The invention relates to a tetrameric polypeptide comprising a first polypeptide chain comprising a first VL antigen binding domain and a first CL constant domain, a second polypeptide chain comprising a first VH antigen binding domain, a first CH1 constant domain, a first CH2 constant domain and a first CH3 constant domain, a first ligand binding to a HER2 D4 epitope linked to the N-terminus of said first VL antigen binding domain or said first VH antigen binding domain by a first interdomain amino acid linker, a third polypeptide chain comprising a second VL antigen binding domain and a second CL constant domain, a fourth polypeptide chain comprising a second VH antigen binding domain, a second CH1 constant domain, a second CH2 constant domain and a second CH3 constant domain and a third ligand binding to a HER2 D4 epitope linked to the N-terminus of said second VL antigen binding domain or said second VH antigen binding domain by a second interdomain amino acid linker, wherein the VL antigen binding domains and the VH antigen binding domains together constitute a second ligand and a fourth ligand binding to a HER2 D1 epitope. The invention further relates to the tetrameric polypeptide for use in a method for the prevention or treatment of a malignant neoplastic disease, an isolated nucleic acid and a host cell for expression of the polypeptide and a method for obtaining the polypeptide.

The present invention provides for engineered molecular opsonins that may be used to bind biological pathogens or identify subclasses or specific pathogen species for use in devices and systems for treatment and diagnosis of patients with infectious diseases, blood-borne infections or sepsis. An aspect of the invention provides for mannose-binding lectin (MBL), which is an abundant natural serum protein that is part of the innate immune system. The ability of this protein lectin to bind to surface molecules on virtually all classes of biopathogens (viruses, bacteria, fungi, protozoans) make engineered forms of MBL extremely useful in diagnosing and treating infectious diseases and sepsis.

Herein is reported a method for preparing a recombinant host cell library expressing bispecific antibodies using targeted integration into a host cell comprising an exogenous nucleotide sequence integrated at a site within a locus of the genome of the host 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 whereby the host cell is transfected with a library of first vectors each comprising two recombination recognition sequences matching the first and the third recombination recognition sequence on the integrated exogenous nucleotide sequence and flanking two exogenous nucleotide sequences and at least one second selection marker, and a library of second vectors each comprising two recombination recognition sequences matching the second and the third recombination recognition sequence on the integrated exogenous nucleotide sequence and flanking at least two further exogenous nucleotide sequences; wherein one of the four exogenous nucleotide sequences encode a first light chain, one a second light chain, one a first heavy chain and one a second heavy chain of the bispecific antibody.

The present invention relates to antigen-binding molecules binding to CD3 and CD137 (4-1BB); compositions comprising the antigen-binding molecule; and methods of using the same. The present invention provides antigen-binding molecules comprising: an antibody variable region that is capable of binding to CD3 and CD137 (4-1BB), but does not bind to CD3 and CD137 at the same time; and a variable region binding to a third antigen different from CD3 and CD137. Such antigen binding molecules exhibit enhanced T-cell dependent cytotoxity activity induced by these antigen-binding molecules through binding to the three different antigens.

The present invention relates, in part, to agents that bind CD8 and their use as therapeutic and diagnostic agents. The present invention further relates to pharmaceutical compositions comprising the CD8 binding agents and their use in the treatment of various diseases, including, for example, cancers.

The present invention generally relates to novel bispecific antigen binding molecules for T cell activation and re-direction to specific target cells. In addition, the present invention relates to polynucleotides encoding such bispecific antigen binding molecules, and vectors and host cells comprising such polynucleotides. The invention further relates to methods for producing the bispecific antigen binding molecules of the invention, and to methods of using these bispecific antigen binding molecules in the treatment of disease.

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- to C-terminus a first heavy chain variable domain, a CH1 domain, a hinge region, a CH2 domain and a CH3 domain, the second heavy chain comprises from N- to C-terminus the first light chain variable domain, a CH1 domain, a hinge region, a CH2 domain and a CH3 domain, the first light chain comprises from N- to C-terminus a second heavy chain variable domain and a CL domain, and the second light chain comprises from N- to C-terminus a second light chain variable domain and a CL domain, wherein the first heavy chain variable domain and the second light chain variable domain form a first binding site and the second heavy chain variable domain and the first light chain variable domain form a second binding site.

The present invention provides efficient methods based on alteration of the protein A-binding ability, for producing or purifying multispecific antibodies having the activity of binding to two or more types of antigens to high purity through a protein A-based purification step alone. The methods of the present invention for producing or purifying multispecific antibodies which feature altering amino acid residues of antibody heavy chain constant region and/or variable region. Multispecific antibodies with an altered protein A-binding ability, which exhibit plasma retention comparable or longer than that of human IgG1, can be efficiently prepared in high purity by introducing amino acid alterations of the present invention into antibodies.

The present invention provides compositions for treating canine atopic dermatitis that comprise a caninized anti-proliferative antibody, a caninized anti-pruritic antibody, and a caninized anti-inflammatory antibody. The present invention further relates to compositions that comprise a bispecific antibody that comprises a caninized anti -pruritic antibody and a caninized anti-proliferative antibody or alternatively a bispecific antibody comprising a caninized anti-pruritic antibody and a caninized anti-inflammatory antibody.

The present invention relates to combination therapies employing anti-CD20/anti-CD3 bispecific antibodies and 4-1BB (CD137) agonists, in particular 4-1BBL trimer containing antigen binding molecules, the use of these combination therapies for the treatment of cancer and methods of using the combination therapies.