Abstract: Herein is reported a method for generating a recombinant mammalian cell expressing a heterologous polypeptide and a method for producing a heterologous polypeptide using said recombinant mammalian cell, wherein in the recombinant cell the expression of the endogenous SIRT-1 gene has been reduced. It has been found that the knockout of the sirtuin-1 gene (SIRT-1) in mammalian cells, e.g. such as CHO cells, improves recombinant productivity and reduces lactate production by the cells. Additionally, it has been found that the viability decline at the end of a fed-batch fermentation is reduced.

Abstract: Herein is reported a method for producing a bivalent, bispecific antibody comprising the steps of cultivating a mammalian cell comprising a deoxyribonucleic acid encoding the bivalent, bispecific antibody, and recovering the bivalent, bispecific antibody from the cell or the cultivation medium, wherein the deoxyribonucleic acid encoding the bivalent, bispecific antibody is stably integrated into the genome of the mammalian cell and comprises in 5?- to 3?-direction either a first expression cassette encoding the first light chain, a second expression cassette encoding the first heavy chain, a third expression cassette encoding the second light chain, and a fourth expression cassette encoding the second heavy chain, or a first expression cassette encoding the first light chain, a second expression cassette encoding the second heavy chain, a third expression cassette encoding the second light chain, and a fourth expression cassette encoding the first heavy chain, wherein the first heavy chain comprises from N- t

Abstract: Herein is reported a method for producing a recombinant mammalian cell comprising a deoxyribonucleic acid encoding a polypeptide and secreting the polypeptide comprising the steps of a) providing a mammalian cell comprising an exogenous nucleotide sequence integrated at a single site within a locus of the genome of the mammalian cell, wherein the exogenous nucleotide sequence comprises a first and a second recombination recognition sequence flanking at least one first selection marker, and a third recombination recognition sequence located between the first and the second recombination recognition sequence, and all the recombination recognition sequences are different; b) introducing into the cell provided in a) a composition of two deoxyribonucleic acids comprising three different recombination recognition sequences and one to eight expression cassettes, wherein the first deoxyribonucleic acid comprises in 5?- to 3?-direction, a first recombination recognition sequence, one or more expression cassette(s), a

Abstract: Herein is reported a method for generating a recombinant mammalian cell expressing a heterologous polypeptide and a method for producing a heterologous polypeptide using said recombinant mammalian cell, wherein in the recombinant cell the expression of at least the endogenous gene MYC has been reduced. It has been found that the knockout of at least the endogenous gene MYC in mammalian cells, e.g. such as CHO cells, improves recombinant productivity by the cells.

Abstract: Herein are reported novel DNA constructs and methods using the same. The current invention uses a deliberate arrangement of non-productive/inactive promoters and genes on coding and template strands of DNA molecules, which are converted into their active form by the interaction with a site-specific recombinase. In more detail, the DNA element according to the current invention is non-functional with respect to the expression of the contained first and second genes. By being non-functional with respect to the expression of the first and second gene, the DNA element according to the invention can be integrated into genome of a cell without the risk that the comprised structural genes are expressed already directly after the integration. The genes are only expressed once a recombinase recognizing and functional with the recombination recognition sequences of the DNA element is activated or introduced into the cell.

Abstract: Herein is reported a novel adenoviral VA RNA nucleic acid wherein the wild-type type 2 polymerase III promoter has been removed and an U6-snRNA promoter or an inducible promoter has been added.

Abstract: Disclosed herein, in part, are methods for the expression and production of a trivalent, bispecific antibody using a recombinant nucleic acid comprising multiple, different expression cassettes in a specific and defined sequence, which are stably integrated into the genome of a mammalian cell.by targeted integration.

Abstract: The invention relates to vectors and mammalian cells in a system useful for switching on or switching off gene expression in response to pH and gaseous carbon dioxide, taking advantage of signal transduction of the human proton-activated cell-surface receptor TDAG8, OGR1 and/or GPR4 and related receptors to chimeric promoters containing cAMP-response elements. The systems according to the invention can be used to precisely monitor culture pH within the physiologic range, or also to adjust expression by gaseous C02 which shifts the C02-bicarbonate balance towards hydrogen ions. Production of biopharmaceuticals of engineered cells cultivated in a bioreactor setup can be induced by gaseous C02. Also, implanting engineered cells provide novel treatment strategies for acidosis-related disorders and type 1 diabetes.