Hanke

Claire Louise Langrish

Hans Joachim Schoenfeld

Herein is reported an expression vector comprising &#x2014;an antibody light chain expression cassette, &#x2014;an antibody heavy chain expression cassette, and &#x2014;a selection marker expression cassette, wherein the expression cassettes are arranged unidirectional, and wherein the expression cassettes are arranged in the 5' to 3&#x2032; sequence of antibody heavy chain expression cassette, antibody light chain expression cassette and selection marker expression cassette. Further are reported herein methods for the generation of antibody producing cells and the use of these cells for the recombinant production of antibodies.

Expression vector organization, novel production cell generation methods and their use for the recombinant production of polypeptides

Herein is reported a disulfide-linked multivalent multi-function protein, characterized in that it comprises two or more antigen presenting domains, exactly one antibody Fc-region, and at least one antigen binding site, wherein the antigen presenting domain comprises in N- to C-terminal direction either (i) a &#x3b2;2-microglobulin, and (ii) the extracellular domains &#x3b1;1, &#x3b1;2, and &#x3b1;3 of a class I MHC molecule with a relative frequency of less than 1%, or (i) a T-cell response eliciting peptide, (ii) a &#x3b2;2-microglobulin, and (iii) the extracellular domains &#x3b1;1, &#x3b1;2, and &#x3b1;3 of a class I MHC molecule with a relative frequency of 1% or more, wherein the antigen binding site binds to a cancer cell surface antigen or a virus-infected cell surface antigen and wherein the antigen presenting domain has at least two non-naturally occurring cysteine residues which form an intrachain/interdomain disulfide bond.

Disulfide-linked multivalent MHC class I comprising multi-function proteins

Herein is reported an expression vector comprising&#x2014;an antibody light chain expression cassette, &#x2014;an antibody heavy chain expression cassette, and&#x2014;a selection marker expression cassette, wherein the expression cassettes are arranged unidirectional, and wherein the expression cassettes are arranged in the 5' to 3&#x2032; sequence of antibody heavy chain expression cassette, antibody light chain expression cassette and selection marker expression cassette. Further are reported herein methods for the generation of antibody producing cells and the use of these cells for the recombinant production of antibodies.

The current invention reports a method for the recombinant production of a secreted heterologous immunoglobulin in a CHO cell comprising the following steps: i) providing a CHO cell, which is adapted to growth in suspension culture, adapted to growth in serum-free medium, mycoplasma free, and virus free, ii) providing a vector comprising a prokaryotic origin of replication, a first nucleic acid conferring resistance to a prokaryotic selection agent, a second nucleic acid encoding the heavy chain of said heterologous immunoglobulin, a third nucleic acid encoding the light chain of said heterologous immunoglobulin, a fourth nucleic acid conferring resistance to a eukaryotic selection agent, iii) transfecting said CHO cell, wherein said transfecting comprises a) transfecting said CHO cell with said vector comprising a fourth nucleic acid conferring resistance to a first eukaryotic selection agent, b) selecting a CHO cell by growth in cultivation medium containing said first eukaryotic selection agent, c) transfecting said selected CHO cell with said vector comprising a fourth nucleic acid conferring resistance to a second eukaryotic selection agent different to said first eukaryotic selection agent, d) selecting a CHO cell by selected growth in cultivation medium containing said first and said second eukaryotic selection agent, iv) cultivating said transfected CHO cell in a medium in the presence of said first and second eukaryotic selection agent, under conditions suitable for the expression of said second, and third nucleic acid, and v) recovering said secreted heterologous immunoglobulin from the cultivation medium.

Protein expression from multiple nucleic acids

The invention provides antibodies binding to TWEAK, including anti-TWEAK antibodies comprising a heavy chain variable domain CDR3 (CDR3H) selected from the group consisting of SEQ ID NO: 8, 16 or 24. The invention provides anti-TWEAK antibodies which are useful for the treatment of cancer, autoimmune diseases, rheumatoid arthritis, psoratic arthritis, muscle diseases, e.g. muscular dystrophy, multiple sclerosis, chronic kidney diseases, bone diseases, e.g. bone degeneration in multiple myeloma, systemic lupus erythematosus, lupus nephritis, and vascular injury.

Antibodies against human TWEAK and uses thereof

An antibody binding to TWEAK comprising as heavy chain variable domain a CDR3H selected from the group consisting of SEQ ID NO: 8, 16 or 24.

Antibodies against human tweak and uses thereof

Method for the production of IGF-I, characterized by cultivating a prokaryotic host cell comprising an expression vector containing a nucleic acid encoding a fusion protein comprising said IGF-I N-terminally linked to the C-terminus of a propeptide, whereby said propeptide ends C-terminally with amino acids -Y-Pro, wherein Y is selected from the group consisting of Pro, Pro-Ala, Pro-Gly, Pro-Thr, Ala-Pro, Gly-Pro, Thr-Pro, Arg-Pro, or Pro-Arg-Pro, recovering and cleaving said fusion protein with IgA protease, and recovering said IGF-I. IGF-I is useful for the treatment of neurodegenerative disorders like Alzheimer's Disease.

Method for the production of insulin-like growth factor-1

The present invention relates to a fusion protein comprising IGF-I or an IGF-I variant N-terminally linked to the C-terminus of a propeptide. The invention relates also to a method involving the use of the aforementioned fusion protein in the production of a lysine-PEGylated IGF-I or IGF-I variant. The method comprises the steps of cultivating a prokaryotic host cell comprising an expression vector containing a nucleic acid encoding the fusion protein and causing the cell to express the fusion protein, recovering and PEGylating said fusion protein, cleaving said PEGylated fusion protein with IgA protease, and recovering lysine-PEGylated IGF-I or IGF-I variant. The invention relates also to a lysine-PEGylated IGF-I or IGF-I variant produced using the above method. In addition, the invention relates to a method for treating a neurodegenerative disorders like Alzheimer's Disease using the lysine-PEGylated IGF-I or IGF-I variant and a composition comprising the lysine-PEGylated IGF-I or IGF-I variant.

Method for the production of conjugates of insulin-like growth factor-1 and poly(ethylene glycol)

The current invention comprises a nucleic acid encoding the amino acid sequence of the C-terminal part of the C3-domain of an immunoglobulin of the class IgA or IgG, or the C-terminal part of the C4-domain of an immunoglobulin of the class IgE or IgM, wherein the glycine-lysine-dipeptide comprised in the amino acid sequence of the C-terminal part of the C3- or C4-domain is encoded by the nucleic acid ggaaaa, or the nucleic acid ggcaaa, or the nucleic acid gggaaa, or the nucleic acid gggaag, or the nucleic acid ggcaag, or the nucleic acid ggaaag.

Heavy chain mutant leading to improved immunoglobulin production

Disclosed herein are methods for the recombinant expression of mammalian cell membrane-bound human CCN1 or a CCN1 domain thereof in mammalian cells, comprising transforming a mammalian host cell with a vector encoding CCN1 or a CCN1 domain thereof C-terminally fused to a mammalian cell transmembrane domain (CCN1 fusion protein), expressing said CCN1 or CCN1 domain fusion protein in said host cell, and recovering said membrane bound CCN1 or said CCN1 domain thereof. Disclosed herein is the use of such membrane bound CCN1 and domain for the generation of corresponding antibodies. Antibodies against human CCN1 are useful for the treatment of diseases.

Antibodies against human CCN1 and uses thereof

An antibody binding to IL-17 characterized by binding to the same IL-17 epitope to which monoclonal antibody 3C1 binds, and being of human IgG1 isotype modified in the hinge region at amino acid position 216-240, preferably at amino acid position 220-240, between C1 and C2 and/or in the second inter-domain region at amino acid position 327-331 between C2 and C3 has advantageous properties for the treatment of inflammatory diseases.

Antibodies against human IL17 and uses thereof

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Penzberg, Germany

Hendrik Knoetgen