Abstract: Herein is reported a method for producing a trivalent antibody comprising the steps of cultivating a mammalian cell comprising a deoxyribonucleic acid encoding the trivalent antibody, and recovering the trivalent antibody from the cell or the cultivation medium, wherein the deoxyribonucleic acid encoding the trivalent antibody is stably integrated into the genome of the mammalian cell and comprises in 5?- to 3?-direction a first expression cassette encoding the first heavy chain, a second expression cassette encoding the first light chain, a third expression cassette encoding the first light chain, a fourth expression cassette encoding the second heavy chain, a fifth expression cassette encoding the second light chain, and a sixth expression cassette encoding the second light chain, wherein the first heavy chain comprises from N- to C-terminus a first heavy chain variable domain, a CH1 domain, a first light chain variable domain, a CH1 domain, a hinge region, a CH2 domain and a CH3 domain, the second heavy ch

Abstract: Herein is reported a method for producing a multivalent, bispecific antibody comprising the steps of cultivating a mammalian cell comprising a deoxyribonucleic acid encoding the multivalent, bispecific antibody, and recovering the multivalent, bispecific antibody from the cell or the cultivation medium, wherein the deoxyribonucleic acid encoding the multivalent, bispecific antibody is stably integrated into the genome of the mammalian cell and comprises in 5?- to 3?-direction a first expression cassette encoding the first heavy chain, a second expression cassette encoding the first light chain, a third expression cassette encoding the first light chain, a fourth expression cassette encoding the second heavy chain, a fifth expression cassette encoding the first light chain, and a sixth expression cassette encoding the first light chain, wherein the first heavy chain comprises from N- to C-terminus a first heavy chain variable domain, a CH1 domain, a first heavy chain variable domain, a CH1 domain, a hinge regi

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: 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.