WO2013157953A1 - Methods and means for the production of ig-like molecules - Google Patents

Methods and means for the production of ig-like molecules Download PDF

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Publication number
WO2013157953A1
WO2013157953A1 PCT/NL2013/050293 NL2013050293W WO2013157953A1 WO 2013157953 A1 WO2013157953 A1 WO 2013157953A1 NL 2013050293 W NL2013050293 W NL 2013050293W WO 2013157953 A1 WO2013157953 A1 WO 2013157953A1
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domain
molecules
different
molecule
nucleic acid
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PCT/NL2013/050293
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English (en)
French (fr)
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Cornelis Adriaan De Kruif
Linda Johanna Aleida HENDRIKS
Ton Logtenberg
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Merus BV
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Merus BV
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Priority to JP2015506927A priority Critical patent/JP6272299B2/ja
Priority to EP13720614.0A priority patent/EP2838917B1/en
Priority to MX2014012700A priority patent/MX360109B/es
Priority to SG11201406770WA priority patent/SG11201406770WA/en
Priority to NZ630551A priority patent/NZ630551A/en
Priority to EP19177410.8A priority patent/EP3594232A1/en
Priority to SM20190493T priority patent/SMT201900493T1/it
Priority to HRP20191446 priority patent/HRP20191446T1/hr
Priority to CA2871068A priority patent/CA2871068C/en
Priority to CN202111365429.7A priority patent/CN114163530B/zh
Priority to ES13720614T priority patent/ES2743399T3/es
Priority to SI201331553T priority patent/SI2838917T1/sl
Priority to RSP20191034 priority patent/RS59263B1/sr
Priority to KR1020147032231A priority patent/KR102104601B1/ko
Priority to US14/395,325 priority patent/US20150196637A1/en
Application filed by Merus BV filed Critical Merus BV
Priority to CN202510431296.0A priority patent/CN120383672A/zh
Priority to EA201491909A priority patent/EA035344B1/ru
Priority to CN201380032190.9A priority patent/CN104520319B/zh
Priority to PL13720614T priority patent/PL2838917T3/pl
Priority to AU2013249985A priority patent/AU2013249985B2/en
Priority to KR1020207011173A priority patent/KR102382304B1/ko
Priority to HK15106743.0A priority patent/HK1206041B/en
Priority to LTEP13720614.0T priority patent/LT2838917T/lt
Priority to DK13720614.0T priority patent/DK2838917T3/da
Publication of WO2013157953A1 publication Critical patent/WO2013157953A1/en
Priority to MX2024012548A priority patent/MX2024012548A/es
Priority to MX2024012547A priority patent/MX2024012547A/es
Anticipated expiration legal-status Critical
Priority to ZA2014/07835A priority patent/ZA201407835B/en
Priority to AU2018201325A priority patent/AU2018201325B2/en
Priority to CY20191100914T priority patent/CY1122118T1/el
Ceased legal-status Critical Current

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    • C12P21/00—Preparation of peptides or proteins
    • C12P21/005—Glycopeptides, glycoproteins
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    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/283—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against Fc-receptors, e.g. CD16, CD32, CD64
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    • C07K16/2896—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against molecules with a "CD"-designation, not provided for elsewhere
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Definitions

  • the invention relates to the fields of molecular biology, medicine and biological therapeutics. It particularly relates to the field of therapeutic antibodies for the treatment of various diseases.
  • Monoclonal antibodies bind to a single specific area, or epitope, of an antigen and, for use in therapy, are often selected for a desirable functional property such as for example killing of tumor cells, blocking of receptor-ligand interactions or virus neutralization.
  • FDA approved monoclonal antibodies which are typically produced at large quantities and their biophysical and biochemical characteristics can be analyzed in great detail to ensure batch-to-batch consistency, which facilitates regulatory acceptability.
  • monoclonal antibodies have several disadvantages, some of which relate to their monospecific nature and the complexity of diseases. Diseases processes are often multifactorial in nature, and involve redundant or synergistic action of disease mediators or up-regulation of different receptors, including crosstalk between their signaling networks.
  • monoclonal antibodies can only interfere with a single step within the complex disease processes which often does not have an optimal effect.
  • targeting a single epitope on a single cellular or soluble protein or pathogen often will not suffice to efficiently treat disease because the target epitope may no longer be available for the monoclonal antibody to bind to and exert the desired effect.
  • tumor cells often escape from monoclonal antibody therapy by down- regulation, mutation or shielding of the target epitope present on a growth factor receptor. By activating alternative receptors and/or their li ands, tumor cells than may exploit a different path leading to continued growth and metastasis.
  • Monoclonal antibodies that bind to a single epitope often do not recruit the full spectrum of effector mechanisms evoked by polyclonal antibodies, including, amongst other things, opsonization (enhancing phagocytosis of antigens), steric hindrance (antigens coated with antibodies are prevented from attaching to host cells or mucosal surfaces), toxin neutralization, agglutination or precipitation (antibodies binding several soluble antigens cause aggregation and subsequent clearance), activation of complement and antibody- dependent cellular cytotoxicity (antibodies enable the killing of target cells by natural killer cells and neutrophils).
  • Polyclonal antibodies for therapeutic applications may be obtained from pooled human serum.
  • serum-derived therapeutic polyclonal antibodies may for example be used to treat or prevent infections caused by viruses such as the rabies virus, cytomegalovirus and respiratory syncytial virus, to neutralize toxins such as tetanus toxin and botulinum toxin or to prevent Rhesus D allograft immunization.
  • viruses such as the rabies virus, cytomegalovirus and respiratory syncytial virus
  • toxins such as tetanus toxin and botulinum toxin or to prevent Rhesus D allograft immunization.
  • a more widespread use of serum-derived polyclonal antibody preparations has been prevented by the fact that source plasma is only available for a limited range of targets such as infectious diseases and toxins.
  • the products are highly dependent on donor blood availability, both in terms of quantity and suitability, resulting in considerable variation between batches.
  • combinations of 2 monoclonal antibodies may have additive or synergistic effects and recruit effector mechanisms that are not associated with either antibody alone.
  • mixtures of 2 monoclonal antibodies against the EGFR or HER2 were shown to more potently kill tumor cells based on a combination of activities including enhanced receptor internalization, improved blockade of signalling pathways downstream of the receptors as well as enhanced immune effector-mediated cytotoxicity.
  • the component antibodies may be produced separately and combined at the protein level.
  • a drawback of this approach is the staggering cost of developing the 2 antibodies individually in clinical trials and (partially) repeating that process with the combination. This would lead to unacceptable cost of treatments based on antibody combinations.
  • the 2 recombinant cell lines producing the component monoclonal antibodies may be mixed in a fermentor and the resultant mixture of antibodies may be purified as a single preparation (WO 2004/061104).
  • a drawback of this approach is the poor control over the composition and hence reproducibility of the resulting recombinant polyclonal antibody preparation, especially when considering that such compositions may change over time as the cells are being cultured.
  • bispecific antibodies have emerged as an alternative to the use of combinations of 2 antibodies. Whereas a combination of 2 antibodies represents a mixture of 2 different immunoglobulin molecules that bind to different epitopes on the same or different targets, in a bispecific antibody this is achieved through a single immunoglobulin molecule. By binding to 2 epitopes on the same or different targets, bispecific antibodies may have similar effects as compared to a combination of 2 antibodies binding to the same epitopes. Furthermore, since bispecific antibodies of the IgG format combine 2 different monovalent binding regions in a single molecule and mixtures of 2 IgG antibodies combine 2 different bivalent binding molecules in a single preparation, different effects of these formats have been observed as well.
  • bispecific antibodies based on the IgG format consisting of 2 heavy and two light chains have been produced by a variety of methods. For instance, bispecific antibodies may be produced by fusing two antibody-secreting cell lines to create a new cell line or by expressing two antibodies in a single cell using recombinant DNA technology.
  • the invention provides methods and means for improved and/or alternative technologies for producing biological therapeutics in the form of mixtures or bispecific approaches for targeting multiple disease-modifying molecules, as well as products and uses resulting from these methods and means.
  • CH3-CH3 interaction is the primary driver for Fc dimerization (Ellerson JR., et al., J. Immunol 1976 (116) 510-517; Deisenhofer J. biochemistry 1981 (20) 2361-2370). It is furthermore well-known that when two CH3 domains interact with each other they meet in a protein-protein interface which comprises "contact" residues (also called contact amino acids, interface residues or interface amino acids). Contact amino acids of a first CH3 domain interact with one or more contact amino acids of a second CH3 domain. Contact amino acids are typically within 5.5 A (preferably within 4.5 A) of each other in the three-dimensional structure of an antibody.
  • contact residues from one CH3 domain and contact residues from a different CH3 domain may for instance be via Van der Waals forces, hydrogen bonds, water-mediated hydrogen bonds, salt bridges or other electrostatic forces, attractive interactions between aromatic side chains, disulfide bonds, or other forces known to one skilled in the art. It was previously shown that approximately one-third of the contact amino acid side chains at the human IgGl
  • CH3 domain interface can account for the majority of contributions to domain folding and association. It can further be envisaged that other (neighbouring) amino acid residues may affect the interactions in the protein-protein interface. Approaches to interfere with the dimerization of antibody heavy chains have been employed in the art. Specific engineering in the CH3 domains was applied in order to favour heterodimerization over homodimerization. Examples of such engineering of the CH3-CH3 interface include the introduction of complementary protuberance and cavity mutations, also known as 'knob-into-hole' approaches as described for instance in WO 1998/050431, Ridgeway et al., 1996 and Merchant et al. 1998.
  • the method involves introducing a protuberance at the interface of a first polypeptide and a corresponding cavity in the interface of a second polypeptide, such that the protuberance can be positioned in the cavity so as to promote heteromultimer formation and hinder homomultimer formation.
  • protuberances or “knobs” are constructed by replacing small amino acid side chains from the interface of the first polypeptide with larger side chains (e.g. tyrosine or tryptophan).
  • Compensatory "cavities” or “holes” of identical or similar size to the protuberances are created in the interface of the second polypeptide by replacing large amino acid side chains with smaller ones (e.g. alanine or threonine).
  • the protuberance and cavity can be made by synthetic means such as altering the nucleic acid encoding the polypeptides or by peptide synthesis.
  • the proportion of a bispecific antibody of interest is at best 87% of the mixture of the 2 parental and bispecific antibodies.
  • bispecific antibody has to be purified (separated) from the homodimers and formulated into a pharmaceutically acceptable diluent or excipient. Purification of heterodimers from such mixtures poses a major challenge because of the similarity in physico-chemical properties of the homodimers and heterodimers. It is one object of the present invention to provide methods for producing a bispecific antibody in a single cell clone with a further improved proportion of the bispecific antibody in the mixture. According to the invention, knob-into-hole technology can thus be used as one of the means, alone or together with other means, to achieve said further improved bispecific proportion in a mixture.
  • SEED CH3 heterodimeric Fc technology that supports the design of bispecific and asymmetric fusion proteins by devising strand-exchange engineered domain (SEED) CH3 heterodimers.
  • SEED CH3 heterodimers are derivatives of human IgG and IgA CH3 domains that are composed of alternating segments of human IgA and IgG CH3 sequences which results in pairs of complementary human SEED CH3 heterodimers, the so-called SEED-bodies (Davis JH. Et al., Protein Engineering, Design & Selection 2010(23)195-202; WO2007/110205).
  • bispecific antibody in a single cell with proportions ranging between about 76% and about 96%. It is an object of the present invention to provide methods for producing a bispecific antibody in a single cell with a further improved percentage of desired bispecific antibodies.
  • electrostatic engineering technology can be used as one of the means, alone or together with other means, e.g knob-into-hole approaches, to achieve said further improved percentages of desired (bispecific) antibodies.
  • the present invention provides a method for producing at least two different Ig-like molecules from a single host cell, wherein each of said two Ig-like molecules comprises two CH3 domains that are capable of forming an interface, said method comprising providing in said cell
  • nucleic acid molecules are provided with means for preferential pairing of said 1 st and 2 nd CH3 domain-comprising polypeptides and said 3 rd and 4 th CH3-domain comprising polypeptides, said method further comprising the step of culturing said host cell and allowing for expression of said at least four nucleic acid molecules and harvesting said at least two different Ig-like molecules from the culture.
  • bispecific antibody It is often desired to produce more than one (bispecific) antibody, for instance in order to more efficiently interfere with multiple biological pathways involved in a disease process or with the invasion, replication and/or spreading of a pathogen.
  • a mixture of more than one bispecific antibody is also particularly useful for the treatment of certain diseases.
  • tumor cells use many different strategies to develop resistance during treatment with antibodies or small molecule drugs. Resistance may involve multiple cell surface receptors and soluble molecules and it is considered beneficial to develop antibody-based treatments for cancers that address multiple such disease- and escape-associated molecules simultaneously.
  • a mixture of bispecific antibodies provides an innovative and attractive therapeutic format.
  • such mixtures of bispecific antibodies are produced by a single cell to facilitate a drug development process that is less complicated from a regulatory point of view and cost-effective and feasible from a drug m anufacturing and clinical development point of view .
  • a single cell-based approach it is desirable to use methods that allow controlled and efficient production of the bispecific antibodies, thus reducing or even completely abrogating the need of separating the desired mixture of bispecific IgG molecules from non-desired monospecific IgG molecules.
  • mixtures of monospecific and bispecific antibodies have been produced by a single cell (WO2004/009618), but these mixtures represent complex concoctions of several different bispecific and monospecific antibody species.
  • methods are provided which result in mixtures of (bispecific) antibodies with a proportion of at least 95%, at least 97% or even more than 99% of dimeric IgG molecules, irrespective of the amount of monomeric by-products, see herein below.
  • half molecules monomeric by-products
  • the present invention provides methods for producing a defined mixture of at least two different Ig-like molecules in single cells, instead of a single (bispecific) antibody of interest, wherein the formation of other, undesired dimeric antibody species is diminished or even absent.
  • the resulting mixture is well defined and its composition is controlled by the design of CH3 domain mutants. Furthermore, regulation of expression levels and/or different transfection ratios used for expression affects the composition of the mixture.
  • a first nucleic acid molecule encodes a CH3 domain which preferentially pairs with a CH3 domain encoded by a second nucleic acid molecule
  • a third nucleic acid molecules encodes a CH3 domain which preferentially pairs with a CH3 domain encoded by a fourth nucleic acid molecule.
  • the present invention also provides mixtures of at least two different Ig-like molecules obtainable by the methods of the invention.
  • the term "preferential pairing of said 1 st and 2 nd CH3 domain- comprising polypeptides" means that essentially all the resulting dimers comprising the 1 st CH3 domain-comprising polypeptide and/or the 2 nd CH3 domain-comprising polypeptide will be dimers consisting of one 1 st CH3 domain-comprising polypeptide paired with one 2 nd CH3 domain-comprising polypeptide.
  • the term “preferential pairing of said 1 st and 2 nd CH3 domain- comprising polypeptides” means that essentially all the resulting dimers comprising the 1 st CH3 domain-comprising polypeptide and/or the 2 nd CH3 domain-comprising polypeptide will be dimers consisting of one 1 st CH3 domain-comprising polypeptide paired with one 2 nd CH3 domain-comprising polypeptide.
  • "preferential pairing of said 3 rd and 4 th CH3 domain-comprising polypeptides” means that essentially all of the resulting dimers comprising the 3 rd CH3 domain-comprising polypeptide and/or the 4 th CH3 domain-comprising polypeptide will be dimers consisting of one 3 rd CH3 domain-comprising polypeptide paired with one 4 th CH3 domain-comprising polypeptide.
  • nucleic acid molecules encoding four different (A, B, C, D) CH3 domain-comprising polypeptides are introduced in a single cell, instead of a mixture of 10 different Ig-like dimers (AA, AB, AC, AD, BB,
  • said first CH3- domain comprising polypeptide chain comprises the amino acid substitution T366K
  • said second CH3-domain comprising polypeptide chain comprises the amino acid substitution L351D.
  • Said first CH3-domain comprising polypeptide chain preferably further comprises the amino acid substitution L351K.
  • said second CH3-domain comprising polypeptide chain preferably further comprises an amino acid substitution selected from the group consisting of Y349E, Y349D and L368E, most preferably L368E.
  • said third CH3-domain comprising polypeptide chain comprises the amino acid substitutions E356K and D399K
  • said fourth CH3- domain comprising polypeptide chain comprises the amino acid substitutions K392D and K409D.
  • each of the CH3-domain comprising polypeptide chains preferably further comprises a variable region recognizing a target epitope.
  • the variable regions that are part of the CH3-domain comprising polypeptide chains preferably share a common light chain. In that case only the VHs of the variable regions differ whereas the VL in all variable regions is essentially the same.
  • a method according to the invention which further comprises providing said host cell with a nucleic acid molecule encoding a common light chain.
  • each of said 4 variable regions of the 4 CH3-domain comprising polypeptide chains recognizes a different target epitope.
  • the first nucleic acid molecule encodes a heavy chain that further contains a variable domain with specificity for antigen A
  • the second nucleic acid molecule encodes a heavy chain that further contains a variable domain with specificity for antigen B
  • the third nucleic acid molecule encodes a heavy chain that further contains a variable domain with specificity for antigen C
  • the fourth nucleic acid molecule encodes a heavy chain that further contains a variable domain with specificity for antigen D
  • the ratio of the nucleic acids used in a method according to the invention does not need to be 1: 1: 1: 1 and the ratio of the resulting Ig-like molecules that are expressed does not need to be 1: 1. It is possible to use means known in the art to produce mixtures of antibodies with optimized ratios. For instance, expression levels of nucleic acid molecules and hence the ratios of the resulting Ig-like molecules produced may be regulated by using different genetic elements such as promoters, enhancers and repressors or by controlling the genomic integration site of copy number of the DNA constructs encoding antibodies.
  • Said means for preferential pairing preferably may comprise engineered
  • said means for preferential pairing may be chosen within a certain type of mutations, i.e. all at least 4 nucleic acid molecules encoding CH3-domain comprising polypeptide chains may for example comprise charge mutations as means for preferential pairing.
  • non-engineered wildtype CH3 may in certain instances be used for preferential pairing of two wildtype CH3-domain comprising polypeptide chains.
  • said means for preferential pairing comprise at least one CH3 mutation selected from Table B, as explained elsewhere in this application.
  • One preferred embodiment thus provides a method according to the present invention, wherein all 4 of said nucleic acid molecules are provided with means for preferential pairing of said 1 st and 2 nd CH3 domain- comprising polypeptides and said 3 rd and 4 th CH3-domain comprising polypeptides, wherein said means for preferential pairing of said 1 st and 2 nd CH3 domain- comprising polypeptides are different from those means for preferential pairing of said 3 rd and 4 th CH3-domain comprising polypeptides.
  • One aspect of the present invention provides a method according to the invention, wherein said means for preferential pairing of said 1 st and 2 nd CH3 domain- comprising polypeptides are different from said means for preferential pairing of said 3 rd and 4 th CH3-domain comprising polypeptides.
  • said means for preferential pairing of said 1 st and 2 nd CH3 domain comprising polypeptides are designed such that preferential pairing of the 1 st and 2 nd chain is favoured.
  • the design is such that essentially no interaction between the 1 st and the 3 rd and/or 4 th CH3 domain comprising polypeptide chain will take place.
  • the 3 rd and the 4 th CH3 domain-comprising polypeptides may either be wildtype or may comprise means for preferential pairing that are different from the means for preferential pairing of the 1 st and 2 nd CH3 domains.
  • Current studies have focused on the production of a single bispecific antibody, using for instance the knob-into-hole technology or mutations (reversions) of charged contact amino acids present in CH3 domains.
  • the present invention provides methods for the efficient and controlled production of a well-defined mixture of Ig-like molecules, with a high proportion of bispecifics in the mixture. Even a proportion of (two) bispecifics of at least 95%, at least 97% or more is obtained in a system where two bispecifics are desired. This means that only at most 5%, at most 3% or less monospecific bivalent by-products are obtained. Of note, the amount of monomeric by-products, i.e. half molecules, is less important since these half-molecules are easily separated from dimers using their size difference.
  • variable regions of the 1 st and the 2 nd CH3- domain comprising polypeptide chains recognize different target epitopes, whereas the variable regions of the 3 rd and the 4 th CH3-domain comprising polypeptide chains recognize the same target epitopes. This will result in the predominant production of one kind of bispecific Ig-like molecule and one kind of monospecific Ig-like molecule.
  • variable regions of the 1 st and the 2 nd CH3-domain comprising polypeptide chains recognize different target epitopes and if the variable regions of the 3 rd and the 4 th CH3-domain comprising polypeptide chains both recognize the same target epitope which is different from the target epitopes recognized by the 1 st and the 2 nd CH3-domains, a mixture of Ig-like molecules having specificity for AB or CC will be formed.
  • the target epitope recognized by the variable regions of the 3rd and 4 th CH3 domain comprising polypeptide chain is the same, but different from the target epitope recognized by the variable region of the 1 st or the 2 nd CH3-domain comprising polypeptide chain.
  • variable regions of the 1 st and the 2 nd CH3-domain comprising polypeptide chains recognize different target epitopes and when the variable regions of the 3 rd and the 4 th CH3-domain comprising polypeptide chains both recognize the same epitope as the 1 st or the 2 nd CH3-domain comprising polypeptide chains, a mixture of Ig-like molecules having specificity for AB and AA, or AB and BB will be formed.
  • a method according to the invention wherein the target epitope recognized by the variable regions of the 3rd and 4 th CH3 domain comprising polypeptide chain is the same as the target epitope recognized by the variable region of the 1 st or the 2 nd CH3-domain comprising polypeptide chain is therefore also herewith provided. It is another object of the present invention to provide means and methods for producing defined mixtures of bispecific antibodies and monospecific antibodies in a single cell culture.
  • a non-limiting example of such well-defined mixture is a mixture of bispecific antibodies with specificity AB and monospecific antibodies with specificity AA.
  • Another example is a mixture of bispecific antibodies with specificity AB and monospecific antibodies with specificity BB.
  • Yet another example is a mixture of bispecific antibodies with specificity AB and monospecific antibodies with specificity CC.
  • means and methods are provided which yield mixtures of antibodies of interest with at least 90%, more preferably at least 95% and most preferably at least 97% or even more than 99% of desired antibodies.
  • variable regions of the 1 st and the 2 nd CH3-domain comprising polypeptide chains recognize the same target epitope, whereas the variable regions of the 3 rd and the 4 th CH3-domain comprising polypeptide chains recognize a second target epitope which differs from the target epitope recognized by said 1 st and 2 nd variable regions.
  • This will result in the predominant production of monospecific Ig-like molecules having either specificity for AA or specificity for BB.
  • the formation of bispecific Ig-like molecules is diminished or even avoided.
  • a method according to the present invention is suitable for the production of any desired mixture of bispecific and/or monospecific Ig-like molecules.
  • further nucleic acid molecules for instance encoding a 5 th and a 6 th (and 7 th and 8 th and so forth) CH3 domain-comprising polypeptide, in order to produce defined mixtures comprising more than two different Ig-like molecules.
  • at least two CH3 domains are used that comprise at least one combination of mutations provided by the present invention. Through these mutations novel specific interactions are formed between two CH3 domains.
  • Ig-like molecule means a proteinaceous molecule that possesses at least one immunoglobulin (Ig) domain.
  • Said Ig-like molecule comprises a sequence comprising the function of at least an immunoglobulin CH3 domain, preferably the sequence comprises an IgGl CH3 domain.
  • Proteinaceous molecules that possess at least a CH3 domain can be further equipped with specific binding moieties.
  • binding moieties that can be engineered to the CH3-domain comprising proteinaceous molecules can be any binding agent, including, but not limited to, single chain Fvs, single chain or Tandem diabodies (TandAb®), VHHs, Anticalins®, Nanobodies®, a BiTE®, a Fab, ankyrin repeat proteins or DARPINs®, Avimers®, a DART, a TCR-like antibody, Adnectins®, Affilins®, Trans-bodies®, Affibodies®, a TrimerX®, MicroProteins, Fynomers®, Centyrins® or a KALBITOR®.
  • TandAb® Tandem diabodies
  • the binding moieties are antibody variable regions (i.e. VH/VL combinations).
  • VH/VL combinations antibody variable regions that are part of the CH3-domain comprising polypeptide chains preferably share a common light chain. In that case, only the VHs of the variable regions differ whereas the VL in all variable regions is essentially the same.
  • CH3 domains of the present invention can be engineered to the CH3 domains of the present invention, including cytokines, hormones, soluble ligands, receptors and/or peptides.
  • said Ig-like molecule comprises a full length Fc backbone.
  • the Ig-like molecules are antibodies.
  • the variable regions of these antibodies preferably share a common light chain, but they may differ in their VH regions.
  • the term 'antibody' as used herein means a
  • Antibodies are known in the art and include several isotypes, such as IgGl, IgG2, IgG3, IgG4, IgA, IgD, IgE, and IgM.
  • An antibody according to the invention may be any of these isotypes, or a functional derivative and/or fragment of these.
  • Ig-like molecules are produced that are antibodies of the IgG isotype because IgG antibodies e.g. have a longer half life as compared to antibodies of other isotypes.
  • Antibodies produced with methods according to the present invention can have sequences of any origin, including murine and human sequences.
  • Antibodies can consist of sequences from one origin only, such as fully human antibodies, or they can have sequences of more than one origin, resulting for instance in chimeric or humanized antibodies.
  • Antibodies for therapeutic use are preferably as close to natural antibodies of the subject to be treated as possible (for instance human antibodies for human subjects).
  • Antibody binding can be expressed in terms of specificity and affinity. The specificity determines which antigen or epitope thereof is bound by the binding domain.
  • the affinity is a measure for the strength of binding to a particular antigen or epitope.
  • Specific binding is defined as binding with affinities (KD) of at least lxl0 5 M, more preferably lxl0 "7 M, more preferably higher than 1x10 " 9 M.
  • KD affinities
  • monoclonal antibodies for therapeutic applications have affinities of up to lxl0 10 M or even higher.
  • the term 'antigen' as used herein means a substance or molecule that, when introduced into the body, triggers the production of an antibody by the immune system.
  • An antigen among others, may be derived from pathogenic organisms, tumor cells or other aberrant cells, from haptens, or even from self structures. At the molecular level, an antigen is characterized by its ability to be bound by the antigen-binding site of an antibody.
  • An antigen comprises at least one, but often more, epitopes.
  • the term 'epitope' as used herein means a part of an antigen that is recognized by the immune system, specifically by antibodies, B cells, or T cells. Although epitopes are usually thought to be derived from non-self proteins, sequences derived from the host that can be recognized are also classified as epitopes.
  • the term 'CH3 domain' is well known in the art.
  • the IgG structure has four chains, two light and two heavy chains; each light chain has two domains, the variable and the constant light chain (VL and CL) and each heavy chain has four domains, the variable heavy chain (VH) and three constant heavy chain domains (CHI, CH2, CH3).
  • the CH2 and CH3 domain region of the heavy chain is called Fc (Fragment crystallizable) portion, Fc fragment, Fc backbone or simply Fc.
  • the IgG molecule is a heterotetramer having two heavy chains that are held together by disulfide bonds (-S- S-) at the hinge region and two light chains. The heavy chains dimerize through interactions at the CH3-CH3 domain interface and through interactions at the hinge region.
  • the number of hinge disulfide bonds varies among the immunoglobulin subclasses (Papadea and Check 1989).
  • the Fc fragment of an immunoglobulin molecule is a dimer of the two C-terminal constant regions, i.e. CH2 and CH3 domains, of the heavy chain.
  • CH2 and CH3 domains a dimer of the two C-terminal constant regions, i.e. CH2 and CH3 domains
  • CH3 domains direct the association of antibody heavy chains, and it is known that the interface between CH3 domains contains more than 20 contact residues from each chain that play a role in the CH3-CH3 interaction (Deisenhofer J., Biochemistry 1981(20)2361-2370; Miller S., J. Mol. Biol. 1990(216)965-973; Padlan, Advances in Protein Chemistry 1996 (49) 57- 133).
  • the CH3 variants of the present invention can thus be used in association with other antibody domains to generate full length antibodies that are either bispecific or monospecific.
  • the specificity of the antibody as defined by the VH/VL combinations typically does not affect the heavy chain dimerization behaviour that is driven by the CH3 domains.
  • 'contact residue', 'contact amino acid', 'interface residue' and 'interface amino acid' typically refers to any amino acid residue present in the CH3 domain that can be involved in interdomain contacts, as can be calculated by technologies known in the art, including calculating solvent accessible surface area (ASA) of the CH3 domain residues in the presence and absence of the second chain (Lee and Richards J. Mol. Biol.1971(55)379) where residues that show difference (> 1A 2 ) in ASA between the two calculations are identified as contact residues.
  • ASA solvent accessible surface area
  • Contact residues that have been identified include residues at positions 347, 349, 350, 351, 352, 353, 354, 355, 356, 357, 360, 364, 366, 368, 370, 390, 392, 394, 395, 397, 399, 400, 405, 407, 409, 439 according to the EU numbering system (Table A).
  • Contact residues within the CH3-CH3 interface can either be amino acids that are charged, or amino acid residues that are neutral.
  • the term 'charged amino acid residue' or 'charged residue' as used herein means amino acid residues with electrically charged side chains. These can either be positively charged side chains, such as present in arginine (Arg, R), histidine (His, H) and lysine (Lys, K) or can be negatively charged side chains, such as present in aspartic acid (Asp, D) and glutamic acid (Glu, E).
  • the term 'neutral amino acid residue' or neutral residue as used herein refers to all other amino acids that do not carry electrically charged side chains.
  • neutral residues include serine (Ser, S), threonine (Thr, T), asparagine (Asn, N), glutamine (GLu, Q), Cysteine (Cys, C), glycine (Gly, G), proline (Pro, P), alanine (Ala, A), valine (Val, V), isoleucine (He, I), leucine (Leu, L), methionine (Met, M), phenylalanine (Phe, F), tyrosine (Tyr, Y), and tryptophan (Trp, T).
  • 'CH3-CH3 domain interface' refers to the association between two CH3 domains of separate CH3-domain comprising polypeptides that is a result of interacting amino acid residues, i.e. at least one interaction between an amino acid of a first CH3 domain and an amino acid of a second CH3 domain.
  • Such interaction is for instance via Van der Waals forces, hydrogen bonds, water-mediated hydrogen bonds, salt bridges or other electrostatic forces, attractive interactions between aromatic side chains, the formation of disulfide bonds, or other forces known to one skilled in the art.
  • said means for preferential pairing of the first and second CH3 domain-comprising polypeptides and said third and fourth CH3 domain-comprising polypeptide can be any means known in the art.
  • at least one nucleic acid molecule encodes a CH3 domain which contains at a contact residue position a large amino acid residue (i.e. a "knob” or "protuberance”) such as for instance R, F, Y, W, I or L
  • at least one other nucleic acid molecule encodes a CH3 domain which contains at a complementary contact residue position a small amino acid residue (i.e. a "hole” or "cavity”) such as for instance G, A, S, T or V.
  • At least one nucleic acid molecule encodes a CH3 domain which contains at a contact residue position that is naturally charged, i.e. a naturally occurring K, H, R, D or E, an amino acid that now carries the opposite charge as compared to wildtype
  • at least one other nucleic acid molecule encodes a CH3 domain which contains at a complementary contact residue position that is naturally charged, an amino acid that now carries the opposite charge as compared to wildtype.
  • the resulting engineered CH3 domains will preferentially pair with each other due to the opposite charges of said contact amino acids, whereas pairing of identical CH3 domains will be
  • CH3 mutations as described in EPO 1870459, WO 2009/089004, Gunasekaran et al (2010), are used.
  • the means for preferential pairing of said 1 st and 2 nd CH3 domain- comprising polypeptides are "knob" and "hole” amino acid residues and the means for preferential pairing of said 3 th and 4 th CH3 domain-comprising polypeptides are charge-engineered amino acids.
  • both said means for preferential pairing of said 1 st and 2 nd CH3 domain-comprising polypeptides and said 3 th and 4 th CH3 domain-comprising polypeptides are charge-engineered amino acids.
  • different amino acid residues are engineered for preferential pairing of said 1 st and 2 nd CH3 domain-comprising polypeptides as compared to the amino acid residues that are engineered for preferential pairing of said 3 th and 4 th CH3 domain- comprising polypeptides.
  • at least a first and a second nucleic acid molecule encode CH3 domains with novel mutations as provided by the present invention.
  • the present invention provides novel CH3 mutations which enable the production of certain bispecific Ig-like molecules of interest without a significant amount of undesired (dimeric) by-products.
  • the present invention also provides novel CH3 mutations which enable the production of certain monospecific Ig-like molecules of interest without a significant amount of undesired (dimeric) by-products.
  • novel CH3 mutations which enable the production of certain monospecific Ig-like molecules of interest without a significant amount of undesired (dimeric) by-products.
  • the use of at least one of these CH3 mutations according to the present invention is, therefore, preferred.
  • polypeptide', 'polypeptide molecule' or 'polypeptide chain' refers to a chain of amino acids that are covalently joined together through peptide bonds. Proteins are typically made up of one or more polypeptide molecules. One end of every polypeptide, called the amino terminal or N-terminal, has a free amino group. The other end, with its free carboxyl group, is called the carboxyl terminal or C- terminal. Polypeptides according to the present invention may have gone through post-translational modification processes and may e.g. be glycosylated.
  • the CH3 domain-comprising polypeptide chains of the present invention thus refer to polypeptide chains that at least encompass an Ig CH3 domain and that may have gone through post-translational modification processes.
  • nucleic acid molecule as used herein is defined as a molecule comprising a chain of nucleotides, more preferably DNA and/or RNA. In one embodiment, double- stranded RNA is used. In other embodiments a nucleic acid molecule of the invention comprises other kinds of nucleic acid structures such as for instance a DNA/RNA helix, peptide nucleic acid (PNA), locked nucleic acid (LNA) and/or a ribozyme. Hence, the term “nucleic acid molecule” also encompasses a chain comprising non-natural nucleotides, modified nucleotides and/or non-nucleotide building blocks which exhibit the same function as natural nucleotides.
  • the present invention further provides a method for making a host cell for production of at least two different Ig-like molecules, the method comprising the step of introducing into said host cell nucleic acid sequences encoding at least a first, a second, a third and a fourth CH3-domain comprising polypeptide chain, wherein at least two of said nucleic acid sequences are provided with means for preferential pairing of said first and second CH3-domain comprising polypeptides and said third and fourth CH3-domain comprising polypeptides, wherein said nucleic acid sequences are introduced consecutively or concomitantly.
  • Said methods for making said host cells preferably further comprise the step of introducing into said host cell a nucleic acid sequence encoding a common light chain.
  • Also provided herein is a recombinant host cell comprising nucleic acid sequences encoding at least a first, a second, a third and a fourth CH3-domain comprising polypeptide chain, wherein at least two of said nucleic acid molecules are provided with means for preferential pairing of said first and second CH3-domain comprising polypeptides and said third and fourth CH3-domain comprising polypeptides.
  • the invention furthermore provides a recombinant host cell comprising nucleic acid sequences encoding at least a first and a second CH3-domain comprising polypeptide chain, wherein said first CH3 domain-comprising polypeptide chain comprises at least one substitution of a neutral amino acid residue by a positively charged amino acid residue and wherein said second CH3 domain-comprising polypeptide chain comprises at least one substitution of a neutral amino acid residue by a negatively char ⁇ amino acid residue.
  • a recombinant host cell according to the invention preferably further comprises nucleic acid sequence encoding a common light chain.
  • a "host cell” according to the invention may be any host cell capable of expressing recombinant DNA molecules, including bacteria such as for instance Escherichia (e.g. E. coli , Enterobacter, Salmonalla, Bacillus, Pseudomonas, Streptomyces, yeasts such as S. cerevisiae, K. lactis, P.
  • Escherichia e.g. E. coli , Enterobacter, Salmonalla, Bacillus, Pseudomonas, Streptomyces
  • yeasts such as S. cerevisiae, K. lactis, P.
  • filamentous fungi such as Neurospora, Aspergillus oryzae, Aspergillus nidulans and Aspergillus niger, insect cells such as Spodoptera frugiperda SF-9 or SF-21 cells, and preferably mammalian cells such as Chinese hamster ovary (CHO) cells, BHK cells, mouse cells including SP2/0 cells and NS-0 myeloma cells, primate cells such as COS and Vero cells, MDCK cells, BRL 3A cells, hybridomas, tumor-cells, immortalized primary cells, human cells such as W138, HepG2, HeLa, HEK293, HT1080 or embryonic retina cells such as PER. C6, and the like. Often, the expression system of choice will involve a
  • a human cell line preferably PER.C6
  • PER.C6 can advantageously be used to obtain antibodies with a completely human glycosylation pattern.
  • the conditions for growing or multiplying cells see e. g. Tissue Culture, Academic Press, Kruse and Paterson, editors (1973)
  • the conditions for expression of the recombinant product may differ somewhat, and optimization of the process is usually performed to increase the product proportions and/or growth of the cells with respect to each other, according to methods generally known to the person skilled in the art.
  • principles, protocols, and practical techniques for maximizing the productivity of mammalian cell cultures can be found in Mammalian Cell
  • the nucleic acid molecules encoding the light and heavy chains may be present as extrachromosomal copies and/or stably integrated into the chromosome of the host cell, the latter is preferred. It is a further aspect of the present invention to provide a culture of recombinant host cells according to the invention, or a culture of recombinant host cells obtainable or obtained by a method according to the invention, said culture either producing at least two different Ig-like molecules or a heterodimeric Ig-like molecule.
  • nucleic acid sequences encoding the CH3 domain-comprising polypeptides it is well known to those skilled in the art that sequences capable of driving such expression can be functionally linked to the nucleic acid sequences encoding the CH3 domain-comprising polypeptides.
  • Functionally linked is meant to describe that the nucleic acid sequences encoding the CH3 domain-comprising polypeptides or precursors thereof is linked to the sequences capable of driving expression such that these sequences can drive expression of the CH3 domain- comprising polypeptides or precursors thereof.
  • Useful expression vectors are available in the art, e.g. the pcDNA vector series of Invitrogen.
  • Sequences driving expression may include promoters, enhancers and the like, and combinations thereof. These should be capable of functioning in the host cell, thereby driving expression of the nucleic acid sequences that are functionally linked to them. Promoters can be constitutive or regulated, and can be obtained from various sources, including viruses, prokaryotic, or eukaryotic sources, or artificially designed.
  • Expression of nucleic acids of interest may be from the natural promoter or derivative thereof or from an entirely heterologous promoter.
  • Some well-known and much used promoters for expression in eukaryotic cells comprise promoters derived from viruses, such as adenovirus, e.g. the E1A promoter, promoters derived from cytomegalovirus (CMV), such as the CMV immediate early (IE) promoter, promoters derived from Simian Virus 40 (SV40), and the like.
  • viruses such as adenovirus, e.g. the E1A promoter, promoters derived from cytomegalovirus (CMV), such as the CMV immediate early (IE) promoter, promoters derived from Simian Virus 40 (SV40), and the like.
  • CMV cytomegalovirus
  • IE CMV immediate early
  • SV40 Simian Virus 40
  • Suitable promoters can also be derived from eukaryotic cells, such as methallothionein (MT) promoters, elongation factor la (EF- la) promoter, actin promoter, an immunoglobulin promoter, heat shock promoters, and the like.
  • Any promoter or enhancer/promoter capable of driving expression of the sequence of interest in the host cell is suitable in the invention.
  • the sequence capable of driving expression comprises a region from a CMV promoter, preferably the region comprising nucleotides -735 to +95 of the CMV immediate early gene enhancer/promoter.
  • the expression sequences used in the invention may suitably be combined with elements that can stabilize or enhance expression, such as insulators, matrix attachment regions, STAR elements (WO 03/004704), and the like. This may enhance the stability and/or levels of expression.
  • Protein production in recombinant host cells has been extensively described, e.g. in Current Protocols in Protein Science, 1995, Coligan JE, Dunn BM, Ploegh HL, Speicher DW, Wingfield PT, ISBN 0-471- 11184-8; Bendig, 1988.
  • Culturing a cell is done to enable it to metabolize, and/or grow and/or divide and/or produce recombinant proteins of interest. This can be accomplished by methods well known to persons skilled in the art, and includes but is not limited to providing nutrients for the cell.
  • the methods comprise growth adhering to surfaces, growth in suspension, or combinations thereof.
  • Several culturing conditions can be optimized by methods well known in the art to optimize protein production yields.
  • Culturing can be done for instance in dishes, roller bottles or in bioreactors, using batch, fed-batch, continuous systems, hollow fiber, and the like.
  • continuous production of recombinant proteins through cell culture it is preferred in the art to have cells capable of growing in suspension, and it is preferred to have cells capable of being cultured in the absence of animal- or human-derived serum or animal- or human-derived serum components.
  • purification is easier and safety is enhanced due to the absence of additional animal or human proteins derived from the culture medium, while the system is also very reliable as synthetic media are the best in reproducibility.
  • Ig-like molecules are expressed in host cells and are harvested from the cells or, preferably, from the cell culture medium by methods that are generally known to the person skilled in the art. After harvesting, these Ig-like molecules may be purified by using methods known in the art. Such methods may include precipitation,
  • protein A or protein G affinity chromatography can be suitably used (see e.g. US patents 4,801,687 and 5, 151,504).
  • Ig-like molecules, and/or mixtures thereof, produced with methods according to the present invention preferably have a common light chain.
  • a method according to the invention further comprising providing said host cell with a nucleic acid molecule encoding a common light chain.
  • This is a light chain that is capable of pairing with at least two different heavy chains, thereby forming functional antigen binding domains.
  • a functional antigen binding domain is capable of specifically binding to an antigen.
  • a common light chain is used that is capable of pairing with all heavy chains produced with a method according to the invention, thereby forming functional antigen binding domains, so that mispairing of unmatched heavy and light chains is avoided.
  • only common light chains with one identical amino acid sequence are used.
  • “common” also refers to functional equivalents of the light chain of which the amino acid sequence is not identical.
  • Many variants of said light chain exist wherein mutations (deletions, substitutions, additions) are present that do not materially influence the formation of functional binding regions. Such variants are thus also capable of binding different heavy chains and forming functional antigen binding domains.
  • the term 'common light chain' as used herein thus refers to light chains which may be identical or have some amino acid sequence differences while retaining the binding specificity of the resulting antibody after pairing with a heavy chain. It is for instance possible to prepare or find light chains that are not identical but still functionally equivalent, e.g.
  • a common light chain is used in the present invention which is a germline- like light chain, more preferably a germline light chain, preferably a rearranged germline human kappa light chain, most preferably either the rearranged germline human kappa light chain IgVKl-39/JK or IGVK3-20/JK.
  • a common light chain is used in the present invention which is a germline- like light chain, more preferably a germline light chain, preferably a rearranged germline human kappa light chain, most preferably either the rearranged germline human kappa light chain IgVKl-39/JK or IGVK3-20/JK.
  • the skilled person may select, as an alternative to using a common light chain and to avoid mispairing of unmatched heavy and light chains, means for forced pairing of the heavy and light chain, such as for example described in
  • the present invention provides novel engineered CH3 domains as well as novel combinations of CH3 mutations.
  • charged contact amino acids of CH3 domains that were known to be involved in CH3-CH3 pairing were substituted by amino acids of opposite charge (charge reversal), thereby influencing the CH3-CH3 pairing.
  • the mutations according to the present invention are an inventive alternative to this approach, because now CH3 amino acids that are non- charged or neutral in wildtype CH3 are substituted with charged residues.
  • the present invention in this embodiment does not exchange charged contact amino acids by amino acids of opposite charge but substitutes non-charged CH3 amino acids for charged ones.
  • the approach of the present invention provides not only a method for efficiently steering the dimerization of CH3 domains but also has the advantage that at least one additional charge-charge interaction in the CH3 interface is created.
  • the dimers according to the invention are generally more stable as compared to the wild type dimers (the wild type dimer is defined as a bispecific IgG (AB) without CH3 engineering in contrast to its parental homodimers (AA or BB)).
  • AB bispecific IgG
  • AA or BB parental homodimers
  • Example 17 discloses a method using mutations according to the present invention, wherein the proportion of a bispecific antibody of interest was raised to such extent that no dimeric by-product was detectable in the resulting mixture at all.
  • Unpaired half-molecules consisting of only a single heavy chain paired with a common light chain were present to some extent in the mixtures, but these are the result of unbalanced expression of the heavy chains and can be easily separated from the mixture by size exclusion chromatography.
  • a bispecific Ig-like molecule can be produced in a single cell with a high proportion with essentially no contaminating dimeric by-products being present, which is particularly suitable for the production of a pharmaceutical composition.
  • One preferred embodiment of the present invention therefore provides a method for producing a heterodimeric Ig-like molecule from a single cell, wherein said Ig-like molecule comprises two CH3 domains that are capable of forming an interface, said method comprising providing in said cell
  • said method further comprising the step of culturing said host cell and allowing for expression of said two nucleic acid molecules and harvesting said heterodimeric Ig-like molecule from the culture.
  • Said method preferably further comprises the step of providing said host cell with a nucleic acid molecule encoding a common light chain, which has advantages as outlined herein before.
  • the amino acids at position 366 of one CH3 domain and position 351 of a second CH3 domain have been reported to be a pair of contact residues in the CH3-CH3 interface, meaning that they are located sufficiently close to each other in the three-dimensional conformation of the resulting Ig-like molecule in order to be capable of interacting with each other.
  • the first CH3 domain will preferentially pair with the second CH3 domain.
  • threonine (T) at position 366 of a first CH3 domain is replaced by a first charged amino acid and leucine (L) at position 351 of a second CH3 domain is replaced by a second charged amino acid, wherein said first and second charged amino acids are of opposite charge.
  • first CH3 domain-comprising polypeptide, that carries a charged residue at position 366 further comprises a variable domain which has specificity for antigen A
  • second CH3 domain-comprising polypeptide, that carries an oppositely charged residue at position 351 further comprises a variable domain which has specificity for antigen B, bispecific Ig-like molecules with an AB specificity will be predominantly formed.
  • said means for preferential pairing of said 1 st and 2 nd CH3 domain-comprising polypeptides or said means for preferential pairing of said 3 rd and 4 th CH3 domain-comprising polypeptides are a substitution of threonine at position 366 of said 1 st or 3 rd CH3 domain by a first charged amino acid and substitution of leucine at position 351 of said 2 nd or 4 th CH3 domain by a second charged amino acid, wherein said first and second charged amino acids are of opposite charge.
  • One preferred combination of mutations according to the present invention is the substitution of threonine (T) by lysine (K) at position 366 of a first CH3 domain- comprising polypeptide which further comprises a variable domain (for instance with specificity A) and the substitution of leucine (L) by aspartic acid (D) at position 351 of a second CH3 domain-comprising polypeptide which further comprises a variable domain (for instance with specificity B).
  • T threonine
  • K lysine
  • L leucine
  • D aspartic acid
  • This is denoted as a T366K/L351'D pair mutation.
  • the amino acids at position 366 of one CH3 domain and position 351 of a second CH3 domain have been reported to be a pair of contact residues in the CH3-CH3 interface.
  • the lysine that is introduced at position 366 and the aspartic acid introduced at position 351 have opposite charges, so that these amino acids will electrostatically attract each other.
  • the first CH3 domain will preferentially attract the second CH3 domain and Ig-like molecules comprising a first CH3 domain containing lysine at position 366 paired with a second CH3 domain containing aspartic acid at position 351 will be predominantly formed. If the first CH3 domain-comprising polypeptide has specificity for antigen A, and if the second CH3 domain-comprising polypeptide has specificity for antigen B, bispecific Ig-like molecules with ⁇ ' specificity will be predominantly formed.
  • variable domains of both said first and second CH3-domain comprising polypeptide chains may be the same, which will result in the formation of monospecific Ig-like molecules (for instance with 'AA' specificity) .
  • one of the advantages of the mutations according to the present invention is the fact that a novel interaction between a newly introduced pair of charged amino acids is created, instead of replacing existing charged amino acid interactions. This was not previously disclosed or suggested.
  • One aspect of the invention therefore provides a method according to the present invention for producing at least two different Ig-like molecules from a single host cell, wherein said 1 st CH3 domain-comprising polypeptide chain comprises the amino acid substitution T366K, and said 2 nd CH3 domain-comprising polypeptide chain comprises the amino acid substitution L351D.
  • One embodiment provides a method for producing a heterodimeric Ig-like molecule from a single cell, wherein said Ig-like molecule comprises two CH3 domains that are capable of forming an interface, said method comprising providing in said cell:
  • said method further comprising the step of culturing said host cell and allowing for expression of said two nucleic acid molecules and harvesting said heterodimeric Ig-like molecule from the culture.
  • One embodiment therefore provides a method for producing a heterodimeric Ig-like molecule from a single cell, wherein said Ig-like molecule comprises two CH3 domains that are capable of forming an interface and wherein the presence of contaminating homodimers is less than 5%, preferably less than 2%, more preferably less than 1%, and most preferably contaminating homodimers are essentially absent, said method comprising providing in said cell:
  • said method further comprising the step of culturing said host cell and allowing for expression of said at two nucleic acid molecules and harvesting said heterodimeric Ig-like molecule from the culture.
  • a method according to the present invention for producing at least two different Ig-like molecules, or a method according to the invention for producing a heterodimeric Ig-like molecule wherein said first CH3-domain comprising polypeptide chain further comprises the amino acid substitution L351K. It is further preferred that said second CH3- domain comprising polypeptide chain further comprises an amino acid substitution selected from the group consisting of Y349E, Y349D and L368E. Most preferably said second CH3-domain comprising polypeptide chain further comprises the amino acid substitution L368E.
  • T366K/L351'D mutations according to the present invention are further combined with the substitution of leucine (L) by glutamic acid (E) at position 368 of the second CH3 domain.
  • L leucine
  • E glutamic acid
  • T366K/L351'D,L368'E mutation but alternative ways of denoting are also possible, such as T336K/L351D-L368E or T366K/L351D,L368E or T366K - L351D,L368E).
  • a particularly preferred embodiment therefore provides a method for producing a heterodimeric Ig-like molecule from a single cell, wherein said Ig-like molecule comprises two CH3 domains that are capable of forming an interface and wherein the presence of contaminating homodimers is less than 5%, preferably less than 2%, more preferably less than 1%, and most preferably contaminating homodimers are essentially absent, said method comprising providing in said cell: - a first nucleic acid molecule encoding a 1 st CH3 domain-comprising polypeptide chain, and
  • said method further comprising the step of culturing said host cell and allowing for expression of said at two nucleic acid molecules and harvesting said heterodimeric Ig-like molecule from the culture.
  • threonine (T) is substituted by lysine (K) at position 366 of a first CH3 domain and leucine (L) is substituted by aspartic acid (D) at position 351 of a second CH3 domain and tyrosine (Y) is substituted by glutamic acid (E) at position 349 of said second CH3 domain.
  • T366K/L351'D,Y349'E mutation but other ways of denoting these mutations may include for example T366K - L351D:Y349E, or T366K/L351D,Y349E or simply T366K/L351DY349E.
  • Residue Y349 is a neighboring residue of the residue at position 351 that may contribute to dimer interactions. According to in silico data, Y349E adds to the stability of the heterodimer (lower in silico scores) as well as to the destabilization of the monodimer (higher in silico scores) and glutamic acid (E) on position 349 is more favorable than aspartic acid (D).
  • introduction of a second amino acid substitution in the second CH3 domain comprising polypeptide, comprising already the amino acid substitution at position 351 favors
  • a particularly preferred embodiment therefore provides a method for producing a heterodimeric Ig-like molecule from a single cell, wherein said Ig-like molecule comprises two CH3 domains that are capable of forming an interface and wherein contaminating homodimers are less than 5%, more preferably less than 2%, even more preferably less than 1%, and most preferably essentially absent, said method comprising providing in said cell:
  • said method further comprising the step of culturing said host cell and allowing for expression of said at two nucleic acid molecules and harvesting said heterodimeric Ig-like molecule from the culture.
  • threonine (T) is substituted by lysine (K) at position 366 of a first CH3 domain and leucine (L) is substituted by aspartic acid (D) at position 351 of a second CH3 domain and tyrosine (Y) is substituted by glutamic acid (E) at position 349 of said second CH3 domain and leucine (L) is substituted by glutamic acid (E) at position 368 of said second CH3 domain.
  • T366K/L351'D,Y349'E,L368'E mutation is residues that may contribute to dimer interactions.
  • Y349E and L368E add to the stability of the heterodimer (lower in silico scores) as well as to the destabilization of the BB dimer (higher in silico scores) and glutamic acids (E) on positions 349 and 368 are more favorable than aspartic acids (D).
  • a particularly preferred embodiment therefore provides a method for producing a heterodimeric Ig-like molecule from a single cell, wherein said Ig-like molecule comprises two CH3 domains that are capable of forming an interface and wherein contaminating homodimers are less than 5%, more preferably less than 2%, even more preferably less than 1%, and most preferably essentially absent, said method comprising providing in said cell:
  • said method further comprising the step of culturing said host cell and allowing for expression of said at two nucleic acid molecules and harvesting said heterodimeric Ig- like molecule from the culture.
  • threonine (T) is substituted by lysine (K) at position 366 of a first CH3 domain and leucine (L) is substituted by lysine (K) at position 351 of said first CH3 domain and leucine (L) is substituted by aspartic acid (D) at position 351 of a second CH3 domain and leucine (L) is substituted by glutamic acid (E) at position 368 of said second CH3 domain.
  • T366K,L351K/L351'D,L368'E mutation This mutation also enhances the proportion of the (bispecific) antibody of interest, as shown in the Examples. Also with this mutation it has become possible to obtain bispecific antibody without any detectable amount of homodimers formed.
  • a method for producing a heterodimeric Ig-like molecule from a single cell wherein said Ig-like molecule comprises two CH3 domains that are capable of forming an interface and wherein contaminating homodimers are less than 5%, preferably less than 2%, more preferably less than 1%, and most preferably essentially absent, said method comprising providing in said cell:
  • said method further comprising the step of culturing said host cell and allowing for expression of said at two nucleic acid molecules and harvesting said heterodimeric Ig-like molecule from the culture.
  • threonine (T) is substituted by lysine (K) at position 366 of a first CH3 domain and leucine (L) is substituted by lysine (K) at position 351 of said first CH3 domain and leucine (L) is substituted by aspartic acid (D) at position 351 of a second CH3 domain and tyrosine (Y) is substituted by aspartic acid (D) at position 349 of said second CH3 domain and arginine (R) is substituted by aspartic acid (D) at position 355 of said second CH3 domain.
  • T366K,L351K/L351'D,Y349'D,R355'D mutation is denoted as a T366K,L351K/L351'D,Y349'D,R355'D mutation.
  • the T366K-L351K/L351'D-Y349'D pair may be further improved by the R355'D mutation in the B-chain, which results in a higher BB-in silico score, but also the AB in silico score is slightly higher.
  • a method for producing a heterodimeric Ig-like molecule from a single cell wherein said Ig-like molecule comprises two CH3 domains that are capable of forming an interface and wherein contaminating homodimers are less than 5%, more preferably less than 2%, even more preferably less than 1%, and most preferably essentially absent, said method comprising providing in said cell:
  • said method further comprising the step of culturing said host cell and allowing for expression of said at two nucleic acid molecules and harvesting said heterodimeric Ig-like molecule from the culture.
  • Table B provides an overview of mutations that can be introduced in CH3 domains as preferred means for preferential pairing to create either heterodimers or homodimers.
  • said means for preferential pairing of said 1 st and 2 nd CH3 domain-comprising polypeptides and said means for preferential pairing of said 3 rd and 4 th CH3 domain-comprising polypeptides comprise at least two combinations of mutations as depicted in Table B.
  • the present invention also provides novel combinations of CH3 mutations with which it has become possible to produce a mixture of at least two monospecific Ig-like molecules in a single cell, wherein contaminating bispecific Ig-like molecules are less than 5%, preferably more than 2%, even more preferably less than 1%, and most preferably even essentially absent.
  • mutations according to the invention are, therefore, particularly suitable for the production of a mixture of monospecific antibodies, which is for instance advantageous when a high level of crosslinking of two identical target molecules is desired, when the density of antibodies on a target cells needs to be high enough to recruit certain effector functions such as complement- mediated lysis of a tumor cell, or when two targets are located too far away from each order so that they cannot be bound by as single bispecific antibody, or in order to simplify regulatory approval procedures. In such cases, it is often desired to optimize the production platform for such monospecific antibodies.
  • the present invention provides the insight that when lysine (K) at position 392 of a first CH3 domain-comprising polypeptide (for instance having specificity A) is substituted by aspartic acid (D) and when aspartic acid (D) at position 399 of said first CH3 domain-comprising polypeptide is substituted by lysine (K) and when lysine (K) at position 409 of said first CH3 domain-comprising polypeptide is substituted by aspartic acid (D), it has become possible to produce a mixture of at least two different monospecific Ig-like molecules in a single cell, including monospecific Ig-like molecules with specificity AA, wherein the formation of bispecific by-products
  • substitutions, or other functional variants such as e.g. K392D and K409D, D399R and K409E and so forth, may also result in similar effects.
  • E356K, E357K, K439D, K370D is also particularly preferred for the production of a mixture of monospecific Ig-like molecules.
  • functional variants thereof i.e., E356R, E357R, K439E, K370E
  • triple or double mutants comprising E356K and K439D, and E357K and K370D substitutions, or other functional variants may also result in similar effects.
  • a further embodiment therefore provides a method for producing at least two different monospecific Ig-like molecules from a single host cell, wherein each of said two Ig-like molecules comprises two CH3 domains that are capable of forming an interface, said method comprising providing in said cell
  • said method further comprising the step of culturing said host cell and allowing for expression of said nucleic acid molecules and harvesting said at least two different Ig-like molecules from the culture.
  • An alternative embodiment provides a method for producing at least two different monospecific Ig-like molecules from a single host cell, wherein each of said two Ig-like molecules comprises two CH3 domains that are capable of forming an interface, said method comprising providing in said cell
  • said first CH3 domain-comprising polypeptide chain comprises either a wildtype CH3 domain or comprises a K392D, D399K, K409D mutation and said second CH3 domain-comprising polypeptide chain comprises a E356K, E357K,
  • said method further comprising the step of culturing said host cell and allowing for expression of said nucleic acid molecules and harvesting said at least two different Ig-like molecules from the culture.
  • two monospecific Ig-like molecules can be produced in a single cell, wherein the formation of bispecific Ig-like molecules is essentially undetectable.
  • the skilled person may select a 3 rd nucleic acid molecule encoding a wildtype or engineered CH3 domain-comprising polypeptide chain to provide to said host cell such that a mixture of 3 monospecific antibodies is produced, and so forth.
  • a method according to the invention for producing at least two different Ig-like molecules or for producing a heterodimeric Ig-like molecule wherein each of the CH3-domain comprising polypeptide chains further comprises a variable region recognizing a different target epitope, wherein the target epitopes are located on the same molecule. This often allows for more efficient counteraction of the (biological) function of said target molecule as compared to a situation wherein only one epitope is targeted.
  • a heterodimeric Ig-like molecule may simultaneously bind to 2 epitopes present on, e.g., growth factor receptors or soluble molecules critical for tumors cells to proliferate, thereby effectively blocking several independent signalling pathways leading to uncontrolled proliferation, and any combination of at least two Ig-like molecules may
  • the target molecule is a soluble molecule. In another preferred embodiment, the target molecule is a membrane-bound molecule.
  • each of the CH3-domain comprising polypeptide chains further comprises a variable region recognizing a target epitope, wherein the target epitopes are located on different molecules.
  • each of the different target molecules may either be a soluble molecule or a membrane-bound molecule.
  • the different target molecules are soluble molecules.
  • one target molecule is a soluble molecule whereas the second target molecule is a membrane bound molecule.
  • both target molecules are membrane bound molecules.
  • the different target molecules are expressed on the same cells, whereas in other embodiments the different target molecules are expressed on different cells.
  • any combination target molecules are expressed on different cells.
  • heterodimeric Ig-like molecule or any combination of at least two Ig-like molecules may be suitable for simultaneously blocking multiple membrane-bound receptors, neutralizing multiple soluble molecules such as cytokines or growth factors for tumor cells or for neutralizing different viral serotypes or viral strains.
  • One preferred embodiment provides a method according to the invention for producing at least two different Ig-like molecules or for producing a heterodimeric Ig- like molecule, wherein at least one of said target epitopes is located on a tumor cell. Alternatively, or additionally, at least one of said target epitopes is located on the surface of an effector cell. This is for instance suitable for recruitment of T cells or NK cells for tumor cell killing.
  • at least one Ig-like molecule is produced with a method according to the invention that is capable of recruiting immune effector cells, preferably human immune effector cells, by specifically binding to a target molecule located on immune effector cells.
  • said immune effector cell is activated upon binding of the Ig-like molecule to the target molecule.
  • effector mechanisms may for instance encompass the redirection of immune modulated cytotoxicity by administering an Ig-like molecule produced by a method according to the invention that is capable of binding to a cytotoxic trigger molecule such as the T cell receptor or an Fc gamma receptor, thereby activating downstream immune effector pathways.
  • a cytotoxic trigger molecule such as the T cell receptor or an Fc gamma receptor
  • the term 'immune effector cell' or 'effector cell' as used herein refers to a cell within the natural repertoire of cells in the mammalian immune system which can be activated to affect the viability of a target cell.
  • Immune effector cells include cells of the lymphoid lineage such as natural killer (NK) cells, T cells including cytotoxic T cells, or B cells, but also cells of the myeloid lineage can be regarded as immune effector cells, such as monocytes or macrophages, dendritic cells and neutrophilic granulocytes.
  • said effector cell is preferably an NK cell, a T cell, a B cell, a monocyte, a macrophage, a dendritic cell or a neutrophilic granulocyte.
  • Target antigens present on immune effector cells may include CD 3, CD 16, CD25, CD28, CD64, CD89, NKG2D and NKp46. Further provided is therefore a method according to the invention for producing at least two different Ig-like molecules or for producing a heterodimeric Ig-like molecule, wherein said target epitope is located on a CD 3, CD 16, CD25, CD28, CD64, CD89, NKG2D or a NKp46 molecule.
  • the viability of a target cell may include cell survival, proliferation and/or ability to interact with other cells.
  • the present invention thus provides methods according to the invention for producing a heterodimeric Ig-like molecule, wherein each of the CH3-domain comprising polypeptide chains further comprises a variable region recognizing a target epitope.
  • each of the 2 variable regions of the CH3-domain comprising polypeptide chains recognizes the same target epitope but with different affinities.
  • each of the 2 variable regions of the CH3-domain comprising polypeptide chains recognizes a different target epitope.
  • the different target epitopes are located on the same target molecule, which can be either a membrane-bound molecule or a soluble molecule.
  • the different target epitopes are located on different target molecules, which can be either expressed on the same cells or on different cells.
  • the different target molecules can be soluble molecules, or one target molecule can be a soluble molecule whereas the second target molecule is a membrane bound molecule.
  • at least one of the target molecules of the heterodimeric Ig-like molecule is located on a tumor cell.
  • at least one of the target molecules of the heterodimeric Ig-like molecule is located on an effector cell (i.e.
  • an NK cell, a T cell, a B cell, a monocyte, a macrophage, a dendritic cell or a neutrophilic granulocyte, and said target epitope may be located on a CD3, CD 16, CD25, CD28, CD64, CD89, NKG2D or a NKp46 molecule).
  • a method according to the invention for producing at least two different Ig-like molecules or for producing a heterodimeric Ig-like molecule is provided, wherein said at least two different Ig-like molecules are antibodies, most preferably antibodies of the IgG isotype, even more preferably the IgGl isotype, as described herein above.
  • an Ig-like molecule, a heterodimeric Ig-like molecule, or a mixture of at least two Ig-like molecules obtainable by a method according to the present invention.
  • Said (heterodimeric) Ig-like molecule or mixture of Ig-like molecules preferably comprises at least one CH3 mutation as depicted in Table B.
  • Ig-like molecule or a mixture of at least two Ig-like molecules, comprising at least one mutation as depicted in Table B is therefore also herewith provided, as well as a pharmaceutical composition comprising at least one Ig-like molecule, or a mixture of at least two Ig-like molecules, according to the present invention.
  • said Ig-like molecule is a bispecific Ig-like molecule, such as a bispecific antibody.
  • said Ig-like molecule is a monospecific Ig-like molecule, such as a monospecific antibody.
  • One preferred embodiment provides a mixture of at least two different Ig-like molecules obtainable by a method according to the invention, wherein said at least two different Ig-like molecules bind to different epitopes on the same antigen and/or to different epitopes on different antigens.
  • a heterodimeric Ig-like molecule obtainable by a method according to the invention, wherein said heterodimeric Ig-like molecule binds to different epitopes on the same antigen and/or to different epitopes on different antigens.
  • the invention also provides a mixture of at least two different Ig-like molecules obtainable by a method according to the invention, wherein said at least two different Ig-like molecules comprise at least one heterodimeric Ig- like molecule.
  • two of said at least two different Ig-like molecules are heterodimeric Ig-like molecules.
  • Yet another preferred embodiment provides a heterodimeric antibody comprising two CH3 domains, wherein one of said two CH3 domains comprises the amino acid substitutions L351D and L368E and wherein the other of said two CH3 domains comprises the amino acid substitutions T366K and L351K. These amino acid substitutions are preferred means for preferential pairing of said two CH3 domains, as explained before.
  • the amino acid substitutions L351D and L368E in one of said two CH3 domains and the amino acid substitutions T366K and L351K in the other of said two CH3 domains are together dubbed the 'DEKK combination of mutations', 'DEKK variant', 'DEKK pair', 'DEKK engineered CH3 domains', 'DEKK' or alternative names referring to DEKK are used.
  • the CH3 domain that carries the amino acid substitutions L351D and L368E is also dubbed 'the DE- side' and the CH3 domain that carries the amino acid substitutions T366K and L351K is also dubbed 'the KK-side'.
  • a pharmaceutical composition comprising a (heterodimeric) Ig-like molecule, or a mixture of at least two Ig-like molecules obtainable by any method according to the invention.
  • Said (heterodimeric) Ig-like molecule, or said at least two Ig-like molecules according to the invention is/are preferably (an) antibody/antibodies.
  • Said pharmaceutical composition may comprise said (heterodimeric) Ig-like molecule, a mixture comprising monospecific or bispecific Ig-like molecules, or a combination of monospecific and bispecific Ig-like molecules.
  • a pharmaceutical composition according to the invention comprises a pharmaceutically acceptable carrier.
  • such 'pharmaceutically acceptable carrier includes any and all solvents, salts, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible.
  • route of administration e.g., intravenously,
  • the Ig-like molecules may be coated in a material to protect the Ig-like molecules from the action of acids and other natural conditions that may inactivate the Ig-like molecules.
  • a pharmaceutical composition comprising a mixture of at least two Ig-like molecules obtainable by any method according to the invention is provided, wherein said at least two different Ig- like molecules have been produced by recombinant host cells according to the present invention.
  • a pharmaceutical composition comprising a heterodimeric Ig-like molecule obtainable by any method according to the invention, wherein said heterodimeric Ig-like molecule has been produced by recombinant host cells according to the present invention.
  • a nucleic acid molecule encoding a CH3 domain-comprising polypeptide chain that comprises at least one mutation as depicted in Table B is also provided herewith, as well as a recombinant host cell comprising at least one nucleic acid molecule encoding a CH3 domain-comprising polypeptide chain that comprises at least one mutation as depicted in Table B.
  • Figure 1 A) schematic representation of construct vector MV1057.
  • the stuffer region is the region into which an antibody VH region is cloned.
  • Figure 2 amino acid sequence of wildtype IgGl Fc, as present in construct vector MV1057 (EU numbering scheme applied).
  • Figure 3 nucleotide and amino acid sequences of VH regions used for cloning into the various constructs.
  • Figure 4 mass spec data of transfections A, G and H.
  • Figure 7 prevention of homodimerisation by substitution of neutral amino acids for charged amino acids.
  • FIG. 8 Native MS spectrum of transfection sample ZO (T366K/L351'D) (A) and
  • Figure 10 Cartoons of interactions in the CH3-CH3 interface
  • K409D:K392D/D399'K:E356'K B) D399K:E356K/D399'K:E356'K
  • Figure 12 Cartoons of interactions in the CH3-CH3 interface ; A) L351D/L351'D, B) L351D:S354A:R355D/ L351'D:S354'A:R355'D
  • Figure 14 HADDOCK scores for additional charge mutations around position T366 in chain A and position L351 in chain B.
  • FIG. 15 Cartoons of interactions in the CH3-CH3 interface
  • Figure 18 Examples of nMS spectra for bispecific IgG obtained after the co-expression of construct T366K,L351K with either construct L351D (left hand panel) or L351D,Y349E (right hand panel), zoomed in on a single charge state of the full IgG (half bodies not shown)
  • Figure 19 A) Results of native MS showing relative abundances of AA, AB, BB, A and B (total of all species is 100%); B) idem but now without AB to have a better overview on the undesired species AA, BB, A and B
  • Figure 20 Results of thermostability assay. Squares: wildtype; triangles: charge reversal pair E356K:D399K/K392D:K409D; circles: mutant CH3 combinations as indicated above each graph.
  • Figure 22 Results in serum stability, measured by ELISA using fibrinogen as coated antigen.
  • Figure 23 nMS results of ratio experiments with transfection ratios from 1:5 to 5: 1.
  • Figure 25 HADDOCK scores for dimers with different CH3 engineered vectors. Grey bars: Desired species AB and CD; black bars: undesired species AA, BB, CC, DD, AC,
  • Figure 26 SDS-PAGE of transfections # 1-11 from Table 20. Control samples DE/KK, DE/DE and KK/KK are also included.
  • Figure 27 nMS of transfections # 9 (A) and # 11 (B).
  • Figure 28 nMS of gel filtrated samples 1516: 1516 (A), 1337: 1337 (B) and 1516: 1337 (C).
  • Figure 29 serum levels of samples of DEKK engineered antibody and its two parental antibodies (pK study).
  • Example 1 amino acid substitutions to create various different CH3- domains
  • construct vector MV1057 comprises nucleic acid sequences encoding the normal wildtype IgGl Fc part, as depicted in figure 2.
  • Table 1 lists the amino acid substitutions that were introduced in this wildtype Fc, resulting in a series of seven constructs. All constructs were made at Geneart. Constructs 1, 2 and 3, or alternatives thereof, have previously been described to drive
  • CH3 variants can be used in association with other antibody domains to generate full length antibodies that are either bispecific or monospecific.
  • the specificity of the antibody as defined by the VH/VL combinations will not affect the heavy chain dimerization behaviour that is driven by the CH3 domains.
  • Model VH/VL combinations were used throughout the studies, wherein all VLs are based on the germline human IGKV1-39 and VHs vary.
  • Figure 3 provides full sequences and specificities of the antibody VH regions used throughout the studies.
  • the MF coding refers to internal Merus designation for various VHs, e.g.
  • VH MF1337 has specificity for tetanus toxoid, MF1025 for porcine thyro globulin, MF1122 for bovine fibrinogen.
  • VH regions present in phage display vector MV1043 are digested with restriction enzymes Sfil and BstEII (New England Biolabs/ cat# R0123L and R0162L/ according to manufacturer's instructions) that release the VH fragment from this vector.
  • Vector MV1057 is digested with Sfil and BstEII according to standard procedures (according to manufacturer's instructions).
  • Fragments and vector are purified over gel (Promega/ cat# V3125/ according to manufacturer's instructions) to isolate the cut vector and VH gene inserts. Both are combined by ligation after which the ligation is transformed into E. coli DH5a (Invitrogen/ cat# 12297-016/ according to manufacturer's instructions). After overnight selection single colonies are picked and vectors with a correct insert identified by sequencing.
  • Example 3 transfection and expression of full IgG in HEK293T cells
  • Culture supernatants were purified using protein A columns (GE Healthcare/ cat# 11- 0034-95/ according to manufacturer's instructions) and eluted in 0, 1 M citrate buffer pH 3.0 and immediately neutralized in an equal volume of 1,0 M Tris-HCL pH 8.0 or directly rebuffered to PBS using a desalting column.
  • protein A beads sepharose beads CL-4B, GE healthcare cat #170780-01
  • Antigen specific ELISAs were performed to establish binding activity against the antigens and capture ELISAs were carried out to demonstrate binding activity of the bispecific antibodies. Biotinylated second antigen was used for detection of the complex, (de Kruif etal Biotech Bioeng. 2010)
  • Example 6 SDS-PAGE
  • the purified IgG mixtures were analysed by SDS-PAGE (NuPAGE® 4- 12% bis-tris gel/ Invitrogen/ cat# NP0323BOX) under reduced and non-reducing conditions according to standard procedures, and staining of proteins in gel was carried out with colloidal blue (PageBlueTM protein staining solution/ Fermentas/ cat# R0571).
  • N-glycosidase F N-glycosidase F
  • Example 8 Native mass spectrometric analysis
  • Mass Spectrometry was used to identify the different IgG species in the purified IgG mixtures and to establish in what ratios these IgG species are present. Briefly, 2-3 ⁇ at a 1 ⁇ concentration in 150 mM ammonium acetate pH 7.5 of IgG's were loaded into gold-plated borosilicate capillaries made in-house (using a Sutter P-97 puller [Sutter Instruments Co., Novato, CA, USA] and an Edwards Scancoat six sputter- coater [Edwards Laboratories, Milpitas, CA, USA]) for analysis on a LCT 1 mass spectrometer (Waters Corp., Milford, MA, USA), adjusted for optimal performance in high mass detection (Tahallah et al., RCM 2001).
  • a capillary voltage of 1300 V was used and a sampling cone voltage of 200 V; however, these settings were adjusted when a higher resolution of the 'signal-to-noise' ratio was required.
  • the source backing pressure was elevated in order to promote collisional cooling to approximately 7.5 mbar.
  • MassLynx 4.1 Waters Corp., Milford, MA, USA. Minimal smoothing was used, after which the spectra were centered. The mass of the species was calculated using each charge state in a series. The corresponding intensities of each charge state were assigned by MassLynx and summed. This approach allowed the relative quantification of all species in a sample. Alternatively, quantification of the peaks can be performed using area-under-the-curve (AUC) methods, known in the art. All analyses were repeated three times to calculate standard deviations of both the masses of the IgG's as well as their relative abundance.
  • AUC area-under-the-curve
  • Example 10 mixtures of 2 or 3 monospecific antibodies from a single cell Several antibody VH regions with known specificities and known ability to pair with the human IGKV1-39 light chain ( Figure 3) were used for recloning into the wildtype construct vector MV1057, or in construct 4 or construct 5 of Table 1, resulting in vectors I-III (Table 2).
  • Table 3 depicts the transfection schedule and results.
  • transfections A, G and H resulted in formation of homodimers only, and 100% of bivalent monospecific AA, BB or CC was retrieved from cells transfected with any one of vectors I, II or III (Fig. 4).
  • this was to be expected and previously demonstrated for transfection A it is actually now shown for the first time that homodimerisation of CH3-engineered Ig heavy chains containing either the triple amino acid substitution of construct 4 (i.e., K392D, D399K, K409D) or the quadruple amino acid substitution of construct 5 (i.e., E356K, E357K, K439D, K370D) is reported (transfections G and H).
  • co-expression experiments of two vectors in a single cell were performed.
  • transfections M and N show that wildtype and CH3 engineered Ig heavy chains can be co-expressed in a single cell together with a common light chain resulting in mixtures of two species of monospecific antibodies without the presence of undesired bispecific antibodies and with as little as 4-5% contaminating 'other molecules' present in the mixture.
  • Other molecules' is defined as all molecules that do not have the mass of an intact IgG, and includes half molecules consisting of a single heavy and light chain pair.
  • the fraction 'other' does not include bispecific product.
  • the ratio of AA:BB was close to 1: 1 upon transfection of equal ratios of vector DNA.
  • transfection N resulted in an almost 10: 1 ratio of AA:CC.
  • transfection U was repeated with adjusted ratios of DNA
  • transfection U a 1:5 ratio of vector DNA I:III equalized the ratio of AA:CC antibody product in the mixture towards an almost 1: 1 ratio.
  • transfections M and U show that it is possible to express two different, essentially pure, monospecific antibodies in a single cell, without undesired by products (i.e., no abundant presence of AC or half molecules A or C) (fig. 5).
  • the novel CH3 modifications of constructs 4 and 5 differ substantially from wildtype CH3 such that heterodimerization between wildtype and 4, or wildtype and 5, does not occur, which is advantageous for application in large scale production of mixtures of monospecific antibodies from single cells.
  • Transfections with altered vector DNA ratios (1:1: 10, transfection V) demonstrated that ratios of AA:BB:CC in the mixtures can be steered towards desired ratios.
  • monospecific antibodies can be expressed in a single cell without undesired by products, offering advantages for large scale production of mixtures of therapeutic monospecific antibodies.
  • Example 11 mixtures of 2 bispecific antibodies from a single cell
  • Antibody VH regions with known specificities and known ability to pair with the human IGKV1-39 light chain were used for recloning into vectors containing constructs 1-3 or 6-7 of Table 1 resulting in vectors IV-X (Table 4).
  • Vectors IV-X each containing nucleic acid sequences encoding the common human light chain as well as an Ig heavy chain with different CH3 region and different VH specificity, were subsequently transfected into cells, either alone to demonstrate that formation of intact monospecific antibodies was hampered, or in combination with another construct vector to obtain bispecific antibodies or mixtures of two bispecific antibodies.
  • Table 5 depicts the transfection schedule and results.
  • CH3-engineered Ig heavy chains encoded by constructs 1 and 2 are still able to form homodimers when expressed alone in single cells (WO2009/089004).
  • WO2009/089004 further reports that CH3 domains that are engineered to comprise triple charge pair mutations, such as present in construct 3, are no longer capable of forming homodimers when expressed alone.
  • a fourth antibody VH region with known specificity and known ability to pair with the human IGKV1- 39 light chain will be used for recloning into vectors containing constructs 1-3 or 7 of Table 1, resulting in vectors ⁇ , ⁇ , ⁇ or X' (the ' indicating a different specificity as compared to corresponding vector numbers).
  • the resulting vectors ⁇ - ⁇ , X' and IV- IX each containing nucleic acid sequences encoding for the common human light chain as well as an Ig heavy chain with different CH3 region and different VH specificity, will subsequently be transfected into cells, in combination with other construct vectors to obtain a variety of mixtures of bispecific and/or monospecific antibodies.
  • the variety of mixtures that will be obtained include mixtures of 2 bispecific antibodies recognizing 4 epitopes, 2 bispecific antibodies and one monospecific antibody, or mixtures of 1 bispecific and one monospecific antibody from a single cell. Table 6 depicts the transfection schedule and expected results.
  • the objective of this study was to engineer the IgG CH3 region to result in the production of only heterodimers or only homodimers upon mixed expression of different IgG heavy chains in a single cell, wherein the novel engineered CH3 domains will not homo- or heterodimerize with known engineered CH3 domains, or with wildtype CH3 domains. Therefore, as a first step in identifying novel engineered CH3 domains that would meet the criteria, many interface contact residues in the IgG CH3 domain were scanned one by one or in groups for substitutions that would result in repulsion of identical heavy chains - i.e., reduced homodimer formation - via electrostatic interactions.
  • the objective was to obtain a list of residues that, when substituted by a charged residue, would result in repulsion of identical chains such that these mutations may be used to drive homo- and/or heterodimer formation upon mixed expression of different IgG heavy chains, whereby the obtained full length IgGs are stable and are produced with high proportions.
  • the identified substitutions will be used to generate bispecific antibodies or mixtures of bispecific or monospecific antibodies by engineering matched pairs of CH3 residues in one or more IgG heavy chains - CH3 regions.
  • residues to be tested in the present study are contact residues as previously identified (Deisenhofer J., 1981; Miller S., 1990; Padlan, 1996, Gunasekaran, 2010).
  • the rationale for this approach is that repulsive charges are engineered into each available pair of contacting residues.
  • Amino acid substitutions were introduced in construct vector MV1057 by Geneart according to the table 7 and expression of constructs was performed by transfection in HEK293T cells, according to standard procedures. IgG expression levels were measured in Octet. When production failed twice, the mutation was considered to be detrimental to expression and the mutation was not pursued further.
  • transfection ZO resulted in fair amounts of bispecific product
  • the reverse charge pair of transfection ZP did not result in similar results, and only 52% of bispecific product was observed, with considerable amounts of the two homodimers being present (30% AA and 13% CC).
  • An explanation for this may be that the negatively charged D structurally closely resembles T, hence the T366D may not be potent enough to repulse itself and T366D will thus still form homodimers, as was indeed observed.
  • Example 14 HADDOCK for design of new CH3 mutants to drive efficient heterodimerization.
  • the newly found charge pair T366K/L351'D increases the proportion of heterodimers in the mixture (69 %) with a small fraction of undesired CC homodimers (7%) (L351D/L351'D) and a substantial fraction of half A molecules (24%) 'contaminating' the mixture.
  • an in silico approach was used to generate further insight in amino acid residues involved CH3 interface interactions, to test complementary substitutions in opposing CH3 regions and to find novel CH3 pairs containing complementary substitutions that further increase efficient heterodimerization while preventing efficient formation of homodimers of the two heavy chains.
  • HADDOCK High Ambiguity Driven protein-protein DOCKing
  • HADDOCK distinguishes itself from ab-initio docking methods in the fact that it encodes information from identified or predicted protein interfaces in ambiguous interaction restraints (AIRs) to drive the docking process, (de Vries et al., 2010).
  • the input for the HADDOCK web server consists of a protein structure file, which can be a crystal structure, NMR structure cluster or a modeled structure.
  • HADDOCK After the docking or refinement, HADDOCK returns a so-called HADDOCK score, which is a weighted average of VanderWaals energy, electrostatic energy, buried surface area 5 and desolvation energy.
  • the HADDOCK score can be interpreted as an indication of binding energy or affinity, even though a direct translation to experimental data is often hard to achieve.
  • HADDOCK provides structure files for the 'top four' structures that resulted from the docking run. These structure files can be downloaded and visualized, enabling the detailed analysis of the interactions of the 10 individual residues.
  • the HADDOCK output consists of a set of calculated energies, a HADDOCK score
  • the HADDOCK-scores are used to compare different structures; the other energies are merely used to get an indication about what is happening in the structures (e.g. good electrostatic
  • HADDOCK score the better. For each mutation pair, the scores were calculated for the AA, AB and BB dimers.
  • HADDOCK scores are the same for AA, AB and
  • the higher HADDOCK scores for the homodimers can thus be explained by the electrostatic repulsion of the mutated interface residues. These residues have to bend away from each other and don't have interaction with residues on the other chain, causing a drop in the affinity.
  • Table 11 and Figure 9 confirm what was observed in example 13.
  • the T366K/L351'D AC heterodimer and the L351D/L351'D CC homodimer form with a similar energy, explaining the presence of both the heterodimer and homodimer in the mixture.
  • the T366K/T366'K AA homodimer is barely detectable in the mixture although T366K half A molecules are present.
  • Table 11 and Figure 9 indeed show that the HADDOCK score for the T366K/T366'K AA homodimer is higher than the score for the AC heterodimer; hence formation of this homodimer is energetically less favorable.
  • Example 15 366/351 variations
  • T366K/L351'D mutant charge pair can be designed that may have similar results in terms of percentage of bispecific antibodies in the mixture.
  • Alternatives may include substitutions T366R, T366D, T366E, L351E, L351K and L351R.
  • the proportion of CC homodimers of L351D/L351'D may be diminished by creating variants of the 366/351 pair. All possible mutation pairs were run in HADDOCK and the resulting scores are presented in Table 12 and visualized in Figure 11.
  • T366K/L351'E pair were taken as a starting structure.
  • additional mutations on the B-chain were used to calculate the HADDOCK-scores and energies.
  • the T366K/L351'D pair was again taken as starting structure.
  • additional mutations were added to the A-chain which already comprises the T366K substitution. As shown in Figure 14, there are several mutation pairs that seem favorable towards the formation of bispecific heterodimers.
  • T366K-L351K/L351'D-Y349'D pair may be further improved by the R355'D mutation in the B-chain, which results in a higher BB-HADDOCK score, but also the AB HADDOCK score is slightly higher.
  • Overall the additional L351K results in lower AB scores and similar AA and BB scores when compared to the sole T366K mutation in the A chain. Theoretically this would result in higher amounts of bispecific heterodimers in the samples.
  • example 16 suggested that some CH3 variants with additional mutations around the T366K/L351'D pair would yield mixtures with higher proportions of the bispecific component and potentially lower proportions of the homodimeric component. These best performing pairs were selected for production and further analysis.
  • constructs T366R and L351E were also generated. Table 14 lists the constructs that were made and which were used for recloning antibody VH regions with known specificities and known ability to pair with the human IGKV1-39 light chain. Expression of the IgGs that contain the individual constructs was previously reported in example 13, and was repeated for the constructs as listed in Table 14. Aim was to assess which of the constructs homodimerize in the absence of a matching heterodimerization partner.
  • E356K:D399K/K392'D:K409'D and E356K:D399K/K392'D:K409'D:K439'D charge reversal pairs, in that the previously described charge variants are based on the reversal of existing charges within the CH3-CH3 interface whereas the newly identified charge variants are adding additional charge pairs (charge -charge interactions) to the CH3-CH3 interface.
  • the introduction of additional charge pairs in the CH3-CH3 interface may further increase the stability of the interface and thereby of the intact antibody.
  • Intrinsic fluorescence spectra will be monitored at 25°C using a FluoroMax
  • the fluorescence method will be optimized.
  • the fluorescence emission will provide information on conformation and aggregation properties.
  • Excitation and emission slits will be optimized. For example, right angle light-scattering can distinguish between IgG samples that have no and 5% dimers.
  • Nile Red in ethanol will be added to the sample.
  • the samples will be filled in a microscopy slide and analyzed by fluorescence microscopy. Particles will be counted.
  • the lower size limit of the particles that can be observed by fluorescence microscopy is approximately 0.5 ⁇ .
  • Thermo-stability studies using the Octet are explored, both with Protein A biosensors and by using FcRn binding to IgG.
  • the samples will be incubated at a concentration of 100 ug/ml (in PBS) at 4, 50, 55, 60, 65, 70 and 75°C for 1 hour using a PCR machine. Following this the samples will be cooled down slowly during a period of 15 minutes to 25°C and kept at this temperature for 2 hours, after which they will be stored overnight at 4°C.
  • Precipitated antibodies will be removed by centrifugation, after which the total IgG concentration of soluble antibodies will be determined by Octet using the protein A Biosensor (1/10 dilution in PBS). Assays that measure binding of the CH3 engineered IgG to FcRn using the Octet are being explored. Either protein L biosensors are used to bind the light chain of IgG to the sensor, followed by incubation with FcRn in solution, or anti-penta-HIS biosensors are used to bind His-tagged FcRn protein, followed by incubation with the IgG of interest. These methods may be more sensitive than using the protein A Biosensor and can also be used for thermal stability studies. All samples will also be analyzed for serum stability.
  • IgG samples will be incubated at 37°C in human serum, control samples will be kept at 4°C. After 1, 2, 3 and 4 weeks, samples are centrifuged to remove precipitated IgG. Subsequently the sample is titrated in antigen-specific ELISA to determine the relative amounts of functional IgG. Purified control antibody freshly spiked in human serum will be used as a reference.
  • a panel of eight combinations of 2 different heavy chains was selected from Table 15 for further analyses. These eight combinations included combinations 3, 4, 5, 6, 9, 10, 11 and 12 (Table 15). In this study, these eight combinations were analyzed, with a strong focus on stability of the Fc part of the IgG.
  • wildtype bispecifics i.e. without CH3 mutations
  • bispecifics based on previously reported CH3 charge mutations were included. Note that for wildtype bispecifics, 2 heavy chains and the common light chain are co-expressed without means for preferential steering towards heterodimers. These 'wildtype bispecifics' thus represent a mixture of AA, AB and BB. All bispecifics in this study were designed to carry the same VH/VL-combinations, ensuring that the observed effects are caused by mutations in the Fc-part of the molecule and not by variation(s) in the Fab parts.
  • bispecific molecules from combinations 3-6 and 9-12 also demonstrated a reduced thermal stability as compared to wildtype. Remarkably, three combinations, however, demonstrated an improved stability as compared to the control CH3 engineered bispecific antibody. Bispecifics of combinations 9, 10 and 11 are
  • bispecific molecules from combinations 3-6 and 9-12 (Table 15), as well as wildtype bispecifics and bispecific molecules obtained when using constructs 1 and 2 (E356K:D399K/ K392D':K409D' combination (charge reversal pair)) were exposed to ten subsequent freeze-thaw cycles by putting the samples at -80°C for at least 15 minutes until they were completely frozen. Thereafter, samples were thawed at room temperature. When they were completely thawed, the freeze-thaw cycle was repeated.
  • K392D':K409D' combination charge reversal pair
  • All IgGs were diluted to 0.2 mg/ml and several stress conditions (2 days at 50°C, 2 weeks at 40°C, 5x freeze-thawing) were applied, aiming to be able to discriminate between the different samples.
  • these high stress levels resulted in conditions in which one of the parental antibodies (the BB parental, carrying two 1122 Fabs) as used in all bispecifics became unstable.
  • aggregation of this protein was detected by UV absorbance. This suggested that this stress condition may not differentiate between instability of the Fab and the CH3 in the bispecific and data resulting from the 50°C incubation should be used cautiously.
  • Nile Red 'Nile Red particles' in Table 16
  • UV spectrometry at 350 nm ('UV 350 nm'); a change in absorption at wavelengths > 320 nm gives information about the aggregation state of the protein.
  • 1,8-ANS fluorescence spectroscopy 1,8-ANS fluorescence spectroscopy; 1,8-ANS binds through electrostatic interactions to cationic groups through ion pair formation and changes in protein structure and/or conformation can be detected
  • UV-Vis absorbance spectra were measured at 25°C with a double beam, two monochromators Cary 300 Bio spectrophotometer from Varian in different quartz cuvettes (such as black low volume Hellma cuvettes with a pathlength of 1.0 cm and clear Hellma cuvettes of 0.2 cm x 1.0 cm). The spectra were monitored between 220 and 450 nm using a pathlength of 1.0 cm. The absorbance around 280 nm provides information on the protein concentration. The region between 320 nm and 450 nm can provide information on the aggregation state of the samples.
  • the 90° light-scattering spectral method was developed to study protein aggregation and was performed as described in Capelle, 2005; Demeule, 2007a.
  • 90° light- scattering spectra were monitored at 25°C using a FluoroMax spectrofluorimeter (Spex, Instruments S.A., Inc. U.K.) by running a synchronous SCan ( em— ⁇ ) between 400 nm and 750 nm with an integration time of 0.01 s.
  • Different slits settings were tested in order to find the optimal conditions. After optimization, the same slit settings were used for all measurements.
  • the fluorescence emission of tryptophan, tyrosine and phenylalanine residues gives information on the local environment of these fluorophores. Changes or differences in hydrophobicity and/or rigidity are measured. Typically, a more hydrophobic and rigid environment leads to an increase in the fluorescence intensity and a blue shift of the emission maximum. Intrinsic fluorescence spectroscopy can provide information on the current state of the protein and monitor changes in the physical and chemical properties. More information on the fluorescence of tyrosine and tryptophan can be found in the book of Lakowicz [Lakowicz, 2006].
  • the fluorescence emission and excitation spectra were recorded at 25°C in different quartz cuvettes. The samples were excited at different wavelengths. Integration times and slit settings were optimized. After optimization, the same integration times and slit settings were applied for all samples.
  • Nile Red staining method was developed to visualize protein aggregates and was performed as described in Demeule et al., 2007b.
  • the microscopy observations were performed on a Leica DM RXE microscope (Leica Microsystems GmbH, Wetzlar, Germany) equipped with a mercury lamp. The images were acquired with a Sony NEX-5 camera and its firmware. The objectives were lOx, 20x and 40x. For microscopy investigations slides with a fixed distance of 0.1 mm between the slide and the cover glass were used. The size of the 4x4 grids is 1 mm x 1 mm and corresponds to 0.1 ⁇ .
  • l-anilinonaphthalene-8-sulfonic acid (1,8-ANS) is an uncharged small hydrophobic fluorescent probe (Mw 299.34 Da) used to study both membrane surfaces and proteins.
  • 1,8-ANS is essentially non-fluorescent in water and only becomes appreciably fluorescent when bound to membranes (quantum yields -0.25) or proteins (quantum yields -0.7). This property of 1,8-ANS makes it a sensitive indicator of protein folding, 5 conformational changes and other processes that modify the exposure of the probe to water. References on 1,8-ANS can be found on the Internet home page of Molecular Probes, www.probes.com.
  • the fluorescence emission spectra of 1,8-ANS were recorded using a FluoroMax spectrometer. A direct comparison of the 1,8-ANS fluorescence between IgGs will not 10 be performed. Each IgG can have different number of 1,8-ANS binding sites and can therefore not be compared. In principle, the lower the 1,8-ANS fluorescence, the less 1,8-ANS molecules are bound to the antibody. The changes in the 1,8-ANS
  • Table 16 Overview of the different forced degradation results on various IgG samples after dilution to 0.2 mg/ml.
  • the most discriminating factors for stability between the proteins are the freeze-thaw cycles and increased temperature. Taking into account the very stringent stress factor of incubating at 50°C, the T366K/L351E,Y349E (combi.#4) and
  • T366K,L351K/L351D,Y349E (combi.#ll) variants are the two most stable proteins 10 within panel, closely followed by T366K,L351K/L351D,Y349D (combi.#10) and
  • Example 21 Native MS on ratio experiments; transfection ratio's from 1:5 to
  • Vectors I-V Previously used antibody VH regions with known ability to pair with the common light chain IGKVl-39 were used for recloning into constructs 1, 2, 68 and 69, resulting in vectors I-V of Table 17.
  • Vectors I-V each containing nucleic acid sequences encoding the common human light chain as well as an Ig heavy chain with different CH3 region and different antigen specificity, were subsequently transfected into cells with different transfection ratios as indicated in Table 18. Results are shown in figure 23.
  • Figures 23A and B show that for the DEKK combination of mutations, when an excess of A or C is present (A or C are on the 'DE side' and B is on the 'KK side'), AB or BC is formed but the surplus of A or C is present as a mixture of both homodimers and half bodies in all cases. However, when an excess of B is present (B is on the 'KK side' and A or C are on the 'DE side'), there is a clear difference. AB or BC is still formed but the surplus of B is essentially absent as homodimer and only half bodies are formed.
  • Figure 23C shows that for the E356K:D399K/K392D':K409D' combination of mutations when an excess of A is present (A is on the 'K392D:K409D side'), the surplus of A is present as a mixture of both homodimers and half bodies in all cases, but also when an excess of B is present (B is on the 'E356K:D399K side'), the surplus of B is present as a mixture of both homodimers and half bodies in all cases. Even at the higher ratios 1:3 and 1:5 no half bodies B are observed although homodimers are present, indicating that the
  • E356K:D399K side does not disfavour homodimerization as much as the KK-side of the DEKK combination.
  • the DEKK combination of mutations offers a clear benefit over the charge reversal CH3 mutations, in that one of the chains of the heterodimer does not form homodimers.
  • the IgG samples were further tested in a sandwich ELISA to confirm the functional presence of the desired specificities. Coating of ELISA plates was done with fibrinogen or thyroglobulin and detection was performed with fluorescein-labelled thyroglobulin or - tetanus toxoid. The detection antigens were labelled with fluorescein (Pierce NHS-fluorescin Antibody Labeling kit, cat. # 53029) according to the manufacturer's instructions. Fluorescein-labeled antigens could subsequently be detected by a FITC- conjugated anti-fluorescein antibody (Roche diagnostics, cat. # 11426346910).
  • results of the bispecific ELISA are summarized in Table 21.
  • the greyed cells indicate the expected species for each transfection. Generally, the results meet the expected outcome with view exceptions as indicated in italic or bold.
  • BC tr. #1 and 2
  • AC tr.# 3
  • results of bispecific ELISA may suffer from high background levels. These background levels may also be caused by the potential presence of half-bodies in the sample.
  • results of bispecific ELISA indeed confirmed that an error had occurred in transfection #11, as the species AC (bold value) was detected rather than BC.
  • Table 21 OD450 values from bispecific ELISA
  • Example 23 improved mixtures of two bispecific antibodies recognizing 4 different epitopes (AB and CD) from a single cell
  • CH3 domain-comprising polypeptide chains comprising these novel charge pair mutants can be expressed in cells together with previously known charge-engineered CH3 domain-comprising polypeptide chains or potentially with SEED bodies, and are likely to result in the preferential formation of two bispecific molecules only. From the above examples it was clear that the DEKK combination of mutations is excellent for the production of one bispecific (AB) or two bispecifics (AB plus AC) by clonal cells where dimerization of the heavy chains is driven by the CH3 domains.
  • Table 22 CH3 variants analyzed in HADDOCK, with one letter codes for assigned for each CH3-variant carrying heavy chain.
  • *Wildtype chains are designated 'C and 'D' for matters of consistency; **The charge reversal variants are designated ⁇ and B' when combined with knob-into-hole variants, and are designated 'C and D' when combined with DE/KK variants.
  • Figure 25 shows that, based on these HADDOCK predictions, combining the CH3 combinations of DEKK with charge reversal CH3 combinations is most likely to be successful in forming the desired combination of two bispecifics (AB and CD) without contaminating by-products (especially AC, AD, BC, BD) when co-transfected in a single cell.
  • these undesired bispecific species AC, AD, BC, and BD have relatively high HADDOCK scores, whereas the desired AB and CD species have the lowest HADDOCK scores.
  • the CH3 combinations of DEKK or charge reversal will be put into a construct carrying the same specificity (e.g.
  • the mass difference between the (expected) species has to be sufficient, i.e. > 190 Da.
  • Four individual VHs have been selected and the masses of these were such that the expected species upon co-transfection could be identified and separated by nMS.
  • the mass differences between the 4 selected VHs are also large enough to identify most of the possible contaminants in the mixtures, in addition to the two desired species. Selected VHs are listed in Table 23.
  • Table 25 provides a further overview of masses of the expected species, and possible contaminants, of transfections # 9- 11 of Table 24.
  • Table 25 For each of transfections # 9- 11, the species are sorted by mass, mass difference is calculated with the mass above. Grey cells: expected (and desired) species; italics: mass difference too small to separate in nMS analysis. *Species letters represent half -bodies; two-letter code intact IgG.
  • transfections # 3 and # 4 resulted in the expected mismatch between 'KK' constructs and either 'E356K:D399K or 'K392D:K409D' and the amount of half bodies in protein samples from these transfections exceeded the amount of full IgG molecules.
  • Transfections # 7 and # 8 resulted in protein samples wherein both half bodies and full IgG is present in about equal amounts.
  • the three IgG batches included 1) wildtype anti-tetanus toxoid parental antibody 1337: 1337 (two MF1337 Fabs on a wildtype Fc backbone); 2) wildtype anti-tetanus toxoid parental antibody 1516: 1516 (two MF1516 Fabs on a wildtype Fc backbone); 3) CH3 engineered bispecific anti-tetanus toxoid antibody 1516: 1337 that carries the DEKK combination of mutations in its Fc region (MF1516 Fab on DE-side, MF1337 Fab on KK-side).
  • the parental antibodies 1337: 1337 and 1516: 1516 were chosen as specificities to be included in the DEKK-bispecific product, as it was known based on previous studies that no pre-dose serum response against these antibodies was present in several mice strains. NB: the presence of a pre-dose serum response would of course invalidate the study.
  • the three IgG batches were prepared as previously described, but the DNA used for transfection was made using an endo-free maxiprep kit to ensure that the amount of endotoxins is as low as possible.
  • the batches were subsequently tested for protein concentration, aggregate levels, endotoxin levels and percentage bispecific product. It was demonstrated that the acceptance criteria for subsequent use of the IgG batches in a pK study were met, i.e. the IgG concentration after gel filtration was >0.3 mg/ml, aggregate levels were ⁇ 5%, endotoxin levels were ⁇ 3 EU/mg protein and the DEKK batch contained > 90% bispecific IgG.
  • mice Female C57BL/6J mice (Harlan, The Netherlands) were dosed at 1 mg/kg human IgG (5 ml/kg immunoglobulin solution/kg body weight). At dosing time, the animals were between 7-8 weeks of age and had a body weight of about 18-20 grams. Blood samples were collected pre-dose and at 15, 60 minutes, and 2, 4, 8, 24, 48, 96, 168, 268 and 336 h after dosing. Serum samples were prepared and stored at ⁇ -20 °C until analysis. Each group consisted of 3 subgroups of 4 mice, i.e. 12 mice/group. From each mice 6 time points were sampled. The welfare of the animals was maintained in accordance with the general principles governing the use of animals in experiments of the European Communities (Directive 86/609/EEC) and Dutch legislation (The
  • mice of Group 1 received the full length monospecific IgG 1516: 1516 antibody
  • mice of Group 2 received the full length monospecific IgG 1337: 1337 antibody (squares); Mice of Group 3 received the full length bispecific IgG 1516: 1337 antibody (diamonds), with DEKK engineered CH3 regions (1516 on the DE-side and 1337 on the KK-side).
  • An ELISA assay was applied for the quantitative analysis of monoclonal human antibodies in mouse serum using a quantitative human IgG ELISA (ZeptoMetrix, NY USA; ELISA kit nr. 0801182). Briefly, the ELISA assay is based on the principle that the human monoclonal antibody binds to anti-human IgG coated in a 96-wells ELISA plate.
  • Bound antibody was subsequently visualized using a polyclonal antihuman IgG antibody conjugated with horseradish peroxidase (HRP).
  • HRP horseradish peroxidase
  • OD optical density

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DK13720614.0T DK2838917T3 (da) 2012-04-20 2013-04-19 Fremgangsmåder og midler til frembringelse af heterodimere ig-lignende molekyler
CN202510431296.0A CN120383672A (zh) 2012-04-20 2013-04-19 用于产生免疫球蛋白样分子的方法和手段
MX2014012700A MX360109B (es) 2012-04-20 2013-04-19 Metodos y medios para la produccion de moleculas de tipo ig.
SG11201406770WA SG11201406770WA (en) 2012-04-20 2013-04-19 Methods and means for the production of ig-like molecules
NZ630551A NZ630551A (en) 2012-04-20 2013-04-19 Methods and means for the production of ig-like molecules
EP19177410.8A EP3594232A1 (en) 2012-04-20 2013-04-19 Methods and means for the production of ig-like molecules
SM20190493T SMT201900493T1 (it) 2012-04-20 2013-04-19 Metodi e mezzi per la produzione di molecole simili a ig eterodimeriche
HRP20191446 HRP20191446T1 (hr) 2012-04-20 2013-04-19 Postupci i sredstva za proizvodnju heterodimernih ig-sličnih molekula
CA2871068A CA2871068C (en) 2012-04-20 2013-04-19 Methods and means for the production of ig-like molecules
CN202111365429.7A CN114163530B (zh) 2012-04-20 2013-04-19 用于产生免疫球蛋白样分子的方法和手段
ES13720614T ES2743399T3 (es) 2012-04-20 2013-04-19 Métodos y medios para la producción de moléculas heterodiméricas similares a Ig
SI201331553T SI2838917T1 (sl) 2012-04-20 2013-04-19 Postopki in sredstva za produkcijo heterodimernih IG-podobnih molekul
RSP20191034 RS59263B1 (sr) 2012-04-20 2013-04-19 Postupci i sredstva za proizvodnju heterodimernih ig-sličnih molekula
KR1020147032231A KR102104601B1 (ko) 2012-04-20 2013-04-19 Ig-유사 분자의 제조방법 및 제조수단
US14/395,325 US20150196637A1 (en) 2012-04-20 2013-04-19 Methods and means for the production of ig-like molecules
JP2015506927A JP6272299B2 (ja) 2012-04-20 2013-04-19 Ｉｇ様分子を生産する方法、Ｉｇ様分子の混合物、組み換え宿主細胞、医薬組成物、宿主細胞を作成する方法、および培養物
CN201380032190.9A CN104520319B (zh) 2012-04-20 2013-04-19 用于产生免疫球蛋白样分子的方法和手段
EP13720614.0A EP2838917B1 (en) 2012-04-20 2013-04-19 Methods and means for the production of heterodimeric ig-like molecules
EA201491909A EA035344B1 (ru) 2012-04-20 2013-04-19 Способ получения двух антител из одной клетки-хозяина
PL13720614T PL2838917T3 (pl) 2012-04-20 2013-04-19 Sposoby i środki do wytwarzania heterodimerycznych cząsteczek podobnych do Ig
AU2013249985A AU2013249985B2 (en) 2012-04-20 2013-04-19 Methods and means for the production of Ig-like molecules
KR1020207011173A KR102382304B1 (ko) 2012-04-20 2013-04-19 Ig-유사 분자의 제조방법 및 제조수단
HK15106743.0A HK1206041B (en) 2012-04-20 2013-04-19 Methods and means for the production of heterodimeric ig-like molecules
LTEP13720614.0T LT2838917T (lt) 2012-04-20 2013-04-19 Heterodimerinių ig tipo molekulių gamybos būdai ir priemonės
MX2024012547A MX2024012547A (es) 2012-04-20 2014-10-20 Metodos y medios para la produccion de moleculas de tipo ig
MX2024012548A MX2024012548A (es) 2012-04-20 2014-10-20 Metodos y medios para la produccion de moleculas de tipo ig
ZA2014/07835A ZA201407835B (en) 2012-04-20 2014-10-27 Methods an means for the production of ig-like molecules
AU2018201325A AU2018201325B2 (en) 2012-04-20 2018-02-23 Methods and means for the production of Ig-like molecules
CY20191100914T CY1122118T1 (el) 2012-04-20 2019-08-28 Μεθοδοι και μεσα για την παραγωγη των ετεροδιμερων μοριων τυπου ig

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Cited By (241)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014177461A1 (en) 2013-04-29 2014-11-06 F. Hoffmann-La Roche Ag Fcrn-binding abolished anti-igf-1r antibodies and their use in the treatment of vascular eye diseases
WO2014177460A1 (en) 2013-04-29 2014-11-06 F. Hoffmann-La Roche Ag Human fcrn-binding modified antibodies and methods of use
US20140378665A1 (en) * 2012-01-26 2014-12-25 Amgen Inc. Growth differentiation factor 15 (gdf-15) polypeptides
WO2015101586A1 (en) 2014-01-03 2015-07-09 F. Hoffmann-La Roche Ag Bispecific anti-hapten/anti-blood brain barrier receptor antibodies, complexes thereof and their use as blood brain barrier shuttles
WO2015101588A1 (en) 2014-01-06 2015-07-09 F. Hoffmann-La Roche Ag Monovalent blood brain barrier shuttle modules
WO2015107026A1 (en) 2014-01-15 2015-07-23 F. Hoffmann-La Roche Ag Fc-region variants with modified fcrn- and maintained protein a-binding properties
WO2015150446A1 (en) 2014-04-02 2015-10-08 F. Hoffmann-La Roche Ag Method for detecting multispecific antibody light chain mispairing
WO2015150447A1 (en) 2014-04-02 2015-10-08 F. Hoffmann-La Roche Ag Multispecific antibodies
EP2982694A1 (en) 2014-08-04 2016-02-10 EngMab AG Bispecific antibodies against cd3epsilon and bcma
WO2016055592A1 (en) 2014-10-09 2016-04-14 Engmab Ag Bispecific antibodies against cd3epsilon and ror1
EP3023437A1 (en) 2014-11-20 2016-05-25 EngMab AG Bispecific antibodies against CD3epsilon and BCMA
US20160168213A1 (en) * 2013-07-31 2016-06-16 Amgen Inc. Growth differentiation factor 15 (gdf-15) constructs
WO2017009258A1 (en) 2015-07-10 2017-01-19 Genmab A/S Axl-specific antibody-drug conjugates for cancer treatment
WO2017034770A1 (en) 2015-08-26 2017-03-02 Bison Therapeutics Inc. Multispecific antibody platform and related methods
EP3150637A1 (en) 2015-10-02 2017-04-05 F. Hoffmann-La Roche AG Multispecific antibodies
WO2017055391A1 (en) 2015-10-02 2017-04-06 F. Hoffmann-La Roche Ag Bispecific t cell activating antigen binding molecules binding mesothelin and cd3
WO2017055389A1 (en) 2015-10-02 2017-04-06 F. Hoffmann-La Roche Ag Bispecific anti-ceaxcd3 t cell activating antigen binding molecules
WO2017055314A1 (en) 2015-10-02 2017-04-06 F. Hoffmann-La Roche Ag Bispecific anti-cd19xcd3 t cell activating antigen binding molecules
WO2017055399A1 (en) 2015-10-02 2017-04-06 F. Hoffmann-La Roche Ag Cellular based fret assay for the determination of simultaneous binding
WO2017055404A1 (en) 2015-10-02 2017-04-06 F. Hoffmann-La Roche Ag Bispecific antibodies specific for pd1 and tim3
WO2017055395A1 (en) 2015-10-02 2017-04-06 F. Hoffmann-La Roche Ag Anti-cd3xrob04 bispecific t cell activating antigen binding molecules
WO2017055392A1 (en) 2015-10-02 2017-04-06 F. Hoffmann-La Roche Ag Anti-cd3xcd44v6 bispecific t cell activating antigen binding molecules
WO2017055385A1 (en) 2015-10-02 2017-04-06 F. Hoffmann-La Roche Ag Anti-cd3xgd2 bispecific t cell activating antigen binding molecules
WO2017055388A2 (en) 2015-10-02 2017-04-06 F. Hoffmann-La Roche Ag Bispecific t cell activating antigen binding molecules
WO2017055393A1 (en) 2015-10-02 2017-04-06 F. Hoffmann-La Roche Ag Anti-cd3xtim-3 bispecific t cell activating antigen binding molecules
WO2017072208A1 (en) * 2015-10-29 2017-05-04 F. Hoffmann-La Roche Ag Transgenic rabbit with common light chain
US9676845B2 (en) 2009-06-16 2017-06-13 Hoffmann-La Roche, Inc. Bispecific antigen binding proteins
EP3178848A1 (en) 2015-12-09 2017-06-14 F. Hoffmann-La Roche AG Type ii anti-cd20 antibody for reducing formation of anti-drug antibodies
WO2017186853A1 (en) * 2016-04-27 2017-11-02 Bcrt Holding Bv T cell-based bi-specific immunotherapeutics
WO2017191101A1 (en) 2016-05-02 2017-11-09 F. Hoffmann-La Roche Ag The contorsbody - a single chain target binder
EP3252078A1 (en) 2016-06-02 2017-12-06 F. Hoffmann-La Roche AG Type ii anti-cd20 antibody and anti-cd20/cd3 bispecific antibody for treatment of cancer
WO2018007314A1 (en) 2016-07-04 2018-01-11 F. Hoffmann-La Roche Ag Novel antibody format
WO2018011421A1 (en) 2016-07-14 2018-01-18 Genmab A/S Multispecific antibodies against cd40 and cd137
US9890204B2 (en) 2009-04-07 2018-02-13 Hoffmann-La Roche Inc. Trivalent, bispecific antibodies
US9914776B2 (en) 2014-08-04 2018-03-13 Hoffmann-La Roche Inc. Bispecific T cell activating antigen binding molecules
WO2018059502A1 (en) * 2016-09-29 2018-04-05 Beijing Hanmi Pharmaceutical Co., Ltd. Heterodimeric immunoglobulin constructs and preparation methods thereof
WO2018060301A1 (en) 2016-09-30 2018-04-05 F. Hoffmann-La Roche Ag Bispecific antibodies against cd3
WO2018083126A1 (en) 2016-11-01 2018-05-11 Genmab B.V. Polypeptide variants and uses thereof
US9982036B2 (en) 2011-02-28 2018-05-29 Hoffmann-La Roche Inc. Dual FC antigen binding proteins
US9994646B2 (en) 2009-09-16 2018-06-12 Genentech, Inc. Coiled coil and/or tether containing protein complexes and uses thereof
EP3274457A4 (en) * 2015-03-26 2018-08-08 Acceleron Pharma Inc. Follistatin-related fusion proteins and uses thereof
WO2018146317A1 (en) 2017-02-10 2018-08-16 Genmab B.V. Polypeptide variants and uses thereof
WO2018162749A1 (en) 2017-03-09 2018-09-13 Genmab A/S Antibodies against pd-l1
US10087250B2 (en) 2012-10-08 2018-10-02 Roche Glycart Ag Fc-free antibodies comprising two fab-fragments and methods of use
WO2018178396A1 (en) 2017-03-31 2018-10-04 Genmab Holding B.V. Bispecific anti-cd37 antibodies, monoclonal anti-cd37 antibodies and methods of use thereof
WO2018184964A1 (en) 2017-04-03 2018-10-11 F. Hoffmann-La Roche Ag Immunoconjugates of an anti-pd-1 antibody with a mutant il-2 or with il-15
WO2018184966A1 (en) 2017-04-03 2018-10-11 F. Hoffmann-La Roche Ag Antibodies binding to steap-1
WO2018185043A1 (en) 2017-04-05 2018-10-11 F. Hoffmann-La Roche Ag Bispecific antibodies specifically binding to pd1 and lag3
WO2018184965A1 (en) 2017-04-03 2018-10-11 F. Hoffmann-La Roche Ag Immunoconjugates of il-2 with an anti-pd-1 and tim-3 bispecific antibody
WO2018189220A1 (en) 2017-04-13 2018-10-18 F. Hoffmann-La Roche Ag An interleukin-2 immunoconjugate, a cd40 agonist, and optionally a pd-1 axis binding antagonist for use in methods of treating cancer
US10106600B2 (en) 2010-03-26 2018-10-23 Roche Glycart Ag Bispecific antibodies
US10138293B2 (en) 2007-12-21 2018-11-27 Hoffmann-La Roche, Inc. Bivalent, bispecific antibodies
WO2018220099A1 (en) 2017-06-02 2018-12-06 F. Hoffmann-La Roche Ag Type ii anti-cd20 antibody and anti-cd20/cd3 bispecific antibody for treatment of cancer
WO2018224609A1 (en) 2017-06-07 2018-12-13 Genmab B.V. Therapeutic antibodies based on mutated igg hexamers
US10155815B2 (en) 2013-02-26 2018-12-18 Roche Glycart Ag Bispecific T cell activating antigen binding molecules
WO2019009727A1 (en) 2017-07-06 2019-01-10 Merus N.V. ANTI-PD1 ANTI-TIM3 BISPECIFIC ANTIBODIES
WO2019009728A1 (en) 2017-07-06 2019-01-10 Merus N.V. ANTIBODIES THAT MODULATE A BIOLOGICAL ACTIVITY EXPRESSED BY A CELL
WO2019009726A1 (en) 2017-07-06 2019-01-10 Merus N.V. BINDING MOLECULES FOR MODULATING THE BIOLOGICAL ACTIVITY EXPRESSED BY A CELL
WO2019025545A1 (en) 2017-08-04 2019-02-07 Genmab A/S BINDING AGENTS BINDING TO PD-L1 AND CD137 AND THEIR USE
WO2019031965A1 (en) 2017-08-09 2019-02-14 Merus N.V. ANTIBODIES THAT BIND EGFR AND CMET
WO2018075692A3 (en) * 2016-10-19 2019-04-04 Invenra Inc. Antibody constructs
US10251952B2 (en) 2014-06-26 2019-04-09 Hoffmann-La Roche Inc. Humanized anti-tau(pS422) antibody brain shuttles and use thereof
EP3463451A1 (en) * 2016-05-26 2019-04-10 Qilu Puget Sound Biotherapeutics Corporation Mixtures of antibodies
WO2019070047A1 (ja) 2017-10-06 2019-04-11 小野薬品工業株式会社 二重特異性抗体
WO2019077092A1 (en) 2017-10-20 2019-04-25 F. Hoffmann-La Roche Ag METHOD FOR GENERATING MULTISPECIFIC ANTIBODIES FROM MONOSPECIFIC ANTIBODIES
WO2019086394A1 (en) 2017-11-01 2019-05-09 F. Hoffmann-La Roche Ag The compbody - a multivalent target binder
WO2019086362A1 (en) 2017-10-30 2019-05-09 F. Hoffmann-La Roche Ag Method for in vivo generation of multispecific antibodies from monospecific antibodies
WO2019086395A1 (en) 2017-11-01 2019-05-09 F. Hoffmann-La Roche Ag Trifab-contorsbody
US10323099B2 (en) 2013-10-11 2019-06-18 Hoffmann-La Roche Inc. Multispecific domain exchanged common variable light chain antibodies
WO2019122052A2 (en) 2017-12-21 2019-06-27 F. Hoffmann-La Roche Ag Antibodies binding to hla-a2/wt1
WO2019145455A1 (en) 2018-01-24 2019-08-01 Genmab B.V. Polypeptide variants and uses thereof
WO2019154776A1 (en) 2018-02-06 2019-08-15 F. Hoffmann-La Roche Ag Treatment of ophthalmologic diseases
WO2019154890A1 (en) 2018-02-09 2019-08-15 F. Hoffmann-La Roche Ag Antibodies binding to gprc5d
WO2019156199A1 (ja) 2018-02-09 2019-08-15 小野薬品工業株式会社 二重特異性抗体
WO2019190327A2 (en) 2018-03-30 2019-10-03 Merus N.V. Multivalent antibody
WO2019202041A1 (en) 2018-04-18 2019-10-24 F. Hoffmann-La Roche Ag Multispecific antibodies and use thereof
US10465000B2 (en) 2013-12-20 2019-11-05 Hoffmann-La Roche Inc. Humanized anti-Tau(pS422) antibodies and methods of use
WO2019211472A1 (en) 2018-05-03 2019-11-07 Genmab B.V. Antibody variant combinations and uses thereof
WO2019232484A1 (en) 2018-06-01 2019-12-05 Compugen Ltd Anti-pvrig/anti-tigit bispecific antibodies and methods of use
WO2019243636A1 (en) 2018-06-22 2019-12-26 Genmab Holding B.V. Anti-cd37 antibodies and anti-cd20 antibodies, compositions and methods of use thereof
WO2020012038A1 (en) 2018-07-13 2020-01-16 Genmab A/S Trogocytosis-mediated therapy using cd38 antibodies
WO2020012036A1 (en) 2018-07-13 2020-01-16 Genmab A/S Variants of cd38 antibody and uses thereof
EP3611188A1 (en) 2014-11-06 2020-02-19 F. Hoffmann-La Roche AG Fc-region variants with modified fcrn-binding and methods of use
US10596257B2 (en) 2016-01-08 2020-03-24 Hoffmann-La Roche Inc. Methods of treating CEA-positive cancers using PD-1 axis binding antagonists and anti-CEA/anti-CD3 bispecific antibodies
WO2020060405A1 (en) 2018-09-19 2020-03-26 Lava Therapeutics B.V. Dual acting cd1d immunoglobulin
US10611825B2 (en) 2011-02-28 2020-04-07 Hoffmann La-Roche Inc. Monovalent antigen binding proteins
WO2020070313A1 (en) 2018-10-04 2020-04-09 Genmab Holding B.V. Pharmaceutical compositions comprising bispecific anti-cd37 antibodies
US10633457B2 (en) 2014-12-03 2020-04-28 Hoffmann-La Roche Inc. Multispecific antibodies
WO2020089051A1 (en) 2018-10-29 2020-05-07 F. Hoffmann-La Roche Ag Antibody formulation
WO2020094744A1 (en) 2018-11-06 2020-05-14 Genmab A/S Antibody formulation
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WO2020136060A1 (en) 2018-12-28 2020-07-02 F. Hoffmann-La Roche Ag A peptide-mhc-i-antibody fusion protein for therapeutic use in a patient with amplified immune response
WO2020141974A1 (en) 2018-12-31 2020-07-09 Merus N.V. Truncated multivalent multimers
WO2020141973A1 (en) 2018-12-31 2020-07-09 Merus N.V. Mixed binding domains
WO2020159368A1 (en) 2019-02-01 2020-08-06 Lava Therapeutics B.V. Novel cd40-binding antibodies
NL2022494B1 (en) 2019-02-01 2020-08-19 Lava Therapeutics B V Novel CD40-binding antibodies
US10752664B2 (en) 2011-04-08 2020-08-25 Amgen Inc. Method of treating or ameliorating metabolic disorders using growth differentiation factor 15 (GDF-15)
US10781262B2 (en) 2014-11-20 2020-09-22 Hoffmann-La Roche Inc. Combination therapy of T cell activating bispecific antigen binding molecules and PD-1 axis binding antagonists
WO2020204708A1 (en) 2019-03-29 2020-10-08 Merus N.V. Cd3 binding molecules
US10822402B2 (en) 2015-06-24 2020-11-03 Hoffmann-La Roche Inc. Humanized anti-tau(pS422) antibodies and methods of use
WO2020225456A1 (en) 2019-05-09 2020-11-12 Genmab B.V. Dosage regimens for a combination of anti-dr5 antibodies for use in treating cancer
US10844127B2 (en) 2014-02-28 2020-11-24 Merus N.V. Antibodies that bind EGFR and ErbB3
WO2021001289A1 (en) 2019-07-02 2021-01-07 F. Hoffmann-La Roche Ag Immunoconjugates comprising a mutant interleukin-2 and an anti-cd8 antibody
WO2021006199A1 (ja) 2019-07-05 2021-01-14 小野薬品工業株式会社 Ｐｄ－１／ｃｄ３二重特異性タンパク質による血液がん治療
WO2021009146A1 (en) 2019-07-15 2021-01-21 F. Hoffmann-La Roche Ag Antibodies binding to nkg2d
WO2021018925A1 (en) 2019-07-31 2021-02-04 F. Hoffmann-La Roche Ag Antibodies binding to gprc5d
WO2021020416A1 (ja) 2019-07-30 2021-02-04 小野薬品工業株式会社 二重特異性抗体
WO2021018859A2 (en) 2019-07-31 2021-02-04 F. Hoffmann-La Roche Ag Antibodies binding to gprc5d
WO2021025140A1 (ja) 2019-08-08 2021-02-11 小野薬品工業株式会社 二重特異性タンパク質
WO2021034194A2 (en) 2019-08-19 2021-02-25 Merus N.V. Treatment of cancer with a combination of an antibody that binds lgr5 and egfr and a topoisomerase i inhibitor.
EP3792283A1 (en) 2019-09-16 2021-03-17 Lava Therapeutics B.V. Treatment of cancer comprising administration of vgamma9vdelta2 t cell receptor binding antibodies
EP3638700A4 (en) * 2017-06-14 2021-04-21 Dingfu Biotarget Co., Ltd HETERODIMER PROTEIN AND USE OF IT
WO2021089850A1 (en) 2019-11-06 2021-05-14 Genmab B.V. Antibody variant combinations and uses thereof
US11013801B2 (en) 2015-12-09 2021-05-25 Hoffmann-La Roche Inc. Treatment method
WO2021122733A1 (en) 2019-12-18 2021-06-24 F. Hoffmann-La Roche Ag Bispecific anti-ccl2 antibodies
WO2021122875A1 (en) 2019-12-18 2021-06-24 F. Hoffmann-La Roche Ag Antibodies binding to hla-a2/mage-a4
EP3842453A1 (en) 2014-11-06 2021-06-30 F. Hoffmann-La Roche AG Fc-region variants with modified fcrn- and protein a-binding properties
WO2021144457A1 (en) 2020-01-16 2021-07-22 Genmab A/S Formulations of cd38 antibodies and uses thereof
WO2021154073A1 (en) 2020-01-29 2021-08-05 Merus N.V. Means and method for modulating immune cell engaging effects.
WO2021156258A1 (en) 2020-02-04 2021-08-12 BioNTech SE Treatment involving antigen vaccination and binding agents binding to pd-l1 and cd137
EP3878866A1 (en) 2013-04-29 2021-09-15 F. Hoffmann-La Roche AG Fc-receptor binding modified asymmetric antibodies and methods of use
WO2021185934A1 (en) 2020-03-18 2021-09-23 Genmab A/S Antibodies binding to b7h4
WO2021209402A2 (en) 2020-04-15 2021-10-21 F. Hoffmann-La Roche Ag Immunoconjugates
WO2021215926A1 (en) 2020-04-24 2021-10-28 Merus N.V. Treatment of cancers with an antibody that binds lgr5 and egfr
US11161889B2 (en) 2018-04-09 2021-11-02 Amgen Inc. Growth differentiation factor 15 fusion proteins
WO2021222552A1 (en) 2020-04-30 2021-11-04 Celgene Corporation Methods of treating cytokine-related adverse events
EP3912998A2 (en) 2015-10-23 2021-11-24 Merus N.V. Binding molecules that inhibit cancer growth
WO2021235936A1 (en) 2020-05-21 2021-11-25 Merus N.V. Methods and means for the production of ig-like molecules
WO2021255143A1 (en) 2020-06-19 2021-12-23 F. Hoffmann-La Roche Ag Antibodies binding to cd3 and folr1
WO2021255138A1 (en) 2020-06-19 2021-12-23 F. Hoffmann-La Roche Ag Immune activating fc domain binding molecules
WO2021255142A1 (en) 2020-06-19 2021-12-23 F. Hoffmann-La Roche Ag Antibodies binding to cd3
WO2021255146A1 (en) 2020-06-19 2021-12-23 F. Hoffmann-La Roche Ag Antibodies binding to cd3 and cea
WO2021255155A1 (en) 2020-06-19 2021-12-23 F. Hoffmann-La Roche Ag Antibodies binding to cd3 and cd19
WO2022008646A1 (en) 2020-07-08 2022-01-13 LAVA Therapeutics N.V. Antibodies that bind psma and gamma-delta t cell receptors
US11230697B2 (en) 2006-09-01 2022-01-25 Therapeutic Human Polyclonals Inc. Enhanced expression of human or humanized immunoglobulin in non-human transgenic animals
WO2022018294A1 (en) 2020-07-23 2022-01-27 Genmab B.V. A combination of anti-dr5 antibodies and an immunomodulatory imide drug for use in treating multiple myeloma
WO2022023559A1 (en) 2020-07-31 2022-02-03 Curevac Ag Nucleic acid encoded antibody mixtures
US11242390B2 (en) 2016-03-22 2022-02-08 Hoffmann-La Roche Inc. Protease-activated T cell bispecific molecules
WO2022029011A1 (en) 2020-08-06 2022-02-10 BioNTech SE Binding agents for coronavirus s protein
US11279770B2 (en) 2014-02-28 2022-03-22 Merus N.V. Antibody that binds ErbB-2 and ErbB-3
US11286300B2 (en) 2015-10-01 2022-03-29 Hoffmann-La Roche Inc. Humanized anti-human CD19 antibodies and methods of use
WO2022069724A1 (en) 2020-10-02 2022-04-07 Genmab A/S Antibodies capable of binding to ror2 and bispecific antibodies binding to ror2 and cd3
US11319378B2 (en) 2016-11-18 2022-05-03 Beijing Hanmi Pharmaceutical Co., Ltd. Anti-PD-1/anti-HER2 natural antibody structural heterodimeric bispecific antibody and method of preparing the same
US11332541B2 (en) 2018-06-09 2022-05-17 Boehringer Ingelheim International Gmbh Multi-specific binding proteins for cancer treatment
US11359015B2 (en) 2015-07-15 2022-06-14 Genmab A/S Humanized or chimeric CD3 antibodies
WO2022122973A1 (en) 2020-12-10 2022-06-16 LAVA Therapeutics N.V. Antibodies that bind gamma-delta t cell receptors
WO2022131912A1 (en) 2020-12-15 2022-06-23 Merus N.V. Treatment of cancers with an antibody that binds lgr5 and egfr
WO2022129120A1 (en) 2020-12-17 2022-06-23 F. Hoffmann-La Roche Ag Anti-hla-g antibodies and use thereof
WO2022148853A1 (en) 2021-01-11 2022-07-14 F. Hoffmann-La Roche Ag Immunoconjugates
WO2022148732A1 (en) 2021-01-06 2022-07-14 F. Hoffmann-La Roche Ag Combination therapy employing a pd1-lag3 bispecific antibody and a cd20 t cell bispecific antibody
WO2022180271A1 (en) 2021-02-26 2022-09-01 LAVA Therapeutics N.V. Antibodies that bind CD123 and gamma-delta T cell receptors
US11459404B2 (en) 2013-02-26 2022-10-04 Roche Glycart Ag Bispecific T cell activating antigen binding molecules
WO2022211625A1 (en) 2021-03-31 2022-10-06 Merus N.V. Novel multispecific antibodies
WO2022228706A1 (en) 2021-04-30 2022-11-03 F. Hoffmann-La Roche Ag Dosing for treatment with anti-cd20/anti-cd3 bispecific antibody
WO2022228705A1 (en) 2021-04-30 2022-11-03 F. Hoffmann-La Roche Ag Dosing for combination treatment with anti-cd20/anti-cd3 bispecific antibody and anti-cd79b antibody drug conjugate
WO2022234146A1 (en) 2021-05-07 2022-11-10 Genmab A/S PHARMACEUTICAL COMPOSITIONS COMPRISING BISPECIFIC ANTIBODIES BINDING TO B7H4 and CD3
WO2022263501A1 (en) 2021-06-18 2022-12-22 F. Hoffmann-La Roche Ag Bispecific anti-ccl2 antibodies
WO2022268740A1 (en) 2021-06-21 2022-12-29 Genmab A/S Combination dosage regime of cd137 and pd-l1 binding agents
WO2023279092A2 (en) 2021-07-02 2023-01-05 Genentech, Inc. Methods and compositions for treating cancer
WO2023001884A1 (en) 2021-07-22 2023-01-26 F. Hoffmann-La Roche Ag Heterodimeric fc domain antibodies
WO2023010095A1 (en) 2021-07-28 2023-02-02 F. Hoffmann-La Roche Ag Methods and compositions for treating cancer
WO2023031473A1 (en) 2021-09-06 2023-03-09 Genmab B.V. Antibodies capable of binding to cd27, variants thereof and uses thereof
WO2023037333A1 (en) 2021-09-13 2023-03-16 Janssen Biotech, Inc CD33 X Vδ2 MULTISPECIFIC ANTIBODIES FOR THE TREATMENT OF CANCER
WO2023059191A1 (en) 2021-10-06 2023-04-13 Merus N.V. Treatment of immune checkpoint inhibitor-treated cancers with high egfr expression using an antibody that binds at least egfr
WO2023057571A1 (en) 2021-10-08 2023-04-13 Genmab A/S Antibodies binding to cd30 and cd3
WO2023062048A1 (en) 2021-10-14 2023-04-20 F. Hoffmann-La Roche Ag Alternative pd1-il7v immunoconjugates for the treatment of cancer
WO2023062050A1 (en) 2021-10-14 2023-04-20 F. Hoffmann-La Roche Ag New interleukin-7 immunoconjugates
WO2023067138A1 (en) 2021-10-21 2023-04-27 LAVA Therapeutics N.V. Uses of gamma delta t cell activating antibodies
US11639397B2 (en) 2011-08-23 2023-05-02 Roche Glycart Ag Bispecific antibodies specific for T-cell activating antigens and a tumor antigen and methods of use
WO2023089083A1 (en) 2021-11-19 2023-05-25 Merus N.V. Multispecific binding moieties comprising pd-1 and tgf-brii binding domains
WO2023094282A1 (en) 2021-11-25 2023-06-01 F. Hoffmann-La Roche Ag Quantification of low amounts of antibody sideproducts
US11673968B2 (en) 2014-06-26 2023-06-13 Hoffmann-La Roche Inc. Anti-BRDU antibodies and methods of use
WO2023104938A1 (en) 2021-12-10 2023-06-15 F. Hoffmann-La Roche Ag Antibodies binding to cd3 and plap
US11685786B2 (en) 2016-09-23 2023-06-27 Merus N.V. Binding molecules that bind CD137 and PD-L1
US11753471B2 (en) 2018-02-08 2023-09-12 Beijing Hanmi Pharmaceutical Co., Ltd. Anti-PD-1/anti-HER2 natural antibody structural heterodimeric bispecific antibody and method of preparing same
WO2023172133A1 (en) 2022-03-07 2023-09-14 Merus N.V. Combination therapy including antibodies that bind egfr and cmet
WO2023172134A1 (en) 2022-03-07 2023-09-14 Merus N.V. Treatment with an antibody that binds egfr and cmet.
WO2023174952A1 (en) 2022-03-15 2023-09-21 Genmab A/S Binding agents binding to epcam and/or cd137
WO2023180353A1 (en) 2022-03-23 2023-09-28 F. Hoffmann-La Roche Ag Combination treatment of an anti-cd20/anti-cd3 bispecific antibody and chemotherapy
US11780925B2 (en) 2017-03-31 2023-10-10 Merus N.V. ErbB-2 and ErbB3 binding bispecific antibodies for use in the treatment of cells that have an NRG1 fusion gene
WO2023198727A1 (en) 2022-04-13 2023-10-19 F. Hoffmann-La Roche Ag Pharmaceutical compositions of anti-cd20/anti-cd3 bispecific antibodies and methods of use
WO2023218051A1 (en) 2022-05-12 2023-11-16 Genmab A/S Binding agents capable of binding to cd27 in combination therapy
WO2023218046A1 (en) 2022-05-12 2023-11-16 Genmab A/S Binding agents capable of binding to cd27 in combination therapy
EP4285926A1 (en) 2022-05-30 2023-12-06 LAVA Therapeutics N.V. Combination treatment for chronic lymphocytic leukemia
US11845805B2 (en) 2020-09-10 2023-12-19 Genmab A/S Bispecific antibody against CD3 and CD20 in combination therapy for treating diffuse large B-cell lymphoma
EP4292609A1 (en) 2022-06-15 2023-12-20 LAVA Therapeutics N.V. Compositions comprising antibodies that bind gamma-delta t cell receptors
EP4292610A1 (en) 2022-06-15 2023-12-20 LAVA Therapeutics N.V. Variant antibodies that bind gamma-delta t cell receptors
US11858995B2 (en) 2020-09-10 2024-01-02 Genmab A/S Bispecific antibodies against CD3 and CD20 for treating chronic lymphocytic leukemia
US11866498B2 (en) 2018-02-08 2024-01-09 Genentech, Inc. Bispecific antigen-binding molecules and methods of use
WO2024010445A1 (en) 2022-07-06 2024-01-11 ACADEMISCH ZIEKENHUIS LEIDEN (h.o.d.n. LUMC) Bispecific antibody and uses thereof
US11926859B2 (en) 2012-04-20 2024-03-12 Merus N.V. Methods and means for the production of Ig-like molecules
WO2024079009A1 (en) 2022-10-10 2024-04-18 F. Hoffmann-La Roche Ag Combination therapy of a gprc5d tcb and proteasome inhibitors
WO2024079010A1 (en) 2022-10-10 2024-04-18 F. Hoffmann-La Roche Ag Combination therapy of a gprc5d tcb and cd38 antibodies
WO2024079015A1 (en) 2022-10-10 2024-04-18 F. Hoffmann-La Roche Ag Combination therapy of a gprc5d tcb and imids
WO2024094660A1 (en) 2022-10-31 2024-05-10 Genmab A/S Cd38 antibodies and uses thereof
US11981737B2 (en) 2016-11-18 2024-05-14 Hoffmann-La Roche Inc. Anti-HLA-G antibodies and use thereof
WO2024100170A1 (en) 2022-11-11 2024-05-16 F. Hoffmann-La Roche Ag Antibodies binding to hla-a*02/foxp3
WO2024104933A1 (en) 2022-11-15 2024-05-23 F. Hoffmann-La Roche Ag Antigen binding molecules
WO2024136658A1 (en) 2022-12-23 2024-06-27 Merus N.V. Combination treatment of cancers using an antibody that binds at least egfr and an immune checkpoint inhibitor
WO2024133330A1 (en) 2022-12-20 2024-06-27 Merus N.V. FAP BINDING DOMAINS AND BISPECIFIC BINDING MOIETIES THAT BIND FAP AND TGF-βRII
WO2024144401A1 (en) 2022-12-28 2024-07-04 Merus N.V. Treatment of cancer with a combination of an antibody that binds egfr and cytotoxic drugs
WO2024143442A1 (ja) 2022-12-27 2024-07-04 中外製薬株式会社 会合が制御されたポリペプチド
WO2024153722A1 (en) 2023-01-20 2024-07-25 F. Hoffmann-La Roche Ag Immunoconjugates
WO2024156672A1 (en) 2023-01-25 2024-08-02 F. Hoffmann-La Roche Ag Antibodies binding to csf1r and cd3
WO2024163494A1 (en) 2023-01-31 2024-08-08 F. Hoffmann-La Roche Ag Methods and compositions for treating non-small cell lung cancer and triple-negative breast cancer
WO2024163009A1 (en) 2023-01-31 2024-08-08 Genentech, Inc. Methods and compositions for treating urothelial bladder cancer
WO2024184287A1 (en) 2023-03-06 2024-09-12 F. Hoffmann-La Roche Ag Combination therapy of an anti-egfrviii/anti-cd3 antibody and an tumor-targeted 4-1bb agonist
WO2024188965A1 (en) 2023-03-13 2024-09-19 F. Hoffmann-La Roche Ag Combination therapy employing a pd1-lag3 bispecific antibody and an hla-g t cell bispecific antibody
US12103982B2 (en) 2014-11-20 2024-10-01 Hoffmann-La Roche Inc. T cell activating bispecific antigen binding molecules
EP4438624A1 (en) 2023-03-27 2024-10-02 LAVA Therapeutics N.V. Antibodies that bind nectin-4 and gamma-delta t cell receptors
WO2024200573A1 (en) 2023-03-27 2024-10-03 LAVA Therapeutics N.V. Nectin-4 binding agents and methods of use
WO2024208776A1 (en) 2023-04-03 2024-10-10 F. Hoffmann-La Roche Ag Agonistic split antibodies
WO2024208777A1 (en) 2023-04-03 2024-10-10 F. Hoffmann-La Roche Ag All-in-one agonistic antibodies
WO2024208898A1 (en) 2023-04-05 2024-10-10 Genmab A/S Pharmaceutical compositions comprising antibodies binding to cd30 and cd3
US12139553B2 (en) 2014-11-20 2024-11-12 Hoffmann-La Roche Inc. T cell activating bispecific antigen binding molecules
WO2024231320A1 (en) 2023-05-08 2024-11-14 F. Hoffmann-La Roche Ag Targeted interferon alpha fusion proteins and methods of use
EP4464333A2 (en) 2015-12-01 2024-11-20 Genmab B.V. Anti-death receptor antibodies and methods of use thereof
WO2024235862A1 (en) 2023-05-12 2024-11-21 Genmab A/S Antibodies capable of binding to ox40, variants thereof and uses thereof
WO2024238537A1 (en) 2023-05-16 2024-11-21 F. Hoffmann-La Roche Ag Pd-1 -regulated il-2 immunocytokine and uses thereof
US12157770B2 (en) 2018-04-18 2024-12-03 Hoffmann-La Roche Inc. Anti-HLA-G antibodies and methods of use thereof
WO2024263195A1 (en) 2023-06-23 2024-12-26 Genentech, Inc. Methods for treatment of liver cancer
WO2024263904A1 (en) 2023-06-23 2024-12-26 Genentech, Inc. Methods for treatment of liver cancer
WO2025003280A1 (en) 2023-06-30 2025-01-02 Genmab A/S Antibodies binding to fibroblast activation protein alpha and death receptor 4
US12195551B2 (en) 2017-05-17 2025-01-14 Merus N.V. Combination of an ErbB-2/ErbB-3 bispecific antibody with endocrine therapy for breast cancer
WO2025012118A2 (en) 2023-07-07 2025-01-16 LAVA Therapeutics N.V. 5t4 binding agents and methods of use
WO2025021838A1 (en) 2023-07-26 2025-01-30 F. Hoffmann-La Roche Ag Antibodies binding to cd3
WO2025024780A1 (en) 2023-07-27 2025-01-30 LAVA Therapeutics N.V. Antibodies that bind gamma-delta t cell receptors for the treatment of cancer
WO2025027511A1 (en) 2023-07-30 2025-02-06 Janssen Biotech, Inc. Molecules that bind to mutant calreticulin and uses thereof
WO2025042742A1 (en) 2023-08-18 2025-02-27 Bristol-Myers Squibb Company Compositions comprising antibodies that bind bcma and cd3 and methods of treatment
WO2025056180A1 (en) 2023-09-15 2025-03-20 BioNTech SE Methods of treatment using agents binding to epcam and cd137 in combination with pd-1 axis binding antagonists
WO2025105961A1 (en) 2023-11-17 2025-05-22 Merus N.V. COMBINATION THERAPY INCLUDING ANTIBODIES THAT BIND EGFR AND cMET.
WO2025114541A1 (en) 2023-11-30 2025-06-05 Genmab A/S Antibodies capable of binding to ox40 in combination therapy
WO2025125386A1 (en) 2023-12-14 2025-06-19 F. Hoffmann-La Roche Ag Antibodies that bind to folr1 and methods of use
WO2025181189A1 (en) 2024-03-01 2025-09-04 F. Hoffmann-La Roche Ag Antibodies binding to cd3
WO2025202147A1 (en) 2024-03-27 2025-10-02 F. Hoffmann-La Roche Ag Interleukin-7 immunoconjugates
US12435154B2 (en) 2020-05-08 2025-10-07 Genmab A/S Bispecific antibodies against CD3 and CD20
WO2025221138A1 (en) 2024-04-15 2025-10-23 Merus N.V. Cd3 binding domains
WO2025219504A1 (en) 2024-04-19 2025-10-23 F. Hoffmann-La Roche Ag Treatment of ophthalmologic diseases
WO2025230409A1 (en) 2024-04-30 2025-11-06 Merus N.V. Multispecific antibody-drug conjugates
WO2025230410A1 (en) 2024-04-30 2025-11-06 Merus N.V. Multispecific antibody-drug conjugates
WO2025233431A1 (en) 2024-05-07 2025-11-13 Immatics Biotechnologies Gmbh Heteromeric proteins comprising three heteromerization improving substitution, production, combinations and applications thereof

Families Citing this family (223)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SI1523496T1 (sl) † 2002-07-18 2011-11-30 Merus B V Rekombinantno proizvajanje zmesi protiteles
USRE47770E1 (en) 2002-07-18 2019-12-17 Merus N.V. Recombinant production of mixtures of antibodies
US20100069614A1 (en) 2008-06-27 2010-03-18 Merus B.V. Antibody producing non-human mammals
EP2395017A3 (en) 2003-05-30 2012-12-19 Merus B.V. Design and use of paired variable regions of specific binding molecules
DK3050963T3 (da) 2005-03-31 2019-12-09 Chugai Pharmaceutical Co Ltd Fremgangsmåde til fremstilling af polypeptid ved regulering af arrangement
DK2009101T3 (en) 2006-03-31 2018-01-15 Chugai Pharmaceutical Co Ltd Antibody modification method for purification of a bispecific antibody
EP2006381B1 (en) 2006-03-31 2016-02-03 Chugai Seiyaku Kabushiki Kaisha Method for controlling blood pharmacokinetics of antibodies
MY170607A (en) 2006-09-07 2019-08-20 Crucell Holland Bv Human binding molecules capable of neutralizing influenza virus h5n1 and uses thereof
CA2700701C (en) 2007-09-26 2020-12-29 Chugai Seiyaku Kabushiki Kaisha Method of modifying isoelectric point of antibody via amino acid substitution in cdr
CL2008002886A1 (es) 2007-09-26 2009-12-04 Chugai Pharmaceutical Co Ltd Region constante de un anticuerpo humano; anticuerpo anti-receptor de interleucina-6 (il-6) y composicion farmaceutica que la comprende.
WO2010107110A1 (ja) 2009-03-19 2010-09-23 中外製薬株式会社 抗体定常領域改変体
EP2826789A1 (en) 2009-03-19 2015-01-21 Chugai Seiyaku Kabushiki Kaisha Antibody constant region variant
US9493578B2 (en) 2009-09-02 2016-11-15 Xencor, Inc. Compositions and methods for simultaneous bivalent and monovalent co-engagement of antigens
EP2481752B1 (en) 2009-09-24 2016-11-09 Chugai Seiyaku Kabushiki Kaisha Modified antibody constant regions
US20130045492A1 (en) 2010-02-08 2013-02-21 Regeneron Pharmaceuticals, Inc. Methods For Making Fully Human Bispecific Antibodies Using A Common Light Chain
US9796788B2 (en) 2010-02-08 2017-10-24 Regeneron Pharmaceuticals, Inc. Mice expressing a limited immunoglobulin light chain repertoire
DK2505654T4 (da) 2010-02-08 2020-07-27 Regeneron Pharma Mus med fælles letkæde
WO2011108714A1 (ja) 2010-03-04 2011-09-09 中外製薬株式会社 抗体定常領域改変体
CN105585630B (zh) 2010-07-29 2020-09-15 Xencor公司 具有修改的等电点的抗体
MX355060B (es) 2010-11-17 2018-04-03 Chugai Pharmaceutical Co Ltd Molecula multiespecifica de union a antigeno que tiene funcion alternativa a la funcion del factor viii de coagulacion sanguinea.
MY172718A (en) 2011-08-05 2019-12-11 Regeneron Pharma Humanized universal light chain mice
CA2791109C (en) 2011-09-26 2021-02-16 Merus B.V. Generation of binding molecules
US12466897B2 (en) 2011-10-10 2025-11-11 Xencor, Inc. Heterodimeric human IgG1 polypeptides with isoelectric point modifications
US10851178B2 (en) 2011-10-10 2020-12-01 Xencor, Inc. Heterodimeric human IgG1 polypeptides with isoelectric point modifications
MX359775B (es) 2011-10-31 2018-10-10 Chugai Pharmaceutical Co Ltd Molecula enlazada al antigeno que tiene conjugacion regulada entre cadena ligera y cadena pesada.
WO2014006217A1 (en) 2012-07-06 2014-01-09 Genmab B.V. Dimeric protein with triple mutations
LT2900694T (lt) 2012-09-27 2018-11-12 Merus N.V. Bispecifiniai igg antikūnai kaip t ląstelių aktyvatoriai
KR101522954B1 (ko) 2012-11-27 2015-05-27 아주대학교산학협력단 항체 중쇄불변부위의 이종이중체 고효율 형성을 유도하는 ch3 도메인 변이체 쌍, 이의 제조방법, 및 용도
US11053316B2 (en) 2013-01-14 2021-07-06 Xencor, Inc. Optimized antibody variable regions
US9605084B2 (en) 2013-03-15 2017-03-28 Xencor, Inc. Heterodimeric proteins
US10968276B2 (en) 2013-03-12 2021-04-06 Xencor, Inc. Optimized anti-CD3 variable regions
US9701759B2 (en) 2013-01-14 2017-07-11 Xencor, Inc. Heterodimeric proteins
US10131710B2 (en) 2013-01-14 2018-11-20 Xencor, Inc. Optimized antibody variable regions
US10487155B2 (en) 2013-01-14 2019-11-26 Xencor, Inc. Heterodimeric proteins
DK2943511T3 (da) 2013-01-14 2019-10-21 Xencor Inc Nye heterodimeriske proteiner
WO2014113510A1 (en) 2013-01-15 2014-07-24 Xencor, Inc. Rapid clearance of antigen complexes using novel antibodies
US10106624B2 (en) 2013-03-15 2018-10-23 Xencor, Inc. Heterodimeric proteins
DK2970486T3 (en) 2013-03-15 2018-08-06 Xencor Inc MODULATION OF T-CELLS WITH BISPECIFIC ANTIBODIES AND FC-FUSIONS
US10519242B2 (en) 2013-03-15 2019-12-31 Xencor, Inc. Targeting regulatory T cells with heterodimeric proteins
US10858417B2 (en) 2013-03-15 2020-12-08 Xencor, Inc. Heterodimeric proteins
MX382731B (es) 2013-09-27 2025-03-13 Chugai Pharmaceutical Co Ltd Metodo para producir heteromultimeros de polipeptidos.
BR112016021572A2 (pt) 2014-03-21 2017-10-03 Regeneron Pharma Métodos para preparar um animal não humano geneticamente modificado e para produzir proteínas de ligação, hibridoma, ácido nucleico, célula, e, proteína de ligação
BR112016022385A2 (pt) 2014-03-28 2018-06-19 Xencor, Inc anticorpos específicos que se ligam a cd38 e cd3
MA40764A (fr) 2014-09-26 2017-08-01 Chugai Pharmaceutical Co Ltd Agent thérapeutique induisant une cytotoxicité
US10259887B2 (en) 2014-11-26 2019-04-16 Xencor, Inc. Heterodimeric antibodies that bind CD3 and tumor antigens
CN107406512A (zh) 2014-11-26 2017-11-28 Xencor公司 结合cd3和cd38的异二聚体抗体
KR102689285B1 (ko) 2014-11-26 2024-07-31 젠코어 인코포레이티드 Ｃｄ3 및 종양 항원과 결합하는 이종이량체 항체
WO2016105450A2 (en) 2014-12-22 2016-06-30 Xencor, Inc. Trispecific antibodies
CN105820251B (zh) * 2015-01-08 2019-10-15 江苏康宁杰瑞生物制药有限公司 具有共同轻链的双特异性抗体或抗体混合物
WO2016141387A1 (en) 2015-03-05 2016-09-09 Xencor, Inc. Modulation of t cells with bispecific antibodies and fc fusions
CA2979702A1 (en) 2015-03-19 2016-09-22 Regeneron Pharmaceuticals, Inc. Non-human animals that select for light chain variable regions that bind antigen
WO2016159213A1 (ja) 2015-04-01 2016-10-06 中外製薬株式会社 ポリペプチド異種多量体の製造方法
PL3115376T3 (pl) 2015-07-10 2019-01-31 Merus N.V. Ludzkie przeciwciało wiążące cd3
KR101851380B1 (ko) 2015-10-12 2018-04-23 아주대학교산학협력단 효모접합을 이용한 항체 ch3 도메인 이종이중체 돌연변이쌍 제조 방법 및 이에 의하여 제조된 ch3 돌연변이체 쌍
AU2016344665C1 (en) 2015-10-29 2023-07-27 F. Hoffmann-La Roche Ag Anti-variant Fc-region antibodies and methods of use
AU2016365742A1 (en) 2015-12-07 2018-06-21 Xencor, Inc. Heterodimeric antibodies that bind CD3 and PSMA
CN106883297B (zh) * 2015-12-16 2019-12-13 苏州康宁杰瑞生物科技有限公司 基于ch3结构域的异二聚体分子、其制备方法及用途
MX2018007781A (es) 2015-12-28 2018-09-05 Chugai Pharmaceutical Co Ltd Metodo para promover la eficiencia de purificacion del polipeptido que contiene la region de fragmento cristalizable (fc).
WO2017159287A1 (ja) 2016-03-14 2017-09-21 中外製薬株式会社 癌の治療に用いるための細胞傷害誘導治療剤
US11291721B2 (en) 2016-03-21 2022-04-05 Marengo Therapeutics, Inc. Multispecific and multifunctional molecules and uses thereof
EP3449940B1 (en) 2016-04-28 2025-11-19 Chugai Seiyaku Kabushiki Kaisha Antibody-containing preparation
US10787518B2 (en) 2016-06-14 2020-09-29 Xencor, Inc. Bispecific checkpoint inhibitor antibodies
KR20190020341A (ko) 2016-06-28 2019-02-28 젠코어 인코포레이티드 소마토스타틴 수용체 2에 결합하는 이종이량체 항체
DK3511340T3 (da) 2016-08-10 2025-01-02 Univ Ajou Ind Academic Coop Found Heterodimerisk fc-fusioneret cytokin og farmaceutisk sammensætning omfattende det samme
US10793632B2 (en) 2016-08-30 2020-10-06 Xencor, Inc. Bispecific immunomodulatory antibodies that bind costimulatory and checkpoint receptors
JP6785372B2 (ja) 2016-09-30 2020-11-18 エフ．ホフマン−ラ ロシュ アーゲーＦ． Ｈｏｆｆｍａｎｎ−Ｌａ Ｒｏｃｈｅ Ａｋｔｉｅｎｇｅｓｅｌｌｓｃｈａｆｔ 多重特異性分子の機能分析のためのｓｐｒに基づく二重結合アッセイ
SG11201903302UA (en) 2016-10-14 2019-05-30 Xencor Inc Bispecific heterodimeric fusion proteins containing il-15/il-15ralpha fc-fusion proteins and pd-1 antibody fragments
WO2018129522A1 (en) 2017-01-09 2018-07-12 Torch Therapeutics Conditionally effective bispecific therapeutics
JP2020506971A (ja) 2017-02-08 2020-03-05 ドラゴンフライ セラピューティクス， インコーポレイテッド ナチュラルキラー細胞の活性化のための多重特異性結合タンパク質およびがんを処置するためのその治療的使用
US20200291089A1 (en) 2017-02-16 2020-09-17 Elstar Therapeutics, Inc. Multifunctional molecules comprising a trimeric ligand and uses thereof
SG11201907646YA (en) 2017-02-20 2019-09-27 Dragonfly Therapeutics Inc Proteins binding her2, nkg2d and cd16
US12054544B2 (en) * 2017-02-24 2024-08-06 Chugai Seiyaku Kabushiki Kaisha Compositions comprising antigen-binding molecules
MA48723A (fr) 2017-03-10 2020-04-08 Hoffmann La Roche Procédé de production d'anticorps multispécifiques
CA3058341A1 (en) 2017-03-31 2018-10-04 Merus N.V. Antibodies for the treatment of erbb-2/erbb-3 positive tumors
CN111032081A (zh) 2017-03-31 2020-04-17 美勒斯公司 ErbB-2和ErbB-3靶向剂和双特异性抗体
KR102629905B1 (ko) * 2017-04-01 2024-01-30 베이징 한미 파마슈티컬 컴퍼니 리미티드 항-pd-l1/항-pd-1 천연 항체 구조-유사 헤테로다이머 이중특이성 항체 및 그의 제조
JOP20190222A1 (ar) 2017-04-11 2019-09-24 Zymeworks Inc الأجسام المضادة ثنائية النوعية المضادة لـ pd-l1 والمضادة لـ tim-3
WO2018222901A1 (en) 2017-05-31 2018-12-06 Elstar Therapeutics, Inc. Multispecific molecules that bind to myeloproliferative leukemia (mpl) protein and uses thereof
GB201709970D0 (en) 2017-06-22 2017-08-09 Kymab Ltd Bispecific antigen-binding molecules
US11084863B2 (en) 2017-06-30 2021-08-10 Xencor, Inc. Targeted heterodimeric Fc fusion proteins containing IL-15 IL-15alpha and antigen binding domains
AR112603A1 (es) 2017-07-10 2019-11-20 Lilly Co Eli Anticuerpos biespecíficos inhibidores de punto de control
WO2019014912A1 (zh) * 2017-07-21 2019-01-24 赵磊 一种异二聚体蛋白及其制备方法
CA3076027A1 (en) 2017-11-01 2019-05-09 F. Hoffmann-La Roche Ag Bispecific 2+1 contorsbodies
BR112020008393A2 (pt) 2017-11-01 2020-11-03 Chugai Seiyaku Kabushiki Kaisha variante e isoforma de anticorpos com atividade biológica reduzida
JP2021500902A (ja) 2017-11-01 2021-01-14 エフ・ホフマン−ラ・ロシュ・アクチェンゲゼルシャフト 新規ｔｎｆファミリーリガンド三量体含有抗原結合分子
US10981992B2 (en) 2017-11-08 2021-04-20 Xencor, Inc. Bispecific immunomodulatory antibodies that bind costimulatory and checkpoint receptors
AU2018366199A1 (en) 2017-11-08 2020-05-28 Xencor, Inc. Bispecific and monospecific antibodies using novel anti-PD-1 sequences
EP3717008A1 (en) 2017-12-01 2020-10-07 Merus N.V. Use of bispecific antibody and il-15 for combination therapy
BR112020011295A2 (pt) * 2017-12-04 2020-11-24 Beijing Hanmi Pharmaceutical Co., Ltd. anticorpo biespecífico anti-pd-l1/anti-cd47 com estrutura semelhante ao anticorpo natural e em forma de heterodímero e preparação do mesmo
CA3086199A1 (en) 2017-12-19 2019-06-27 Xencor, Inc. Engineered il-2 fc fusion proteins
WO2019122054A1 (en) 2017-12-22 2019-06-27 F. Hoffmann-La Roche Ag Depletion of light chain mispaired antibody variants by hydrophobic interaction chromatography
WO2019139987A1 (en) 2018-01-09 2019-07-18 Elstar Therapeutics, Inc. Calreticulin binding constructs and engineered t cells for the treatment of diseases
WO2019148410A1 (en) 2018-02-01 2019-08-08 Merck Sharp & Dohme Corp. Anti-pd-1 antibodies
WO2019148412A1 (en) 2018-02-01 2019-08-08 Merck Sharp & Dohme Corp. Anti-pd-1/lag3 bispecific antibodies
PE20210375A1 (es) 2018-02-08 2021-03-02 Dragonfly Therapeutics Inc Terapia de combinacion del cancer que incluye proteinas de union multiespecificas que activan a las celulas asesinas naturales
FI3749346T3 (fi) 2018-02-08 2024-09-04 Dragonfly Therapeutics Inc Nkg2d-reseptoriin kohdistuvat vasta-aineen vaihtelevan domeenin yhdistelmät
JP2021512624A (ja) * 2018-02-11 2021-05-20 ベイジン・ハンミ・ファーマシューティカル・カンパニー・リミテッドＢｅｉｊｉｎｇ Ｈａｎｍｉ Ｐｈａｒｍａｃｅｕｔｉｃａｌ Ｃｏ．， Ｌｔｄ． 抗ｐｄ−１／抗ｖｅｇｆ天然抗体構造様ヘテロ二量体型二重特異性抗体及びその製造
MX2020008684A (es) 2018-02-20 2020-12-07 Dragonfly Therapeutics Inc Proteinas de union multiespecificas que se unen a cd33, nkg2d y cd16 y metodos de uso.
US20210009711A1 (en) 2018-03-14 2021-01-14 Elstar Therapeutics, Inc. Multifunctional molecules and uses thereof
US12152073B2 (en) 2018-03-14 2024-11-26 Marengo Therapeutics, Inc. Multifunctional molecules that bind to calreticulin and uses thereof
AU2019247415A1 (en) 2018-04-04 2020-10-22 Xencor, Inc. Heterodimeric antibodies that bind fibroblast activation protein
EP3781204A4 (en) * 2018-04-17 2022-03-30 Invenra, Inc. BINDING MOLECULES
JP2021520829A (ja) 2018-04-18 2021-08-26 ゼンコア インコーポレイテッド ＩＬ−１５／ＩＬ−１５ＲＡ Ｆｃ融合タンパク質およびＴＩＭ−３抗原結合ドメインを含む、ＴＩＭ−３標的化ヘテロ二量体融合タンパク質
CN112867734A (zh) 2018-04-18 2021-05-28 Xencor股份有限公司 包含IL-15/IL-15Ra Fc融合蛋白和PD-1抗原结合结构域的靶向PD-1的异源二聚体融合蛋白及其用途
MX2020012286A (es) 2018-05-16 2021-04-28 Janssen Biotech Inc Metodos para tratar canceres y potenciar la eficacia de agentes terapeuticos para el redireccionamiento de celulas t.
CN112955465A (zh) 2018-07-03 2021-06-11 马伦戈治疗公司 抗tcr抗体分子及其用途
US12384851B2 (en) 2018-08-08 2025-08-12 Dragonfly Therapeutics, Inc. Multi-specific binding proteins that bind BCMA, NKG2D and CD16, and methods of use
JP7482363B2 (ja) 2018-08-08 2024-05-14 ドラゴンフライ セラピューティクス， インコーポレイテッド Ｎｋｇ２ｄ、ｃｄ１６及び腫瘍関連抗原に結合するタンパク質
EA202091888A1 (ru) 2018-08-08 2020-10-23 Драгонфлай Терапьютикс, Инк. Вариабельные домены антител, нацеленные на рецептор nkg2d
GB2576914A (en) 2018-09-06 2020-03-11 Kymab Ltd Antigen-binding molecules comprising unpaired variable domains produced in mammals
KR102884523B1 (ko) 2018-10-03 2025-11-10 젠코어 인코포레이티드 IL-12 이종이량체 Fc-융합 단백질
TWI891611B (zh) 2018-10-23 2025-08-01 美商蜻蜓醫療公司 異二聚體fc融合蛋白
US20220041719A1 (en) 2018-12-05 2022-02-10 Morphosys Ag Multispecific antigen-binding molecules
TWI874341B (zh) 2018-12-18 2025-03-01 美商健生生物科技公司 產生異二聚體抗體之方法
GB201820687D0 (en) 2018-12-19 2019-01-30 Kymab Ltd Antagonists
WO2020130829A1 (en) 2018-12-20 2020-06-25 Merus N.V. Clec12axcd3 bispecific antibodies and methods for the treatment of disease
SI3723858T1 (sl) 2018-12-21 2022-04-29 Kymab Limited Bispecifično protitelo FIXAXFX s skupno lahko verigo
TWI852977B (zh) 2019-01-10 2024-08-21 美商健生生物科技公司 前列腺新抗原及其用途
BR112021016092A2 (pt) * 2019-02-14 2021-10-26 Merus N.V. Produção de composições que compreendem dois ou mais anticorpos
CN114409798A (zh) * 2019-02-14 2022-04-29 美勒斯公司 制备包含两种或更多种抗体的组合物
SG11202108737RA (en) 2019-02-14 2021-09-29 Merus Nv Combinations of binding moieties that bind egfr, her2 and her3.
GB2599227B (en) 2019-02-21 2024-05-01 Marengo Therapeutics Inc Multifunctional molecules that bind to T cells and uses thereof to treat autoimmune disorders
JP7710373B2 (ja) 2019-02-21 2025-07-18 マレンゴ・セラピューティクス，インコーポレーテッド Ｔ細胞関連のがん細胞に結合する多機能性分子およびその使用
JP2022521751A (ja) 2019-02-21 2022-04-12 マレンゴ・セラピューティクス，インコーポレーテッド 抗ｔｃｒ抗体分子およびその使用
WO2020172605A1 (en) 2019-02-21 2020-08-27 Elstar Therapeutics, Inc. Antibody molecules that bind to nkp30 and uses thereof
SG11202109056TA (en) 2019-02-21 2021-09-29 Marengo Therapeutics Inc Multifunctional molecules that bind to calreticulin and uses thereof
SG11202109406TA (en) 2019-03-01 2021-09-29 Xencor Inc Heterodimeric antibodies that bind enpp3 and cd3
WO2020198683A1 (en) * 2019-03-28 2020-10-01 Ab Studio Inc. Heteromultimeric proteins and methods of use thereof
CN113677701B (zh) 2019-03-29 2025-02-07 豪夫迈·罗氏有限公司 产生亲合结合多特异性抗体的方法
JP7249432B2 (ja) 2019-03-29 2023-03-30 エフ． ホフマン－ラ ロシュ アーゲー 多価分子の機能分析のための、ｓｐｒをベースとする結合アッセイ
BR112021021210A2 (pt) 2019-04-25 2021-12-21 Hoffmann La Roche Conjunto de polipeptídeos, método para gerar um polipeptídeo heterodimérico, primeiro polipeptídeo precursor heterodimérico, segundo polipeptídeo precursor heterodimérico e composição farmacêutica
CA3132275A1 (en) * 2019-04-25 2020-10-29 Ulrich Brinkmann Generation of antibody-derived polypeptides by polypeptide chain exchange
KR20220004052A (ko) 2019-04-25 2022-01-11 에프. 호프만-라 로슈 아게 폴리펩티드 사슬 교환에 의해 활성화된 치료용 다중특이적 폴리펩티드
NZ782091A (en) * 2019-05-09 2025-08-29 Merus Nv Variant domains for multimerizing proteins and separation thereof
CN119470884A (zh) 2019-05-13 2025-02-18 豪夫迈·罗氏有限公司 抑制干扰的药代动力学免疫测定
JP7317353B2 (ja) * 2019-06-18 2023-07-31 株式会社プロテイン・エクスプレス ブレビバチルス菌によるヘテロ二量体タンパク質の生産
WO2020254357A1 (en) 2019-06-19 2020-12-24 F. Hoffmann-La Roche Ag Method for the generation of a protein expressing cell by targeted integration using cre mrna
JP7450647B2 (ja) 2019-06-26 2024-03-15 エフ． ホフマン－ラ ロシュ アーゲー Ｓｉｒｔ－１遺伝子ノックアウトを有する哺乳類細胞株
NZ785553A (en) 2019-07-26 2025-07-25 Janssen Biotech Inc Proteins comprising kallikrein related peptidase 2 antigen binding domains and their uses
JOP20220035A1 (ar) 2019-08-15 2023-01-30 Janssen Biotech Inc مواد وطرق لتحسين أجزاء المتغير أحادي السلسلة
TWI873210B (zh) 2019-10-24 2025-02-21 荷蘭商美勒斯公司 用於治療患有ｈｅｒ２及ｈｅｒ３陽性癌症之對象的手段及方法
JP7737983B2 (ja) 2019-11-01 2025-09-11 インノコア テクノロジーズ ビー．ブイ． 抗体又は巨大タンパク質の延長された放出の為の剤形物
KR20220110233A (ko) 2019-12-02 2022-08-05 리제너론 파마슈티칼스 인코포레이티드 펩티드-mhc ii 단백질 작제물 및 그의 용도
JP7692916B2 (ja) 2020-01-02 2025-06-16 エフ． ホフマン－ラ ロシュ アーゲー 脳内の治療用抗体の量を決定するための方法
WO2021138407A2 (en) 2020-01-03 2021-07-08 Marengo Therapeutics, Inc. Multifunctional molecules that bind to cd33 and uses thereof
CA3166629A1 (en) 2020-01-03 2021-07-08 Marengo Therapeutics, Inc. Anti-tcr antibody molecules and uses thereof
KR20220144377A (ko) 2020-01-30 2022-10-26 우모자 바이오파마 인코포레이티드 이중특이적 형질도입 촉진제
TW202144388A (zh) 2020-02-14 2021-12-01 美商健生生物科技公司 在卵巢癌中表現之新抗原及其用途
TW202144389A (zh) 2020-02-14 2021-12-01 美商健生生物科技公司 在多發性骨髓瘤中表現之新抗原及其用途
TW202144418A (zh) 2020-03-13 2021-12-01 美商健生生物科技公司 唾液酸結合Ｉｇ樣凝集素結合的材料及方法
WO2021190980A1 (en) 2020-03-22 2021-09-30 Quadrucept Bio Limited Multimers for viral strain evolution
WO2021203103A2 (en) * 2020-04-03 2021-10-07 Somasekar Seshagiri Ace2 receptor polymorphisms and varying susceptibility to sars-cov-2, methods for diagnosis and treatment
KR20230028242A (ko) 2020-04-24 2023-02-28 마렝고 테라퓨틱스, 인크. T 세포 관련 암 세포에 결합하는 다중기능성 분자 및 그것의 용도
TW202208428A (zh) 2020-05-06 2022-03-01 美商蜻蜓醫療公司 結合ｎｋｇ２ｄ、ｃｄ１６及ｃｌｅｃ１２ａ之蛋白質
KR20230010240A (ko) 2020-05-11 2023-01-18 에프. 호프만-라 로슈 아게 변형된 pbmc 및 면역접합체를 이용한 병용 요법
US11919956B2 (en) 2020-05-14 2024-03-05 Xencor, Inc. Heterodimeric antibodies that bind prostate specific membrane antigen (PSMA) and CD3
KR20230017841A (ko) 2020-05-27 2023-02-06 얀센 바이오테크 인코포레이티드 Cd3 항원 결합 도메인을 포함하는 단백질 및 이의 용도
EP4173696A4 (en) 2020-06-24 2023-12-20 Asahi Kasei Medical Co., Ltd. METHOD FOR EVALUATING A SOLUTION CONTAINING PROTEINS
WO2022009052A2 (en) 2020-07-06 2022-01-13 Janssen Biotech, Inc. Prostate neoantigens and their uses
JP2023536818A (ja) 2020-07-29 2023-08-30 ヤンセン バイオテツク，インコーポレーテツド Ｈｌａ－ｇ抗原結合ドメインを含むタンパク質及びそれらの使用
KR20230166150A (ko) 2020-08-19 2023-12-06 젠코어 인코포레이티드 항-cd28 조성물
JP2023542080A (ja) 2020-08-26 2023-10-05 マレンゴ・セラピューティクス，インコーポレーテッド カルレティキュリンに結合する多機能性分子およびその使用
EP4204458A4 (en) 2020-08-26 2024-10-09 Marengo Therapeutics, Inc. METHODS FOR DETECTION OF TRBC1 OR TRBC2
AU2021331076A1 (en) 2020-08-26 2023-04-06 Marengo Therapeutics, Inc. Antibody molecules that bind to NKp30 and uses thereof
MX2023003328A (es) 2020-09-24 2023-03-27 Hoffmann La Roche Lineas de celulas de mamifero con desactivacion genica.
TW202231292A (zh) 2020-10-13 2022-08-16 美商健生生物科技公司 用於調節分化簇ｉｖ及／或ｖｉｉｉ的經生物工程改造之ｔ細胞介導之免疫力、材料、及其他方法
KR20230110523A (ko) 2020-10-22 2023-07-24 얀센 바이오테크 인코포레이티드 델타-유사 리간드 3(dll3) 항원 결합 영역을 포함하는 단백질 및 그의 용도
JP2023548375A (ja) 2020-11-04 2023-11-16 メルス ナムローゼ フェンノートシャップ Ｅｒｂｂ３変異陽性がんを有する対象を治療するための手段及び方法
KR20230107305A (ko) 2020-11-10 2023-07-14 상하이 치루 파마슈티컬 리서치 앤 디벨롭먼트 센터 리미티드 클라우딘 18a2 및 cd3에 대한 이중특이성 항체 및 이중특이성 항체의 용도
MX2023007309A (es) 2020-12-18 2023-08-14 Merus Nv Composicion de anticuerpos.
CN116601175A (zh) 2020-12-18 2023-08-15 豪夫迈·罗氏有限公司 用于靶向疗法的前体蛋白和试剂盒
JP2024501662A (ja) 2020-12-22 2024-01-15 エフ． ホフマン－ラ ロシュ アーゲー Ｘｂｐ１を標的とするオリゴヌクレオチド
CN117440969A (zh) * 2021-01-11 2024-01-23 阿迪马布有限责任公司 被工程化为优先进行ch3异源二聚化的变体ch3结构域、包括所述变体ch3结构域的多特异性抗体和其制备方法
EP4295154A1 (en) 2021-02-18 2023-12-27 F. Hoffmann-La Roche AG Method for resolving complex, multistep antibody interactions
WO2022184659A1 (en) 2021-03-01 2022-09-09 Quadrucept Bio Limited Antibody domains & multimers
EP4301774A4 (en) 2021-03-03 2025-08-13 Dragonfly Therapeutics Inc METHODS OF TREATING CANCER USING MULTI-SPECIFIC BINDING PROTEINS THAT BIND TO NKG2D, CD16, AND A TUMOR-ASSOCIATED ANTIGEN
WO2022192403A1 (en) 2021-03-09 2022-09-15 Xencor, Inc. Heterodimeric antibodies that bind cd3 and cldn6
JP2024509274A (ja) 2021-03-10 2024-02-29 ゼンコア インコーポレイテッド Ｃｄ３及びｇｐｃ３に結合するヘテロ二量体抗体
AU2022244453A1 (en) 2021-03-24 2023-11-09 Janssen Biotech, Inc. Antibody targeting cd22 and cd79b
CN117377694A (zh) 2021-03-24 2024-01-09 詹森生物科技公司 包含cd3抗原结合结构域的蛋白质及其用途
CN116897159A (zh) 2021-03-31 2023-10-17 江苏恒瑞医药股份有限公司 截短的taci多肽及其融合蛋白和用途
US20250099579A1 (en) 2021-04-08 2025-03-27 Marengo Therapeutics, Inc. Multispecific molecules binding to tcr and uses thereof
WO2022241398A1 (en) * 2021-05-10 2022-11-17 Medimmune, Llc Modified fcrn binding fragments with improved half-life
JP2024518081A (ja) 2021-05-12 2024-04-24 江▲蘇▼恒瑞医▲薬▼股▲フン▼有限公司 Ｒａｎｋｌとｎｇｆに特異的に結合する抗原結合分子およびその医薬用途
JP2024517907A (ja) 2021-05-14 2024-04-23 江蘇恒瑞医薬股▲ふん▼有限公司 抗原結合分子
EP4372001A4 (en) 2021-07-14 2025-02-12 Jiangsu Hengrui Pharmaceuticals Co., Ltd. ANTIGEN-BINDING MOLECULE SPECIFICALLY BINDING TO HGFR AND EGFR, AND PHARMACEUTICAL USE THEREOF
JP2024527977A (ja) 2021-07-27 2024-07-26 ノヴァブ， インコーポレイテッド 免疫エフェクター機能を有する操作されたｖｌｒｂ抗体
EP4377347A1 (en) 2021-07-27 2024-06-05 MorphoSys AG Combinations of antigen binding molecules
KR102351147B1 (ko) * 2021-08-02 2022-01-14 주식회사 바이오뱅크힐링 블라우티아 오베움 균주, 및 그의 유래의 소포체 및 그의 항염증 및 항균 용도
EP4405398A4 (en) * 2021-09-21 2025-10-29 Qilu Puget Sound Biotherapeutics Corp FC HETERODIMERS ALLOWING THE MANUFACTURE OF FUSION PROTEINS AND BI-SPECIFIC ANTIBODIES
CA3232472A1 (en) 2021-09-23 2023-03-30 Jiangsu Hengrui Pharmaceuticals Co., Ltd. Anti-klb antibodies and uses
US20250019455A1 (en) 2021-09-24 2025-01-16 Pharmaceutica Nv Proteins comprising cd20 binding domains, and uses thereof
WO2023051798A1 (zh) 2021-09-30 2023-04-06 江苏恒瑞医药股份有限公司 抗il23抗体融合蛋白及用途
CA3237357A1 (en) 2021-11-03 2023-05-11 Janssen Biotech, Inc. Methods of treating cancers and enhancing efficacy of bcmaxcd3 bispecific antibodies
MX2024006205A (es) 2021-11-22 2024-08-19 Janssen Biotech Inc Composiciones que comprenden agentes de unión multiespecíficos potenciados para una respuesta inmunitaria.
US20250043022A1 (en) 2022-02-07 2025-02-06 Jiangsu Hengrui Pharmaceuticals Co., Ltd. Antigen-binding molecule specifically binding to psma and cd3, and pharmaceutical use thereof
WO2023151661A1 (zh) 2022-02-11 2023-08-17 江苏恒瑞医药股份有限公司 免疫缀合物及其用途
CN118660910A (zh) 2022-03-14 2024-09-17 江苏恒瑞医药股份有限公司 特异性结合gprc5d和cd3的抗原结合分子及其医药用途
WO2023202967A1 (en) 2022-04-19 2023-10-26 F. Hoffmann-La Roche Ag Improved production cells
JP2025517572A (ja) 2022-06-03 2025-06-05 エフ． ホフマン－ラ ロシュ アーゲー 改良された産生細胞
CN119256017A (zh) 2022-06-23 2025-01-03 江苏恒瑞医药股份有限公司 特异性结合dll3和cd3的抗原结合分子及其医药用途
CN119546645A (zh) 2022-08-05 2025-02-28 江苏恒瑞医药股份有限公司 特异性结合gucy2c和cd3的抗原结合分子及其医药用途
CN120887995A (zh) 2022-08-05 2025-11-04 美勒斯公司 治疗去势抗性前列腺癌的手段和方法
JP2025535744A (ja) 2022-10-12 2025-10-28 エフ． ホフマン－ラ ロシュ アーゲー 細胞を分類するための方法
EP4613767A1 (en) 2022-11-01 2025-09-10 Shanghai Qilu Pharmaceutical Research and Development Centre Ltd. Bispecific antibody for glypican-3 and use thereof
IL320562A (en) 2022-11-02 2025-07-01 Janssen Biotech Inc Cancer treatment methods
JP2025537782A (ja) 2022-11-15 2025-11-20 エフ． ホフマン－ラ ロシュ アーゲー 活性化可能なエフェクタードメインを有する組換え結合タンパク質
JP2025538642A (ja) 2022-11-29 2025-11-28 江蘇恒瑞医薬股▲ふん▼有限公司 Ｃｌｄｎ１８．２／４－１ｂｂ結合タンパク質及びその医薬的使用
WO2024175031A1 (zh) 2023-02-21 2024-08-29 江苏恒瑞医药股份有限公司 Il-36r结合蛋白及其医药用途
WO2024213040A1 (zh) 2023-04-11 2024-10-17 浙江博锐生物制药有限公司 抗ror1抗体及其药物偶联物
WO2025007037A1 (en) 2023-06-28 2025-01-02 Dragonfly Therapeutics, Inc. Variant il-2 proteins
US20250066510A1 (en) 2023-08-07 2025-02-27 Janssen Biotech, Inc. Gucy2c antibodies and uses thereof
WO2025032069A1 (en) 2023-08-09 2025-02-13 F. Hoffmann-La Roche Ag Mono and multispecific anti-trem2 antibodies, methods and uses thereof
WO2025032071A1 (en) 2023-08-09 2025-02-13 F. Hoffmann-La Roche Ag Mono and multispecific anti-trem2 antibodies, methods and uses thereof
WO2025036892A1 (en) 2023-08-14 2025-02-20 Morphosys Ag Cycat halfbody molecules comprising sterically occluding moieties
WO2025059162A1 (en) 2023-09-11 2025-03-20 Dana-Farber Cancer Institute, Inc. Car-engager containing il-2 variants to enhance the functionality of car t cells
WO2025082777A1 (en) 2023-10-17 2025-04-24 Morphosys Ag Dual-targeting of muc16 and mesothelin co-expressing tumor cells by functional complementation of cycat® halfbody molecules
WO2025137454A1 (en) * 2023-12-21 2025-06-26 Amgen Inc. Stabilizing homodimer mutations for two cell heterodimer production
WO2025188180A1 (en) 2024-03-04 2025-09-12 Merus N.V. Erbb-2 and erbb3 targeting agents for use in the treatment of malignant cells or cancers that have elevated nrg1 expression
WO2025231408A2 (en) 2024-05-03 2025-11-06 Janssen Biotech, Inc. Methods for treating multiple myeloma with car-t cells and bispecific antibodies
WO2025228542A1 (en) 2024-05-03 2025-11-06 Genmab B.V. Method for co-expression of a plurality of antibodies
WO2025237931A1 (en) 2024-05-15 2025-11-20 F. Hoffmann-La Roche Ag Recombinant binding proteins with conditionally activatable t cell and nk cell recruiting effector domains
CN121135878A (zh) 2024-06-14 2025-12-16 康诺亚生物医药科技（成都）有限公司 针对ox40l和il-13的双特异性抗体及其用途

Citations (23)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0120694A2 (en) 1983-03-25 1984-10-03 Celltech Limited Processes for the production of multichain polypeptides or proteins
EP0187045A2 (en) 1984-12-28 1986-07-09 Elf Atochem S.A. Radiation curable macromonomer printing inks
US4801687A (en) 1986-10-27 1989-01-31 Bioprobe International, Inc. Monoclonal antibody purification process using protein A
US4816567A (en) 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
EP0314161A1 (en) 1987-10-28 1989-05-03 Bristol-Myers Squibb Company Human immunoglobulines produced by recombinant DNA techniques
EP0481790A2 (en) 1990-10-17 1992-04-22 The Wellcome Foundation Limited Antibody production
US5151504A (en) 1989-11-17 1992-09-29 E. R. Squibb & Sons, Inc. Method for purification of monoclonal antibodies
EP0523949A1 (en) 1991-07-15 1993-01-20 The Wellcome Foundation Limited Production of antibodies
WO1998050431A2 (en) 1997-05-02 1998-11-12 Genentech, Inc. A method for making multispecific antibodies having heteromultimeric and common components
WO2000063403A2 (en) 1999-04-15 2000-10-26 Crucell Holland B.V. Recombinant protein production in a human cell using sequences encoding adenovirus e1 protein
WO2003004704A2 (en) 2001-07-04 2003-01-16 Chromagenics B.V. Method of selecting dna sequence with transcription modulating activity using a vector comprising an element with a gene transcription repressing activity
WO2004009618A2 (en) 2002-07-18 2004-01-29 Crucell Holland B.V. Recombinant production of mixtures of antibodies
WO2004061104A2 (en) 2003-01-07 2004-07-22 Symphogen A/S Method for manufacturing recombinant polyclonal proteins
WO2007110205A2 (en) 2006-03-24 2007-10-04 Merck Patent Gmbh Engineered heterodimeric protein domains
WO2007147901A1 (en) 2006-06-22 2007-12-27 Novo Nordisk A/S Production of bispecific antibodies
WO2009080252A1 (en) 2007-12-21 2009-07-02 F. Hoffmann-La Roche Ag Bivalent, bispecific antibodies
WO2009080253A1 (en) 2007-12-21 2009-07-02 F. Hoffmann-La Roche Ag Bivalent, bispecific antibodies
WO2009080251A1 (en) 2007-12-21 2009-07-02 F. Hoffmann-La Roche Ag Bivalent, bispecific antibodies
WO2009089004A1 (en) 2008-01-07 2009-07-16 Amgen Inc. Method for making antibody fc-heterodimeric molecules using electrostatic steering effects
US20100015133A1 (en) 2005-03-31 2010-01-21 Chugai Seiyaku Kabushiki Kaisha Methods for Producing Polypeptides by Regulating Polypeptide Association
WO2010084197A1 (en) * 2009-01-26 2010-07-29 Genmab A/S Methods for producing mixtures of antibodies
WO2010129304A2 (en) 2009-04-27 2010-11-11 Oncomed Pharmaceuticals, Inc. Method for making heteromultimeric molecules
US20110195454A1 (en) * 2010-02-08 2011-08-11 Regeneron Pharmaceuticals, Inc. Common Light Chain Mouse

Family Cites Families (185)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4399216A (en) 1980-02-25 1983-08-16 The Trustees Of Columbia University Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials
US4634665A (en) 1980-02-25 1987-01-06 The Trustees Of Columbia University In The City Of New York Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials
US5179017A (en) 1980-02-25 1993-01-12 The Trustees Of Columbia University In The City Of New York Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials
US4599311A (en) 1982-08-13 1986-07-08 Kawasaki Glenn H Glycolytic promotersfor regulated protein expression: protease inhibitor
AU600885B2 (en) 1984-05-25 1990-08-30 Zymogenetics Inc. Stable DNA constructs
US5168062A (en) 1985-01-30 1992-12-01 University Of Iowa Research Foundation Transfer vectors and microorganisms containing human cytomegalovirus immediate-early promoter-regulatory DNA sequence
US4868103A (en) 1986-02-19 1989-09-19 Enzo Biochem, Inc. Analyte detection by means of energy transfer
ATE87659T1 (de) 1986-09-02 1993-04-15 Enzon Lab Inc Bindungsmolekuele mit einzelpolypeptidkette.
DE307667T1 (de) 1987-08-24 1991-09-05 Wacom Co., Ltd., Saitama Koordinateneingabesystem mit einem eingabeschreibstift.
US4937190A (en) 1987-10-15 1990-06-26 Wisconsin Alumni Research Foundation Translation enhancer
ES2035317T5 (es) 1987-11-09 1998-03-16 Becton Dickinson Co Metodo para analizar celulas hematopoyeticas en una muestra.
WO1989006692A1 (en) 1988-01-12 1989-07-27 Genentech, Inc. Method of treating tumor cells by inhibiting growth factor receptor function
US5223409A (en) 1988-09-02 1993-06-29 Protein Engineering Corp. Directed evolution of novel binding proteins
AU4308689A (en) 1988-09-02 1990-04-02 Protein Engineering Corporation Generation and selection of recombinant varied binding proteins
GB8909218D0 (en) 1989-04-22 1989-06-07 Medical Res Council Improvements in or relating to enhancers
DE69005908T2 (de) 1989-06-08 1994-04-28 Wistar Inst Philadelphia Monoklonale Antikörper zur Behandlung nach einem Kontakt mit Tollwutvirus.
WO1991000906A1 (en) 1989-07-12 1991-01-24 Genetics Institute, Inc. Chimeric and transgenic animals capable of producing human antibodies
US5030002A (en) 1989-08-11 1991-07-09 Becton, Dickinson And Company Method and apparatus for sorting particles with a moving catcher tube
US5959177A (en) 1989-10-27 1999-09-28 The Scripps Research Institute Transgenic plants expressing assembled secretory antibodies
DE69027683T2 (de) 1989-12-01 1997-02-13 Pharming Bv Herstellung rekombinanter polypeptide durch rinder und transgene methoden
ATE356869T1 (de) 1990-01-12 2007-04-15 Amgen Fremont Inc Bildung von xenogenen antikörpern
DE4006630A1 (de) 1990-03-03 1991-09-12 Behringwerke Ag Humane monoklonale antikoerper gegen tollwutviren, ihre herstellung und verwendung
US5427908A (en) 1990-05-01 1995-06-27 Affymax Technologies N.V. Recombinant library screening methods
GB9015198D0 (en) 1990-07-10 1990-08-29 Brien Caroline J O Binding substance
ES2139598T3 (es) 1990-07-10 2000-02-16 Medical Res Council Procedimientos para la produccion de miembros de parejas de union especifica.
EP0469897A3 (en) 1990-07-31 1992-11-19 The Wistar Institute Engineered antibodies
IE76732B1 (en) 1990-08-07 1997-11-05 Becton Dickinson Co One step test for absolute counts
US6255458B1 (en) 1990-08-29 2001-07-03 Genpharm International High affinity human antibodies and human antibodies against digoxin
US7041871B1 (en) 1995-10-10 2006-05-09 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
CA2089661C (en) 1990-08-29 2007-04-03 Nils Lonberg Transgenic non-human animals capable of producing heterologous antibodies
WO1993012227A1 (en) 1991-12-17 1993-06-24 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
US5770429A (en) 1990-08-29 1998-06-23 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
WO1992009690A2 (en) 1990-12-03 1992-06-11 Genentech, Inc. Enrichment method for variant proteins with altered binding properties
ES2330052T3 (es) 1991-03-01 2009-12-03 Dyax Corporation Proteina quimerica que comprende micro-proteinas que tienen dos o mas puentes disulfuro y relaizaciones de las mismas.
WO1992017566A2 (en) 1991-03-29 1992-10-15 Genentech, Inc. Methods for selection of recombinant host cells expressing high levels of a desired protein
ATE414768T1 (de) 1991-04-10 2008-12-15 Scripps Research Inst Bibliotheken heterodimerer rezeptoren mittels phagemiden
WO1994004679A1 (en) 1991-06-14 1994-03-03 Genentech, Inc. Method for making humanized antibodies
US5637481A (en) 1993-02-01 1997-06-10 Bristol-Myers Squibb Company Expression vectors encoding bispecific fusion proteins and methods of producing biologically active bispecific fusion proteins in a mammalian cell
DE4122599C2 (de) 1991-07-08 1993-11-11 Deutsches Krebsforsch Phagemid zum Screenen von Antikörpern
EP0552108B1 (en) 1992-01-17 1999-11-10 Lakowicz, Joseph R. Energy transfer phase-modulation fluoro-immunoassay
US5667988A (en) 1992-01-27 1997-09-16 The Scripps Research Institute Methods for producing antibody libraries using universal or randomized immunoglobulin light chains
US7067284B1 (en) 1992-01-27 2006-06-27 The Scripps Research Institute Methods for producing antibody libraries using universal or randomized immunoglobulin light chains
SE9201984D0 (sv) 1992-06-29 1992-06-29 Pharmacia Biosensor Ab Improvement in optical assays
DE69334095T2 (de) 1992-07-17 2007-04-12 Dana-Farber Cancer Institute, Boston Verfahren zur intrazellulären Bindung von zielgerichteten Molekülen
SG48760A1 (en) 1992-07-24 2003-03-18 Abgenix Inc Generation of xenogenetic antibodies
DK1167384T3 (da) 1992-10-28 2007-04-10 Genentech Inc Vaskular endotheliel cellevækstfaktor antagonister
EP0669986B1 (en) 1992-11-13 2003-04-09 Idec Pharmaceuticals Corporation Fully impaired consensus kozac sequences for mammalian expression
WO1994023046A1 (en) 1993-04-07 1994-10-13 The Government Of The United States Of America As Represented By The Secretary, Department Of Health And Human Services Dna sequence which acts as a chromatin insulator element to protect expressed genes from cis-acting regulatory sequences in mammalian cells
ATE204325T1 (de) 1993-04-29 2001-09-15 Unilever Nv Herstellung von antikörpern oder funktionstüchtig gemachten teilen davon, abgeleitet von schweren ketten von immunglobulinen von camelidae
US5693506A (en) 1993-11-16 1997-12-02 The Regents Of The University Of California Process for protein production in plants
US5827690A (en) 1993-12-20 1998-10-27 Genzyme Transgenics Corporatiion Transgenic production of antibodies in milk
DE69430014D1 (de) 1993-12-23 2002-04-04 Infigen Inc EMBRYONALE STAMMZELLEN von Huftieren ALS KERNDONATOREN UND KERNTRANSFERTECHNIKEN ZUR HERSTELLUNG CHIMÄRER UND TRANSGENER TIERE
PT1231268E (pt) 1994-01-31 2005-11-30 Univ Boston Bancos de anticorpos policlonais
FR2717187B1 (fr) 1994-03-10 1996-05-31 Transgene Sa Usage combiné de deux cassettes d'expression pour la production d'une protéine d'intérêt.
US6080560A (en) 1994-07-25 2000-06-27 Monsanto Company Method for producing antibodies in plant cells
JPH08116978A (ja) 1994-10-18 1996-05-14 Nisshinbo Ind Inc 抗体Ｆａｂライブラリーの作製法
EP0807173B1 (en) 1994-12-30 2007-08-08 Planet Biotechnology, Inc. Methods for producing immunoglobulins containing protection proteins in plants and their use
US5731168A (en) 1995-03-01 1998-03-24 Genentech, Inc. Method for making heteromultimeric polypeptides
US6265150B1 (en) 1995-06-07 2001-07-24 Becton Dickinson & Company Phage antibodies
US5783186A (en) 1995-12-05 1998-07-21 Amgen Inc. Antibody-induced apoptosis
US5885827A (en) 1996-01-23 1999-03-23 The Regents Of The Universtiy Of California Eukaryotic high rate mutagenesis system
US6300065B1 (en) 1996-05-31 2001-10-09 Board Of Trustees Of The University Of Illinois Yeast cell surface display of proteins and uses thereof
US6825396B2 (en) 1996-06-12 2004-11-30 Board Of Trustees Operating Michigan State University Methods for tissue specific synthesis of protein in eggs of transgenic hens
CA2267139A1 (en) 1996-10-08 1998-04-16 Ton Logtenberg Methods and means for selecting peptides and proteins having specific affinity for a target
GB9621113D0 (en) 1996-10-10 1996-11-27 Univ Southampton Transgenic fish
EP0963441A4 (en) 1996-12-02 2002-10-02 Univ Wake Forest INACTIVATION OF HIV CO-RECEPTORS FOR THE THERAPY OF HIV INFECTION
CA2278786C (en) 1997-01-21 2010-07-20 The General Hospital Corporation Selection of proteins using rna-protein fusions
BR9808192A (pt) 1997-03-06 2000-10-24 Infigen Inc Processo de clonagem de animais
CN1175111C (zh) 1997-03-14 2004-11-10 艾德药品公司 将基因整合至哺乳动物细胞特定位点的方法及所用载体
US6080912A (en) 1997-03-20 2000-06-27 Wisconsin Alumni Research Foundation Methods for creating transgenic animals
DK1325932T5 (da) 1997-04-07 2005-10-03 Genentech Inc Anti-VEGF antistoffer
US20030207346A1 (en) 1997-05-02 2003-11-06 William R. Arathoon Method for making multispecific antibodies having heteromultimeric and common components
US20020062010A1 (en) 1997-05-02 2002-05-23 Genentech, Inc. Method for making multispecific antibodies having heteromultimeric and common components
EP0983303B1 (en) 1997-05-21 2006-03-08 Biovation Limited Method for the production of non-immunogenic proteins
EP1017838A2 (en) 1997-09-23 2000-07-12 Oxford Biomedica (UK) Limited Expression of genes in hematopoietic stem cells in hischaemic conditions
GB9722131D0 (en) 1997-10-20 1997-12-17 Medical Res Council Method
EP1027439B1 (en) 1997-10-27 2010-03-17 Bac Ip B.V. Multivalent antigen-binding proteins
DE69823652D1 (de) 1997-11-26 2004-06-09 Mitsubishi Precision Co Ltd Informationsleitungssystem
IL138579A0 (en) 1998-03-30 2001-10-31 Res Dev Foundation Corticotropin releasing factor receptor 1-deficient mice
US6413776B1 (en) 1998-06-12 2002-07-02 Galapagos Geonomics N.V. High throughput screening of gene function using adenoviral libraries for functional genomics applications
GB9823930D0 (en) 1998-11-03 1998-12-30 Babraham Inst Murine expression of human ig\ locus
AU3475100A (en) 1999-01-29 2000-08-18 Imclone Systems Incorporated Antibodies specific to kdr and uses thereof
DK2067788T3 (da) 1999-05-18 2015-10-19 Dyax Corp Fab-fragmentbiblioteker og fremgangsmåder til anvendelse deraf
US6472147B1 (en) 1999-05-25 2002-10-29 The Scripps Research Institute Methods for display of heterodimeric proteins on filamentous phage using pVII and pIX, compositions, vectors and combinatorial libraries
US6833268B1 (en) 1999-06-10 2004-12-21 Abgenix, Inc. Transgenic animals for producing specific isotypes of human antibodies via non-cognate switch regions
MXPA01013458A (es) 1999-06-25 2002-07-30 Genentech Inc Anticuerpos humanizados anti-erbb2 y tratamiento con anticuerpos anti-erbb2.
WO2001019394A2 (en) 1999-09-15 2001-03-22 Therapeutic Human Polyclonals, Inc. Immunotherapy with substantially human polyclonal antibody preparations purified from genetically engineered birds
US6180357B1 (en) 1999-10-08 2001-01-30 Arius Research, Inc. Individualized patient-specific anti-cancer antibodies
AU7676300A (en) 1999-10-12 2001-04-23 Cambridge Antibody Technology Limited Human anti-adipocyte monoclonal antibodies and their use
JP2003513635A (ja) 1999-11-01 2003-04-15 カイロン コーポレイション 発現ベクター、トランスフェクション系、およびそれらの使用方法
GB9928787D0 (en) 1999-12-03 2000-02-02 Medical Res Council Direct screening method
ATE386747T1 (de) 1999-12-27 2008-03-15 Crucell Holland Bv Antikörper gegen ep-cam
CA2401965A1 (en) 2000-02-29 2001-09-07 Auburn University Production of antibodies in transgenic plastids
EP1284752A4 (en) 2000-04-26 2004-08-18 Elusys Therapeutics Inc BISPECIFIC MOLECULES AND USES THEREOF
AU6595301A (en) 2000-05-12 2001-11-20 Vlaams Interuniv Inst Biotech Use of inhibitors of placental growth factor for the treatment of pathological angiogenesis, pathological arteriogenesis, inflammation, tumour formation and/or vascular leakage
CA2419148C (en) 2000-05-16 2011-07-12 Thomas Jefferson University Rabies virus-specific neutralizing human monoclonal antibodies and nucleic acids and related methods
IL153567A0 (en) 2000-06-29 2003-07-06 Abbott Lab Dual specificity antibodies and methods of making and using
EP1184458A1 (en) 2000-08-28 2002-03-06 U-BISys B.V. Differentially expressed CD46 epitopes, proteinaceous molecules capable of binding thereto, and uses thereof
EP1188771A1 (en) 2000-09-15 2002-03-20 U-BISys B.V. Libraries of human heavy chain variable fragments in a functional format
US7737258B2 (en) 2000-10-18 2010-06-15 Sloan-Kettering Institute For Cancer Research Uses of monoclonal antibody 8H9
US7105348B2 (en) 2000-10-31 2006-09-12 Regeneron Pharmaceuticals, Inc. Methods of modifying eukaryotic cells
US6596541B2 (en) 2000-10-31 2003-07-22 Regeneron Pharmaceuticals, Inc. Methods of modifying eukaryotic cells
ES2405944T3 (es) 2000-11-30 2013-06-04 Medarex, Inc. Ácidos nucleicos que codifican las secuencias de inmunoglobulina humana reorganizadas a partir de ratones transcromoscómicos transgénicos zadas
FR2817875B1 (fr) 2000-12-07 2005-03-18 Technopharm Procede de preparation d'un anticorps monoclonal humain, de fragments de celui-ci ou d'anticorps comprenant de tels fragments, les anticorps ainsi obtenus et leur utilisation
US7838287B2 (en) 2001-01-25 2010-11-23 Evolva Sa Library of a collection of cells
CA2435972C (en) 2001-01-26 2011-09-13 University Of Lausanne Matrix attachment regions and methods for use thereof
AU2002327164A1 (en) 2001-01-29 2002-12-09 Idec Pharmaceuticals Corporation Engineered tetravalent antibodies and methods of use
EP1379125A4 (en) 2001-03-22 2004-12-08 Abbott Gmbh & Co Kg TRANSGENIC ANIMALS THAT EXPRESS SPECIFIC ANTIBODIES FOR INTERESTING GENES AND THEIR USE
GB0110029D0 (en) 2001-04-24 2001-06-13 Grosveld Frank Transgenic animal
US7595378B2 (en) 2001-06-13 2009-09-29 Genmab A/S Human monoclonal antibodies to epidermal growth factor receptor (EGFR)
AU2002345673B2 (en) 2001-06-13 2007-04-26 Genmab A/S Human monoclonal antibodies to epidermal growth factor receptor (EGFR)
DK1402025T3 (da) 2001-06-15 2006-06-06 Crucell Holland Bv Kimære fager
WO2004003211A1 (en) 2002-06-26 2004-01-08 Imclone Systems Incorporated Bispecific antibodies that bind to vegf receptors
DE60237282D1 (de) 2001-06-28 2010-09-23 Domantis Ltd Doppelspezifischer ligand und dessen verwendung
WO2003016501A2 (en) 2001-08-21 2003-02-27 Thomas Jefferson University Recombinant antibodies, and compositions and methods for making and using the same
KR100968664B1 (ko) 2001-08-27 2010-07-06 제넨테크, 인크. 항체 발현 및 조립을 위한 시스템
EP2022799A2 (en) 2001-11-16 2009-02-11 Biogen Idec Inc. Polycistronic expression of antibodies
AU2002363861A1 (en) 2001-11-30 2003-06-10 Crucell Holland B.V. Antigen presenting cell targeting conjugate, an intigen presenting cell contacted with such conjugate, their use for vaccination or as medicament, and methods for their production or generation
ATE394673T1 (de) 2001-11-30 2008-05-15 Ca Nat Research Council Selbstanordnende moleküle
GB0130267D0 (en) 2001-12-19 2002-02-06 Neutec Pharma Plc Focussed antibody technology
US7244592B2 (en) 2002-03-07 2007-07-17 Dyax Corp. Ligand screening and discovery
GB2387030A (en) 2002-03-26 2003-10-01 Thales Plc Compensation of mutual coupling in array antenna systems
EP1510943A4 (en) 2002-05-31 2007-05-09 Celestar Lexico Sciences Inc INTERACTION PREDICTION DEVICE
CA2488441C (en) 2002-06-03 2015-01-27 Genentech, Inc. Synthetic antibody phage libraries
WO2003106674A2 (en) 2002-06-14 2003-12-24 Chromagenics B.V. Means and methods for regulating gene expression
AU2003238719B2 (en) 2002-06-14 2008-02-21 Chromagenics B.V. A method for simultaneous production of multiple proteins, vectors and cells for use therein
US20080241166A1 (en) 2002-06-28 2008-10-02 Domantis Limited Ligands that bind a receptor
CA2492067A1 (fr) 2002-07-10 2004-01-22 Agostino Di Trapani Element de construction
EP1439234A1 (en) 2003-01-08 2004-07-21 ARTEMIS Pharmaceuticals GmbH Targeted transgenesis using the rosa26 locus
EP2395017A3 (en) 2003-05-30 2012-12-19 Merus B.V. Design and use of paired variable regions of specific binding molecules
US20100069614A1 (en) 2008-06-27 2010-03-18 Merus B.V. Antibody producing non-human mammals
NZ543635A (en) 2003-06-25 2008-05-30 Crucell Holland Bv Human C-type lectin: a suitable target molecule for binding molecules, particularly immunoconjugates, in the diagnosis, prevention and/or treatment of myeloid neoplastic diseases such as AML and CML
KR101206206B1 (ko) 2003-07-22 2012-11-29 크루셀 홀란드 비.브이. 사스-코로나바이러스에 대한 결합분자 및 그것의 용도
ES2378767T3 (es) 2003-12-23 2012-04-17 Crucell Holland B.V. Molécula de unión humana contra CD1a
DK1737971T3 (da) 2004-01-20 2017-11-13 Merus Nv Blandinger af bindingsproteiner
CN102139106B (zh) 2004-05-27 2014-10-08 克鲁塞尔荷兰公司 能中和狂犬病病毒的结合分子及其应用
EP1753783B1 (en) 2004-06-03 2014-08-06 Novimmune SA Anti-cd3 antibodies and methods of use thereof
JP2008511337A (ja) 2004-09-02 2008-04-17 ジェネンテック・インコーポレーテッド ヘテロ多量体分子
JP4487068B2 (ja) 2004-10-12 2010-06-23 国立大学法人 岡山大学 細胞の遺伝子変異機能の制御による変異タンパク質の作製方法
JP4852046B2 (ja) 2004-10-12 2012-01-11 クルセル ホランド ベー ヴェー 癌の治療及び検出に用いる結合分子
ATE550037T1 (de) 2004-11-11 2012-04-15 Crucell Holland Bv Zusammensetzungen gegen das sars-coronavirus und ihre verwendungen
EP1874817A2 (en) 2005-04-29 2008-01-09 Innate Pharma Transgenic animals and methods of making recombinant antibodies
WO2006120230A2 (en) 2005-05-12 2006-11-16 Crucell Holland B.V. Host cell specific binding molecules capable of neutralizing viruses and uses thereof
EP1893645A1 (en) 2005-06-23 2008-03-05 Crucell Holland B.V. Optimization of west nile virus antibodies
WO2007031550A2 (en) 2005-09-15 2007-03-22 Crucell Holland B.V. Method for preparing immunoglobulin libraries
CN101370526B (zh) * 2005-11-28 2013-10-09 根马布股份公司 重组单价抗体及其制备方法
KR101866623B1 (ko) 2005-11-28 2018-07-04 젠맵 에이/에스 재조합 １가 항체 및 그의 제조 방법
EP2505058A1 (en) 2006-03-31 2012-10-03 Medarex, Inc. Transgenic animals expressing chimeric antibodies for use in preparing human antibodies
NZ573557A (en) 2006-06-02 2010-08-27 Regeneron Pharma High affinity antibodies to human il-6 receptor
JP5586952B2 (ja) 2006-06-06 2014-09-10 クルセル ホランド ベー ヴェー 腸球菌に対する殺活性を有するヒトの結合分子及びその使用方法
EP3018142A1 (en) 2006-06-06 2016-05-11 Crucell Holland B.V. Human binding molecules having killing activity against staphylococci and uses thereof
ES2399075T3 (es) 2006-08-30 2013-03-25 Genentech, Inc. Anticuerpos multiespecíficos
MY170607A (en) 2006-09-07 2019-08-20 Crucell Holland Bv Human binding molecules capable of neutralizing influenza virus h5n1 and uses thereof
DK2769992T3 (da) 2006-10-02 2021-03-22 Regeneron Pharma Humane antistoffer med høj affinitet for human IL-4-receptor
US8290739B2 (en) 2006-10-20 2012-10-16 Amfit, Inc. Method for determining relative mobility of regions of an object
NO347649B1 (no) 2006-12-14 2024-02-12 Regeneron Pharma Humant antistoff eller antistoff fragment som spesifikt binder human deltaliknende ligand 4 (hDII4), nukleinsyremolekyl som koder for slike og vektor og vert-vektorsystemer, samt fremgangsmåte for fremstilling, sammensetning og anvendelse.
CA2681974C (en) 2007-03-29 2019-12-31 Genmab A/S Bispecific antibodies and methods for production thereof
ITMI20071522A1 (it) 2007-07-27 2009-01-28 Areta Internat S R L Vaccino idiotipico
WO2009023270A2 (en) 2007-08-15 2009-02-19 Amunix, Inc. Compositions and methods for modifying properties of biologically active polypeptides
EP3753947A1 (en) 2007-09-14 2020-12-23 Adimab, LLC Rationally designed, synthetic antibody libraries and uses therefor
US8536310B2 (en) 2007-10-17 2013-09-17 Arca Biopharma, Inc. Antibodies to CLL-1
CA2715043C (en) 2008-02-05 2021-02-16 Zymeworks Inc. Methods for determining correlated residues in a protein or other biopolymer using molecular dynamics
EP2556747B1 (en) 2008-06-27 2020-12-02 Merus N.V. Antibody producing transgenic mice
NZ594665A (en) 2009-03-20 2013-08-30 Genentech Inc Bispecific anti-her antibodies
US8470327B2 (en) 2009-05-11 2013-06-25 Crucell Holland B.V. Human binding molecules capable of neutralizing influenza virus H3N2 and uses thereof
SI2975051T1 (sl) 2009-06-26 2021-08-31 Regeneron Pharmaceuticals, Inc. Zlahka izolirana bispecifična protitelesa z nativnim imunoglobulinskim formatom
US8765900B2 (en) 2009-08-27 2014-07-01 Bayer Materialscience Llc Aliphatic moisture-curable resins, coating compositions, and related processes
US9493578B2 (en) * 2009-09-02 2016-11-15 Xencor, Inc. Compositions and methods for simultaneous bivalent and monovalent co-engagement of antigens
CA2781539C (en) 2009-11-23 2021-07-20 Amgen Inc. Monomeric antibody fc
US20120021409A1 (en) 2010-02-08 2012-01-26 Regeneron Pharmaceuticals, Inc. Common Light Chain Mouse
US9527926B2 (en) * 2010-05-14 2016-12-27 Rinat Neuroscience Corp. Heterodimeric proteins and methods for producing and purifying them
EP2603526A1 (en) * 2010-08-13 2013-06-19 Medimmune Limited Monomeric polypeptides comprising variant fc regions and methods of use
PL2606064T3 (pl) 2010-08-16 2015-07-31 Novimmune Sa Sposoby wytwarzania wieloswoistych i wielowartościowych przeciwciał
RS59589B1 (sr) * 2010-11-05 2019-12-31 Zymeworks Inc Dizajniranje stabilnog heterodimernog antitela sa mutacijama u fc domenu
SI2738258T2 (sl) 2011-02-25 2023-05-31 Regeneron Pharmaceuticals, Inc. Miši ADAM6
SG10201602371VA (en) 2011-03-25 2016-04-28 Glenmark Pharmaceuticals Sa Hetero-dimeric immunoglobulins
JP2013004215A (ja) 2011-06-14 2013-01-07 Hitachi Ltd リチウムイオン二次電池
CA2791109C (en) 2011-09-26 2021-02-16 Merus B.V. Generation of binding molecules
JP2015509380A (ja) 2012-03-06 2015-03-30 リジェネロン・ファーマシューティカルズ・インコーポレイテッドＲｅｇｅｎｅｒｏｎ Ｐｈａｒｍａｃｅｕｔｉｃａｌｓ， Ｉｎｃ． 共通の軽鎖のマウス
CA2870821C (en) 2012-04-20 2022-06-14 Merus B.V. Methods and means for the production of ig-like molecules
SG11201407644UA (en) 2012-06-05 2014-12-30 Regeneron Pharma Methods for making fully human bispecific antibodies using a common light chain
LT2900694T (lt) 2012-09-27 2018-11-12 Merus N.V. Bispecifiniai igg antikūnai kaip t ląstelių aktyvatoriai
UY35148A (es) * 2012-11-21 2014-05-30 Amgen Inc Immunoglobulinas heterodiméricas
US10798961B2 (en) 2013-03-15 2020-10-13 Altria Client Services Llc Functional food and beverage compositions with improved taste through the use of sensates
PL3115376T3 (pl) 2015-07-10 2019-01-31 Merus N.V. Ludzkie przeciwciało wiążące cd3

Patent Citations (23)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0120694A2 (en) 1983-03-25 1984-10-03 Celltech Limited Processes for the production of multichain polypeptides or proteins
US4816567A (en) 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
EP0187045A2 (en) 1984-12-28 1986-07-09 Elf Atochem S.A. Radiation curable macromonomer printing inks
US4801687A (en) 1986-10-27 1989-01-31 Bioprobe International, Inc. Monoclonal antibody purification process using protein A
EP0314161A1 (en) 1987-10-28 1989-05-03 Bristol-Myers Squibb Company Human immunoglobulines produced by recombinant DNA techniques
US5151504A (en) 1989-11-17 1992-09-29 E. R. Squibb & Sons, Inc. Method for purification of monoclonal antibodies
EP0481790A2 (en) 1990-10-17 1992-04-22 The Wellcome Foundation Limited Antibody production
EP0523949A1 (en) 1991-07-15 1993-01-20 The Wellcome Foundation Limited Production of antibodies
WO1998050431A2 (en) 1997-05-02 1998-11-12 Genentech, Inc. A method for making multispecific antibodies having heteromultimeric and common components
WO2000063403A2 (en) 1999-04-15 2000-10-26 Crucell Holland B.V. Recombinant protein production in a human cell using sequences encoding adenovirus e1 protein
WO2003004704A2 (en) 2001-07-04 2003-01-16 Chromagenics B.V. Method of selecting dna sequence with transcription modulating activity using a vector comprising an element with a gene transcription repressing activity
WO2004009618A2 (en) 2002-07-18 2004-01-29 Crucell Holland B.V. Recombinant production of mixtures of antibodies
WO2004061104A2 (en) 2003-01-07 2004-07-22 Symphogen A/S Method for manufacturing recombinant polyclonal proteins
US20100015133A1 (en) 2005-03-31 2010-01-21 Chugai Seiyaku Kabushiki Kaisha Methods for Producing Polypeptides by Regulating Polypeptide Association
WO2007110205A2 (en) 2006-03-24 2007-10-04 Merck Patent Gmbh Engineered heterodimeric protein domains
WO2007147901A1 (en) 2006-06-22 2007-12-27 Novo Nordisk A/S Production of bispecific antibodies
WO2009080252A1 (en) 2007-12-21 2009-07-02 F. Hoffmann-La Roche Ag Bivalent, bispecific antibodies
WO2009080253A1 (en) 2007-12-21 2009-07-02 F. Hoffmann-La Roche Ag Bivalent, bispecific antibodies
WO2009080251A1 (en) 2007-12-21 2009-07-02 F. Hoffmann-La Roche Ag Bivalent, bispecific antibodies
WO2009089004A1 (en) 2008-01-07 2009-07-16 Amgen Inc. Method for making antibody fc-heterodimeric molecules using electrostatic steering effects
WO2010084197A1 (en) * 2009-01-26 2010-07-29 Genmab A/S Methods for producing mixtures of antibodies
WO2010129304A2 (en) 2009-04-27 2010-11-11 Oncomed Pharmaceuticals, Inc. Method for making heteromultimeric molecules
US20110195454A1 (en) * 2010-02-08 2011-08-11 Regeneron Pharmaceuticals, Inc. Common Light Chain Mouse

Non-Patent Citations (27)

* Cited by examiner, † Cited by third party
Title
"Mammalian Cell Biotechnology : a Practical Approach", 1991, IRL PRESS
"Mammalian Cell Biotechnology: a Practical Approach", 1991, IRL PRESS
"Tissue Culture", 1973, ACADEMIC PRESS
CAPELLE, M.A.H.; BRUGGER, P.; ARVINTE, T, VACCINE, vol. 23, 2005, pages 1686 - 1694
COLIGAN JE ET AL., CURRENT PROTOCOLS IN PROTEIN SCIENCE, 1995
COLIGAN JE; DUNN BM; PLOEGH HL; SPEICHER DW; WINGFIELD PT, CURRENT PROTOCOLS IN PROTEIN SCIENCE, 1995
DAVIS JH ET AL., PROTEIN ENGINEERING, DESIGN & SELECTION, 2010, pages 195 - 202
DAVIS JH ET AL.: "Protein Engineering", DESIGN & SELECTION, 2010, pages 195 - 202
DE KRUIF ET AL., BIOTECH BIOENG, 2010
DE VRIES, NATURE PROTOCOLS, 2010, pages 883
DEISENHOFER J, BIOCHEMISTRY, 1981, pages 2361 - 2370
DEISENHOFER J., BIOCHEMISTRY, 1981, pages 2361 - 2370
DEMEULE, B.; GURNY, R.; ARVINTE, T., INT. J. PHARM, vol. 329, 2007, pages 37 - 45
DEMEULE, B.; LAWRENCE, M.J.; DRAKE, A.F.; GURNY, R.; ARVINTE, T, BIOCHIM. BIOPHYS. ACTA, vol. 1774, 2007, pages 146 - 153
ELLERSON JR. ET AL., J. IMMUNOL, 1976, pages 510 - 517
GUNASEKARAN ET AL., J.BIOL.CHEM., 2010, pages 19637 - 19646
IDUSOGIE, E.E. ET AL., J.I., 2000, pages 4178 - 4184, Retrieved from the Internet <URL:httya://www.rcsb.org/pdb/explore/explore.do?structureId=1l6x>
IONESCU ET AL., J. PHARM. SCI., 2008, pages 1414
LAKOWICZ, J.R.: "Principles of fluorescence spectroscopy", 2006, KLUWER ACADEMIC/PLENUM PUBLISHERS
LEE; RICHARDS, J. MOL. BIOL., 1971, pages 379
MERCHANT, NATURE BIOTECHNOLOGY, 1998, pages 677
MILLER S., J. MOL. BIOL., 1990, pages 965 - 973
PADLAN, ADVANCES IN PROTEIN CHEMISTRY, 1996, pages 57 - 133
PAPADEA; CHECK, CRIT REV CLIN LAB SCI, vol. 27, no. 1, 1989, pages 27 - 58
RIDGEWAY, PROTEIN ENGINEERING, 1996, pages 617 - 621
STRELKAUSKAS A J ET AL: "HUMAN MONOCLONAL ANTIBODY: 2. SIMULTANEOUS EXPRESSION OF IGG AND IGM WITH SIMILAR BINDING SPECIFICITIES BY A HUMAN HYBRID CLONE", HYBRIDOMA, LIEBERT, NEW YORK, NY, US, vol. 6, no. 5, 1 January 1987 (1987-01-01), pages 479 - 488, XP009033431, ISSN: 0272-457X *
TAHALLAH ET AL., RCM, 2001

Cited By (334)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11230697B2 (en) 2006-09-01 2022-01-25 Therapeutic Human Polyclonals Inc. Enhanced expression of human or humanized immunoglobulin in non-human transgenic animals
US10927163B2 (en) 2007-12-21 2021-02-23 Hoffmann-La Roche, Inc. Bivalent, bispecific antibodies
US10138293B2 (en) 2007-12-21 2018-11-27 Hoffmann-La Roche, Inc. Bivalent, bispecific antibodies
US11993642B2 (en) 2009-04-07 2024-05-28 Hoffmann-La Roche Inc. Trivalent, bispecific antibodies
US9890204B2 (en) 2009-04-07 2018-02-13 Hoffmann-La Roche Inc. Trivalent, bispecific antibodies
US11673945B2 (en) 2009-06-16 2023-06-13 Hoffmann-La Roche Inc. Bispecific antigen binding proteins
US9676845B2 (en) 2009-06-16 2017-06-13 Hoffmann-La Roche, Inc. Bispecific antigen binding proteins
US10640555B2 (en) 2009-06-16 2020-05-05 Hoffmann-La Roche Inc. Bispecific antigen binding proteins
US9994646B2 (en) 2009-09-16 2018-06-12 Genentech, Inc. Coiled coil and/or tether containing protein complexes and uses thereof
US10106600B2 (en) 2010-03-26 2018-10-23 Roche Glycart Ag Bispecific antibodies
US10793621B2 (en) 2011-02-28 2020-10-06 Hoffmann-La Roche Inc. Nucleic acid encoding dual Fc antigen binding proteins
US10611825B2 (en) 2011-02-28 2020-04-07 Hoffmann La-Roche Inc. Monovalent antigen binding proteins
US9982036B2 (en) 2011-02-28 2018-05-29 Hoffmann-La Roche Inc. Dual FC antigen binding proteins
US10752664B2 (en) 2011-04-08 2020-08-25 Amgen Inc. Method of treating or ameliorating metabolic disorders using growth differentiation factor 15 (GDF-15)
US11639397B2 (en) 2011-08-23 2023-05-02 Roche Glycart Ag Bispecific antibodies specific for T-cell activating antigens and a tumor antigen and methods of use
US20140378665A1 (en) * 2012-01-26 2014-12-25 Amgen Inc. Growth differentiation factor 15 (gdf-15) polypeptides
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US9714276B2 (en) * 2012-01-26 2017-07-25 Amgen Inc. Growth differentiation factor 15 (GDF-15) polypeptides
US11739131B2 (en) 2012-01-26 2023-08-29 Amgen Inc. Growth differentiation factor 15 (GDF-15) polypeptides
US10336798B2 (en) 2012-01-26 2019-07-02 Amgen Inc. Growth differentiation factor 15 (GDF-15) polypeptides
US11926859B2 (en) 2012-04-20 2024-03-12 Merus N.V. Methods and means for the production of Ig-like molecules
US12123043B2 (en) 2012-04-20 2024-10-22 Merus N.V. Methods and means for the production of Ig-like molecules
US10087250B2 (en) 2012-10-08 2018-10-02 Roche Glycart Ag Fc-free antibodies comprising two fab-fragments and methods of use
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US10155815B2 (en) 2013-02-26 2018-12-18 Roche Glycart Ag Bispecific T cell activating antigen binding molecules
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EP4026848A1 (en) 2015-12-09 2022-07-13 F. Hoffmann-La Roche AG Type ii anti-cd20 antibody for reducing the cytokine release syndrome
EP3178848A1 (en) 2015-12-09 2017-06-14 F. Hoffmann-La Roche AG Type ii anti-cd20 antibody for reducing formation of anti-drug antibodies
US10596257B2 (en) 2016-01-08 2020-03-24 Hoffmann-La Roche Inc. Methods of treating CEA-positive cancers using PD-1 axis binding antagonists and anti-CEA/anti-CD3 bispecific antibodies
EP3862365A1 (en) 2016-01-08 2021-08-11 F. Hoffmann-La Roche AG Methods of treating cea-positive cancers using pd-1 axis binding antagonists and anti-cea/anti-cd3 bispecific antibodies
WO2017121877A1 (en) 2016-01-13 2017-07-20 Genmab A/S Axl-specific antibody-drug conjugates for cancer treatment
US11242390B2 (en) 2016-03-22 2022-02-08 Hoffmann-La Roche Inc. Protease-activated T cell bispecific molecules
WO2017186853A1 (en) * 2016-04-27 2017-11-02 Bcrt Holding Bv T cell-based bi-specific immunotherapeutics
WO2017191101A1 (en) 2016-05-02 2017-11-09 F. Hoffmann-La Roche Ag The contorsbody - a single chain target binder
EP3889175A1 (en) 2016-05-02 2021-10-06 F. Hoffmann-La Roche AG The contorsbody - a single chain target binder
US11130808B2 (en) 2016-05-26 2021-09-28 Qilu Puget Sound Biotherapeutics Corporation Mixtures of antibodies
EP3463451A1 (en) * 2016-05-26 2019-04-10 Qilu Puget Sound Biotherapeutics Corporation Mixtures of antibodies
EP3252078A1 (en) 2016-06-02 2017-12-06 F. Hoffmann-La Roche AG Type ii anti-cd20 antibody and anti-cd20/cd3 bispecific antibody for treatment of cancer
WO2018007314A1 (en) 2016-07-04 2018-01-11 F. Hoffmann-La Roche Ag Novel antibody format
WO2018011421A1 (en) 2016-07-14 2018-01-18 Genmab A/S Multispecific antibodies against cd40 and cd137
US11685786B2 (en) 2016-09-23 2023-06-27 Merus N.V. Binding molecules that bind CD137 and PD-L1
EP4342912A2 (en) 2016-09-23 2024-03-27 Merus N.V. Binding molecules that modulate a biological activity expressed by a cell
US11498977B2 (en) 2016-09-29 2022-11-15 Beijing Hanmi Pharmaceutical Co., Ltd. Heterodimeric immunoglobulin constructs and preparation methods thereof
WO2018059502A1 (en) * 2016-09-29 2018-04-05 Beijing Hanmi Pharmaceutical Co., Ltd. Heterodimeric immunoglobulin constructs and preparation methods thereof
IL265605B2 (en) * 2016-09-29 2024-06-01 Beijing Hanmi Pharmaceutical Co Ltd Heterodimeric immunoglobulin constructs, pharmaceutical composition comprising the same, use and preparation methods thereof
IL265605B1 (en) * 2016-09-29 2024-02-01 Beijing Hanmi Pharmaceutical Co Ltd Heterodimeric immunoglobulin structures, a pharmaceutical preparation containing them, their use and methods for their preparation
WO2018060301A1 (en) 2016-09-30 2018-04-05 F. Hoffmann-La Roche Ag Bispecific antibodies against cd3
US10882918B2 (en) 2016-09-30 2021-01-05 Hoffmann-La Roche Inc. Bispecific T cell activating antigen binding molecules
AU2017345454B2 (en) * 2016-10-19 2024-10-31 Invenra Inc. Antibody constructs
WO2018075692A3 (en) * 2016-10-19 2019-04-04 Invenra Inc. Antibody constructs
WO2018083126A1 (en) 2016-11-01 2018-05-11 Genmab B.V. Polypeptide variants and uses thereof
US11319378B2 (en) 2016-11-18 2022-05-03 Beijing Hanmi Pharmaceutical Co., Ltd. Anti-PD-1/anti-HER2 natural antibody structural heterodimeric bispecific antibody and method of preparing the same
US11981737B2 (en) 2016-11-18 2024-05-14 Hoffmann-La Roche Inc. Anti-HLA-G antibodies and use thereof
WO2018146317A1 (en) 2017-02-10 2018-08-16 Genmab B.V. Polypeptide variants and uses thereof
WO2018162749A1 (en) 2017-03-09 2018-09-13 Genmab A/S Antibodies against pd-l1
EP4527854A2 (en) 2017-03-09 2025-03-26 Genmab A/S Antibodies against pd-l1
US11780925B2 (en) 2017-03-31 2023-10-10 Merus N.V. ErbB-2 and ErbB3 binding bispecific antibodies for use in the treatment of cells that have an NRG1 fusion gene
WO2018178396A1 (en) 2017-03-31 2018-10-04 Genmab Holding B.V. Bispecific anti-cd37 antibodies, monoclonal anti-cd37 antibodies and methods of use thereof
US12247078B2 (en) 2017-03-31 2025-03-11 Merus N.V. ERBB-2 and ERBB-3 binding bispecific antibodies for use in the treatment of cells that have an NRG1 fusion gene
US11685790B2 (en) 2017-04-03 2023-06-27 Hoffmann-La Roche Inc. Antibodies binding to STEAP-1
US12129306B2 (en) 2017-04-03 2024-10-29 Hoffman-La Roche Inc. Antibodies binding to STEAP-1
WO2018184966A1 (en) 2017-04-03 2018-10-11 F. Hoffmann-La Roche Ag Antibodies binding to steap-1
WO2018184965A1 (en) 2017-04-03 2018-10-11 F. Hoffmann-La Roche Ag Immunoconjugates of il-2 with an anti-pd-1 and tim-3 bispecific antibody
WO2018184964A1 (en) 2017-04-03 2018-10-11 F. Hoffmann-La Roche Ag Immunoconjugates of an anti-pd-1 antibody with a mutant il-2 or with il-15
US11180571B2 (en) 2017-04-03 2021-11-23 Hoffmann-La Roche Inc. Antibodies binding to STEAP-1
EP4516809A2 (en) 2017-04-05 2025-03-05 F. Hoffmann-La Roche AG Bispecific antibodies specifically binding to pd1 and lag3
WO2018185043A1 (en) 2017-04-05 2018-10-11 F. Hoffmann-La Roche Ag Bispecific antibodies specifically binding to pd1 and lag3
WO2018189220A1 (en) 2017-04-13 2018-10-18 F. Hoffmann-La Roche Ag An interleukin-2 immunoconjugate, a cd40 agonist, and optionally a pd-1 axis binding antagonist for use in methods of treating cancer
US12195551B2 (en) 2017-05-17 2025-01-14 Merus N.V. Combination of an ErbB-2/ErbB-3 bispecific antibody with endocrine therapy for breast cancer
WO2018220099A1 (en) 2017-06-02 2018-12-06 F. Hoffmann-La Roche Ag Type ii anti-cd20 antibody and anti-cd20/cd3 bispecific antibody for treatment of cancer
WO2018224609A1 (en) 2017-06-07 2018-12-13 Genmab B.V. Therapeutic antibodies based on mutated igg hexamers
EP3638700A4 (en) * 2017-06-14 2021-04-21 Dingfu Biotarget Co., Ltd HETERODIMER PROTEIN AND USE OF IT
US11845806B2 (en) 2017-06-14 2023-12-19 Dingfu Biotarget Co., Ltd. Proteinaceous heterodimer and use thereof
US11753470B2 (en) 2017-07-06 2023-09-12 Merus N.V. Bispecific anti PD1-anti TIM3 antibodies
US11732043B2 (en) 2017-07-06 2023-08-22 Merus N.V. Antibodies that modulate a biological activity expressed by a cell
AU2018296067B2 (en) * 2017-07-06 2021-06-10 Merus N.V. Binding molecules that modulate a biological activity expressed by a cell
WO2019009727A1 (en) 2017-07-06 2019-01-10 Merus N.V. ANTI-PD1 ANTI-TIM3 BISPECIFIC ANTIBODIES
US11667714B2 (en) 2017-07-06 2023-06-06 Merus N.V. Binding molecules that modulate a biological activity expressed by a cell
WO2019009728A1 (en) 2017-07-06 2019-01-10 Merus N.V. ANTIBODIES THAT MODULATE A BIOLOGICAL ACTIVITY EXPRESSED BY A CELL
WO2019009726A1 (en) 2017-07-06 2019-01-10 Merus N.V. BINDING MOLECULES FOR MODULATING THE BIOLOGICAL ACTIVITY EXPRESSED BY A CELL
WO2019025545A1 (en) 2017-08-04 2019-02-07 Genmab A/S BINDING AGENTS BINDING TO PD-L1 AND CD137 AND THEIR USE
US11773170B2 (en) 2017-08-09 2023-10-03 Merus N.V. Antibodies that bind EGFR and cMET
WO2019031965A1 (en) 2017-08-09 2019-02-14 Merus N.V. ANTIBODIES THAT BIND EGFR AND CMET
EP4553088A2 (en) 2017-08-09 2025-05-14 Merus N.V. Antibodies that bind egfr and cmet
WO2019070047A1 (ja) 2017-10-06 2019-04-11 小野薬品工業株式会社 二重特異性抗体
WO2019077092A1 (en) 2017-10-20 2019-04-25 F. Hoffmann-La Roche Ag METHOD FOR GENERATING MULTISPECIFIC ANTIBODIES FROM MONOSPECIFIC ANTIBODIES
WO2019086362A1 (en) 2017-10-30 2019-05-09 F. Hoffmann-La Roche Ag Method for in vivo generation of multispecific antibodies from monospecific antibodies
US12043651B2 (en) 2017-11-01 2024-07-23 Hoffmann-La Roche Inc. Compbody—a multivalent target binder
WO2019086395A1 (en) 2017-11-01 2019-05-09 F. Hoffmann-La Roche Ag Trifab-contorsbody
WO2019086394A1 (en) 2017-11-01 2019-05-09 F. Hoffmann-La Roche Ag The compbody - a multivalent target binder
US12098213B2 (en) 2017-12-21 2024-09-24 Hoffmann-La Roche Inc. Antibodies binding to HLA-A2/WT1
WO2019122052A2 (en) 2017-12-21 2019-06-27 F. Hoffmann-La Roche Ag Antibodies binding to hla-a2/wt1
US11192957B2 (en) 2017-12-21 2021-12-07 Hoffmann-La Roche Inc. Antibodies binding to HLA-A2/WT1
WO2019145455A1 (en) 2018-01-24 2019-08-01 Genmab B.V. Polypeptide variants and uses thereof
WO2019154776A1 (en) 2018-02-06 2019-08-15 F. Hoffmann-La Roche Ag Treatment of ophthalmologic diseases
US11753471B2 (en) 2018-02-08 2023-09-12 Beijing Hanmi Pharmaceutical Co., Ltd. Anti-PD-1/anti-HER2 natural antibody structural heterodimeric bispecific antibody and method of preparing same
US11866498B2 (en) 2018-02-08 2024-01-09 Genentech, Inc. Bispecific antigen-binding molecules and methods of use
US12297270B2 (en) 2018-02-08 2025-05-13 Genentech, Inc. Bispecific antigen-binding molecules and methods of use
WO2019156199A1 (ja) 2018-02-09 2019-08-15 小野薬品工業株式会社 二重特異性抗体
WO2019154890A1 (en) 2018-02-09 2019-08-15 F. Hoffmann-La Roche Ag Antibodies binding to gprc5d
WO2019190327A2 (en) 2018-03-30 2019-10-03 Merus N.V. Multivalent antibody
CN111936514A (zh) * 2018-03-30 2020-11-13 美勒斯公司 多价抗体
WO2019190327A3 (en) * 2018-03-30 2019-11-14 Merus N.V. Multivalent antibody
US11952424B2 (en) 2018-03-30 2024-04-09 Merus N.V. Multivalent antibody
US11161889B2 (en) 2018-04-09 2021-11-02 Amgen Inc. Growth differentiation factor 15 fusion proteins
US12054527B2 (en) 2018-04-09 2024-08-06 Amgen Inc. Growth differentiation factor 15 fusion proteins
WO2019202041A1 (en) 2018-04-18 2019-10-24 F. Hoffmann-La Roche Ag Multispecific antibodies and use thereof
US12157770B2 (en) 2018-04-18 2024-12-03 Hoffmann-La Roche Inc. Anti-HLA-G antibodies and methods of use thereof
WO2019211472A1 (en) 2018-05-03 2019-11-07 Genmab B.V. Antibody variant combinations and uses thereof
WO2019232484A1 (en) 2018-06-01 2019-12-05 Compugen Ltd Anti-pvrig/anti-tigit bispecific antibodies and methods of use
US12173083B2 (en) 2018-06-09 2024-12-24 Boehringer Ingelheim International Gmbh Multi-specific binding proteins for cancer treatment
US11332541B2 (en) 2018-06-09 2022-05-17 Boehringer Ingelheim International Gmbh Multi-specific binding proteins for cancer treatment
WO2019243636A1 (en) 2018-06-22 2019-12-26 Genmab Holding B.V. Anti-cd37 antibodies and anti-cd20 antibodies, compositions and methods of use thereof
WO2020012038A1 (en) 2018-07-13 2020-01-16 Genmab A/S Trogocytosis-mediated therapy using cd38 antibodies
WO2020012036A1 (en) 2018-07-13 2020-01-16 Genmab A/S Variants of cd38 antibody and uses thereof
WO2020060405A1 (en) 2018-09-19 2020-03-26 Lava Therapeutics B.V. Dual acting cd1d immunoglobulin
WO2020060406A1 (en) 2018-09-19 2020-03-26 Lava Therapeutics B.V. Novel bispecific antibodies for use in the treatment of hematological malignancies
WO2020070313A1 (en) 2018-10-04 2020-04-09 Genmab Holding B.V. Pharmaceutical compositions comprising bispecific anti-cd37 antibodies
WO2020089051A1 (en) 2018-10-29 2020-05-07 F. Hoffmann-La Roche Ag Antibody formulation
WO2020094744A1 (en) 2018-11-06 2020-05-14 Genmab A/S Antibody formulation
WO2020127619A1 (en) 2018-12-21 2020-06-25 F. Hoffmann-La Roche Ag Antibodies binding to cd3
WO2020136060A1 (en) 2018-12-28 2020-07-02 F. Hoffmann-La Roche Ag A peptide-mhc-i-antibody fusion protein for therapeutic use in a patient with amplified immune response
WO2020141974A1 (en) 2018-12-31 2020-07-09 Merus N.V. Truncated multivalent multimers
WO2020141973A1 (en) 2018-12-31 2020-07-09 Merus N.V. Mixed binding domains
NL2022494B1 (en) 2019-02-01 2020-08-19 Lava Therapeutics B V Novel CD40-binding antibodies
WO2020159368A1 (en) 2019-02-01 2020-08-06 Lava Therapeutics B.V. Novel cd40-binding antibodies
WO2020204708A1 (en) 2019-03-29 2020-10-08 Merus N.V. Cd3 binding molecules
US12460002B2 (en) 2019-03-29 2025-11-04 Merus N.V. CD3 binding molecules
WO2020225456A1 (en) 2019-05-09 2020-11-12 Genmab B.V. Dosage regimens for a combination of anti-dr5 antibodies for use in treating cancer
WO2021001289A1 (en) 2019-07-02 2021-01-07 F. Hoffmann-La Roche Ag Immunoconjugates comprising a mutant interleukin-2 and an anti-cd8 antibody
WO2021006199A1 (ja) 2019-07-05 2021-01-14 小野薬品工業株式会社 Ｐｄ－１／ｃｄ３二重特異性タンパク質による血液がん治療
WO2021009146A1 (en) 2019-07-15 2021-01-21 F. Hoffmann-La Roche Ag Antibodies binding to nkg2d
WO2021020416A1 (ja) 2019-07-30 2021-02-04 小野薬品工業株式会社 二重特異性抗体
WO2021018859A2 (en) 2019-07-31 2021-02-04 F. Hoffmann-La Roche Ag Antibodies binding to gprc5d
WO2021018925A1 (en) 2019-07-31 2021-02-04 F. Hoffmann-La Roche Ag Antibodies binding to gprc5d
WO2021025140A1 (ja) 2019-08-08 2021-02-11 小野薬品工業株式会社 二重特異性タンパク質
WO2021034194A2 (en) 2019-08-19 2021-02-25 Merus N.V. Treatment of cancer with a combination of an antibody that binds lgr5 and egfr and a topoisomerase i inhibitor.
EP3792283A1 (en) 2019-09-16 2021-03-17 Lava Therapeutics B.V. Treatment of cancer comprising administration of vgamma9vdelta2 t cell receptor binding antibodies
WO2021052995A1 (en) 2019-09-16 2021-03-25 Lava Therapeutics B.V. TREATMENT OF CANCER COMPRISING ADMINISTRATION OF Vγ9Vẟ2 T CELL RECEPTOR BINDING ANTIBODIES
WO2021089850A1 (en) 2019-11-06 2021-05-14 Genmab B.V. Antibody variant combinations and uses thereof
US12103967B2 (en) 2019-12-18 2024-10-01 Hoffmann-La Roche Inc. Bispecific anti-CCL2 antibodies
US11987632B2 (en) 2019-12-18 2024-05-21 Hoffmann-La Roche Inc. Antibodies binding to HLA-A2/MAGE-A4
WO2021122733A1 (en) 2019-12-18 2021-06-24 F. Hoffmann-La Roche Ag Bispecific anti-ccl2 antibodies
US11739142B2 (en) 2019-12-18 2023-08-29 Hoffmann-La Roche Inc. Bispecific anti-CCL2 antibodies
WO2021122875A1 (en) 2019-12-18 2021-06-24 F. Hoffmann-La Roche Ag Antibodies binding to hla-a2/mage-a4
WO2021144457A1 (en) 2020-01-16 2021-07-22 Genmab A/S Formulations of cd38 antibodies and uses thereof
WO2021154073A1 (en) 2020-01-29 2021-08-05 Merus N.V. Means and method for modulating immune cell engaging effects.
WO2021156258A1 (en) 2020-02-04 2021-08-12 BioNTech SE Treatment involving antigen vaccination and binding agents binding to pd-l1 and cd137
WO2021155916A1 (en) 2020-02-04 2021-08-12 BioNTech SE Treatment involving antigen vaccination and binding agents binding to pd-l1 and cd137
WO2021185934A1 (en) 2020-03-18 2021-09-23 Genmab A/S Antibodies binding to b7h4
WO2021209402A2 (en) 2020-04-15 2021-10-21 F. Hoffmann-La Roche Ag Immunoconjugates
WO2021215926A1 (en) 2020-04-24 2021-10-28 Merus N.V. Treatment of cancers with an antibody that binds lgr5 and egfr
WO2021222552A1 (en) 2020-04-30 2021-11-04 Celgene Corporation Methods of treating cytokine-related adverse events
US12435154B2 (en) 2020-05-08 2025-10-07 Genmab A/S Bispecific antibodies against CD3 and CD20
AU2021274522B2 (en) * 2020-05-21 2025-10-09 Merus N.V. Methods and means for the production of Ig-like molecules
WO2021235936A1 (en) 2020-05-21 2021-11-25 Merus N.V. Methods and means for the production of ig-like molecules
WO2021255142A1 (en) 2020-06-19 2021-12-23 F. Hoffmann-La Roche Ag Antibodies binding to cd3
WO2021255143A1 (en) 2020-06-19 2021-12-23 F. Hoffmann-La Roche Ag Antibodies binding to cd3 and folr1
US11780920B2 (en) 2020-06-19 2023-10-10 Hoffmann-La Roche Inc. Antibodies binding to CD3 and CD19
WO2021255138A1 (en) 2020-06-19 2021-12-23 F. Hoffmann-La Roche Ag Immune activating fc domain binding molecules
WO2021255155A1 (en) 2020-06-19 2021-12-23 F. Hoffmann-La Roche Ag Antibodies binding to cd3 and cd19
WO2021255146A1 (en) 2020-06-19 2021-12-23 F. Hoffmann-La Roche Ag Antibodies binding to cd3 and cea
US12215155B2 (en) 2020-06-19 2025-02-04 Hoffmann-La Roche Inc. Antibodies binding to CD3 and CD19
WO2022008646A1 (en) 2020-07-08 2022-01-13 LAVA Therapeutics N.V. Antibodies that bind psma and gamma-delta t cell receptors
WO2022018294A1 (en) 2020-07-23 2022-01-27 Genmab B.V. A combination of anti-dr5 antibodies and an immunomodulatory imide drug for use in treating multiple myeloma
WO2022023559A1 (en) 2020-07-31 2022-02-03 Curevac Ag Nucleic acid encoded antibody mixtures
WO2022029011A1 (en) 2020-08-06 2022-02-10 BioNTech SE Binding agents for coronavirus s protein
US11845805B2 (en) 2020-09-10 2023-12-19 Genmab A/S Bispecific antibody against CD3 and CD20 in combination therapy for treating diffuse large B-cell lymphoma
US11858995B2 (en) 2020-09-10 2024-01-02 Genmab A/S Bispecific antibodies against CD3 and CD20 for treating chronic lymphocytic leukemia
WO2022069724A1 (en) 2020-10-02 2022-04-07 Genmab A/S Antibodies capable of binding to ror2 and bispecific antibodies binding to ror2 and cd3
WO2022122973A1 (en) 2020-12-10 2022-06-16 LAVA Therapeutics N.V. Antibodies that bind gamma-delta t cell receptors
WO2022131912A1 (en) 2020-12-15 2022-06-23 Merus N.V. Treatment of cancers with an antibody that binds lgr5 and egfr
WO2022129120A1 (en) 2020-12-17 2022-06-23 F. Hoffmann-La Roche Ag Anti-hla-g antibodies and use thereof
US12466889B2 (en) 2020-12-17 2025-11-11 Hoffmann-La Roche Inc. Anti-HLA-G antibodies and use thereof
WO2022148732A1 (en) 2021-01-06 2022-07-14 F. Hoffmann-La Roche Ag Combination therapy employing a pd1-lag3 bispecific antibody and a cd20 t cell bispecific antibody
WO2022148853A1 (en) 2021-01-11 2022-07-14 F. Hoffmann-La Roche Ag Immunoconjugates
WO2022180271A1 (en) 2021-02-26 2022-09-01 LAVA Therapeutics N.V. Antibodies that bind CD123 and gamma-delta T cell receptors
WO2022211625A1 (en) 2021-03-31 2022-10-06 Merus N.V. Novel multispecific antibodies
WO2022228705A1 (en) 2021-04-30 2022-11-03 F. Hoffmann-La Roche Ag Dosing for combination treatment with anti-cd20/anti-cd3 bispecific antibody and anti-cd79b antibody drug conjugate
US12195547B2 (en) 2021-04-30 2025-01-14 Hoffmann-La Roche Inc. Dosing for combination treatment with anti-CD20/anti-CD3 bispecific antibody and anti-CD79B antibody drug conjugate
WO2022228706A1 (en) 2021-04-30 2022-11-03 F. Hoffmann-La Roche Ag Dosing for treatment with anti-cd20/anti-cd3 bispecific antibody
WO2022234146A1 (en) 2021-05-07 2022-11-10 Genmab A/S PHARMACEUTICAL COMPOSITIONS COMPRISING BISPECIFIC ANTIBODIES BINDING TO B7H4 and CD3
WO2022263501A1 (en) 2021-06-18 2022-12-22 F. Hoffmann-La Roche Ag Bispecific anti-ccl2 antibodies
WO2022268740A1 (en) 2021-06-21 2022-12-29 Genmab A/S Combination dosage regime of cd137 and pd-l1 binding agents
WO2023279092A2 (en) 2021-07-02 2023-01-05 Genentech, Inc. Methods and compositions for treating cancer
WO2023001884A1 (en) 2021-07-22 2023-01-26 F. Hoffmann-La Roche Ag Heterodimeric fc domain antibodies
WO2023010095A1 (en) 2021-07-28 2023-02-02 F. Hoffmann-La Roche Ag Methods and compositions for treating cancer
WO2023031473A1 (en) 2021-09-06 2023-03-09 Genmab B.V. Antibodies capable of binding to cd27, variants thereof and uses thereof
WO2023037333A1 (en) 2021-09-13 2023-03-16 Janssen Biotech, Inc CD33 X Vδ2 MULTISPECIFIC ANTIBODIES FOR THE TREATMENT OF CANCER
WO2023059191A1 (en) 2021-10-06 2023-04-13 Merus N.V. Treatment of immune checkpoint inhibitor-treated cancers with high egfr expression using an antibody that binds at least egfr
WO2023057571A1 (en) 2021-10-08 2023-04-13 Genmab A/S Antibodies binding to cd30 and cd3
WO2023062048A1 (en) 2021-10-14 2023-04-20 F. Hoffmann-La Roche Ag Alternative pd1-il7v immunoconjugates for the treatment of cancer
WO2023062050A1 (en) 2021-10-14 2023-04-20 F. Hoffmann-La Roche Ag New interleukin-7 immunoconjugates
WO2023067138A1 (en) 2021-10-21 2023-04-27 LAVA Therapeutics N.V. Uses of gamma delta t cell activating antibodies
WO2023089083A1 (en) 2021-11-19 2023-05-25 Merus N.V. Multispecific binding moieties comprising pd-1 and tgf-brii binding domains
WO2023094282A1 (en) 2021-11-25 2023-06-01 F. Hoffmann-La Roche Ag Quantification of low amounts of antibody sideproducts
WO2023104938A1 (en) 2021-12-10 2023-06-15 F. Hoffmann-La Roche Ag Antibodies binding to cd3 and plap
WO2023172133A1 (en) 2022-03-07 2023-09-14 Merus N.V. Combination therapy including antibodies that bind egfr and cmet
WO2023172134A1 (en) 2022-03-07 2023-09-14 Merus N.V. Treatment with an antibody that binds egfr and cmet.
WO2023174521A1 (en) 2022-03-15 2023-09-21 Genmab A/S Binding agents binding to epcam and cd137
WO2023174952A1 (en) 2022-03-15 2023-09-21 Genmab A/S Binding agents binding to epcam and/or cd137
WO2023180353A1 (en) 2022-03-23 2023-09-28 F. Hoffmann-La Roche Ag Combination treatment of an anti-cd20/anti-cd3 bispecific antibody and chemotherapy
WO2023198727A1 (en) 2022-04-13 2023-10-19 F. Hoffmann-La Roche Ag Pharmaceutical compositions of anti-cd20/anti-cd3 bispecific antibodies and methods of use
WO2023218046A1 (en) 2022-05-12 2023-11-16 Genmab A/S Binding agents capable of binding to cd27 in combination therapy
WO2023218051A1 (en) 2022-05-12 2023-11-16 Genmab A/S Binding agents capable of binding to cd27 in combination therapy
WO2023232805A1 (en) 2022-05-30 2023-12-07 LAVA Therapeutics N.V. Combination treatment for chronic lymphocytic leukemia
EP4285926A1 (en) 2022-05-30 2023-12-06 LAVA Therapeutics N.V. Combination treatment for chronic lymphocytic leukemia
EP4292609A1 (en) 2022-06-15 2023-12-20 LAVA Therapeutics N.V. Compositions comprising antibodies that bind gamma-delta t cell receptors
EP4292610A1 (en) 2022-06-15 2023-12-20 LAVA Therapeutics N.V. Variant antibodies that bind gamma-delta t cell receptors
WO2023242319A1 (en) 2022-06-15 2023-12-21 LAVA Therapeutics N.V. Variant antibodies that bind gamma-delta t cell receptors
WO2023242320A1 (en) 2022-06-15 2023-12-21 LAVA Therapeutics N.V. Compositions comprising antibodies that bind gamma-delta t cell receptors
NL2032398B1 (en) 2022-07-06 2024-01-23 Academisch Ziekenhuis Leiden Bispecific antibody and uses thereof
WO2024010445A1 (en) 2022-07-06 2024-01-11 ACADEMISCH ZIEKENHUIS LEIDEN (h.o.d.n. LUMC) Bispecific antibody and uses thereof
WO2024079015A1 (en) 2022-10-10 2024-04-18 F. Hoffmann-La Roche Ag Combination therapy of a gprc5d tcb and imids
WO2024079009A1 (en) 2022-10-10 2024-04-18 F. Hoffmann-La Roche Ag Combination therapy of a gprc5d tcb and proteasome inhibitors
WO2024079010A1 (en) 2022-10-10 2024-04-18 F. Hoffmann-La Roche Ag Combination therapy of a gprc5d tcb and cd38 antibodies
WO2024094660A1 (en) 2022-10-31 2024-05-10 Genmab A/S Cd38 antibodies and uses thereof
WO2024100170A1 (en) 2022-11-11 2024-05-16 F. Hoffmann-La Roche Ag Antibodies binding to hla-a*02/foxp3
WO2024104933A1 (en) 2022-11-15 2024-05-23 F. Hoffmann-La Roche Ag Antigen binding molecules
WO2024133330A1 (en) 2022-12-20 2024-06-27 Merus N.V. FAP BINDING DOMAINS AND BISPECIFIC BINDING MOIETIES THAT BIND FAP AND TGF-βRII
WO2024136658A1 (en) 2022-12-23 2024-06-27 Merus N.V. Combination treatment of cancers using an antibody that binds at least egfr and an immune checkpoint inhibitor
WO2024143442A1 (ja) 2022-12-27 2024-07-04 中外製薬株式会社 会合が制御されたポリペプチド
EP4644548A1 (en) 2022-12-27 2025-11-05 Chugai Seiyaku Kabushiki Kaisha Polypeptide with controlled association
WO2024144401A1 (en) 2022-12-28 2024-07-04 Merus N.V. Treatment of cancer with a combination of an antibody that binds egfr and cytotoxic drugs
WO2024153722A1 (en) 2023-01-20 2024-07-25 F. Hoffmann-La Roche Ag Immunoconjugates
WO2024156672A1 (en) 2023-01-25 2024-08-02 F. Hoffmann-La Roche Ag Antibodies binding to csf1r and cd3
WO2024163009A1 (en) 2023-01-31 2024-08-08 Genentech, Inc. Methods and compositions for treating urothelial bladder cancer
WO2024163494A1 (en) 2023-01-31 2024-08-08 F. Hoffmann-La Roche Ag Methods and compositions for treating non-small cell lung cancer and triple-negative breast cancer
WO2024184287A1 (en) 2023-03-06 2024-09-12 F. Hoffmann-La Roche Ag Combination therapy of an anti-egfrviii/anti-cd3 antibody and an tumor-targeted 4-1bb agonist
WO2024188965A1 (en) 2023-03-13 2024-09-19 F. Hoffmann-La Roche Ag Combination therapy employing a pd1-lag3 bispecific antibody and an hla-g t cell bispecific antibody
EP4438624A1 (en) 2023-03-27 2024-10-02 LAVA Therapeutics N.V. Antibodies that bind nectin-4 and gamma-delta t cell receptors
WO2024200573A1 (en) 2023-03-27 2024-10-03 LAVA Therapeutics N.V. Nectin-4 binding agents and methods of use
WO2024208777A1 (en) 2023-04-03 2024-10-10 F. Hoffmann-La Roche Ag All-in-one agonistic antibodies
WO2024208776A1 (en) 2023-04-03 2024-10-10 F. Hoffmann-La Roche Ag Agonistic split antibodies
WO2024208898A1 (en) 2023-04-05 2024-10-10 Genmab A/S Pharmaceutical compositions comprising antibodies binding to cd30 and cd3
WO2024231320A1 (en) 2023-05-08 2024-11-14 F. Hoffmann-La Roche Ag Targeted interferon alpha fusion proteins and methods of use
US12410258B2 (en) 2023-05-12 2025-09-09 Ganmab A/S Antibodies capable of binding to OX40, variants thereof and uses thereof
WO2024235862A1 (en) 2023-05-12 2024-11-21 Genmab A/S Antibodies capable of binding to ox40, variants thereof and uses thereof
WO2024238537A1 (en) 2023-05-16 2024-11-21 F. Hoffmann-La Roche Ag Pd-1 -regulated il-2 immunocytokine and uses thereof
WO2024263904A1 (en) 2023-06-23 2024-12-26 Genentech, Inc. Methods for treatment of liver cancer
WO2024263195A1 (en) 2023-06-23 2024-12-26 Genentech, Inc. Methods for treatment of liver cancer
US12351650B2 (en) 2023-06-30 2025-07-08 Genmab A/S Antibodies binding to fibroblast activation protein alpha and death receptor 4
WO2025003280A1 (en) 2023-06-30 2025-01-02 Genmab A/S Antibodies binding to fibroblast activation protein alpha and death receptor 4
WO2025012118A2 (en) 2023-07-07 2025-01-16 LAVA Therapeutics N.V. 5t4 binding agents and methods of use
WO2025021838A1 (en) 2023-07-26 2025-01-30 F. Hoffmann-La Roche Ag Antibodies binding to cd3
WO2025024780A1 (en) 2023-07-27 2025-01-30 LAVA Therapeutics N.V. Antibodies that bind gamma-delta t cell receptors for the treatment of cancer
WO2025027511A1 (en) 2023-07-30 2025-02-06 Janssen Biotech, Inc. Molecules that bind to mutant calreticulin and uses thereof
WO2025042742A1 (en) 2023-08-18 2025-02-27 Bristol-Myers Squibb Company Compositions comprising antibodies that bind bcma and cd3 and methods of treatment
WO2025056778A1 (en) 2023-09-15 2025-03-20 BioNTech SE Methods of treatment using agents binding to epcam and cd137 in combination with pd-1 axis binding antagonists
WO2025056180A1 (en) 2023-09-15 2025-03-20 BioNTech SE Methods of treatment using agents binding to epcam and cd137 in combination with pd-1 axis binding antagonists
WO2025105961A1 (en) 2023-11-17 2025-05-22 Merus N.V. COMBINATION THERAPY INCLUDING ANTIBODIES THAT BIND EGFR AND cMET.
WO2025114541A1 (en) 2023-11-30 2025-06-05 Genmab A/S Antibodies capable of binding to ox40 in combination therapy
WO2025125386A1 (en) 2023-12-14 2025-06-19 F. Hoffmann-La Roche Ag Antibodies that bind to folr1 and methods of use
WO2025181189A1 (en) 2024-03-01 2025-09-04 F. Hoffmann-La Roche Ag Antibodies binding to cd3
WO2025202147A1 (en) 2024-03-27 2025-10-02 F. Hoffmann-La Roche Ag Interleukin-7 immunoconjugates
WO2025221138A1 (en) 2024-04-15 2025-10-23 Merus N.V. Cd3 binding domains
WO2025219504A1 (en) 2024-04-19 2025-10-23 F. Hoffmann-La Roche Ag Treatment of ophthalmologic diseases
WO2025230409A1 (en) 2024-04-30 2025-11-06 Merus N.V. Multispecific antibody-drug conjugates
WO2025230410A1 (en) 2024-04-30 2025-11-06 Merus N.V. Multispecific antibody-drug conjugates
WO2025233431A1 (en) 2024-05-07 2025-11-13 Immatics Biotechnologies Gmbh Heteromeric proteins comprising three heteromerization improving substitution, production, combinations and applications thereof

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