US20030166868A1 - Polypeptide variants - Google Patents

Polypeptide variants Download PDF

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US20030166868A1
US20030166868A1 US09/792,938 US79293801A US2003166868A1 US 20030166868 A1 US20030166868 A1 US 20030166868A1 US 79293801 A US79293801 A US 79293801A US 2003166868 A1 US2003166868 A1 US 2003166868A1
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region
polypeptide
variant
amino acid
binding
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Leonard Presta
Robert Shields
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Genentech Inc
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Genentech Inc
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
    • A61K47/6873—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting an immunoglobulin; the antibody being an anti-idiotypic antibody
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C07K2317/734—Complement-dependent cytotoxicity [CDC]

Definitions

  • the present invention concerns variants of polypeptides comprising an Fc region. More particularly, the present invention concerns Fc region-containing polypeptides that have altered effector function as a consequence of one or more amino acid substitutions in the Fc region of the nonvariant polypeptide.
  • the invention also relates to novel immune complexes and an assay for determining binding of an analyte, such as an Fc region-containing polypeptide, to a receptor.
  • Antibodies are proteins, which exhibit binding specificity to a specific antigen.
  • Native antibodies are usually heterotetrameric glycoproteins of about 150,000 daltons, composed of two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide linkages varies between the heavy chains of different immunoglobulin isotypes.
  • Each heavy and light chain also has regularly spaced intrachain disulfide bridges.
  • Each heavy chain has at one end a variable domain (V H ) followed by a number of constant domains.
  • V H variable domain
  • Each light chain has a variable domain at one end (V L ) and a constant domain at its other end; the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light chain variable domain is aligned with the variable domain of the heavy chain. Particular amino acid residues are believed to form an interface between the light and heavy chain variable domains.
  • variable refers to the fact that certain portions of the variable domains differ extensively in sequence among antibodies and are responsible for the binding specificity of each particular antibody for its particular antigen. However, the variability is not evenly distributed through the variable domains of antibodies. It is concentrated in three segments called complementarity determining regions (CDRs) both in the light chain and the heavy chain variable domains. The more highly conserved portions of the variable domains are called the framework regions (FR).
  • CDRs complementarity determining regions
  • FR framework regions
  • the variable domains of native heavy and light chains each comprise four FR regions, largely adopting a ⁇ sheet configuration, connected by three CDRs, which form loops connecting, and in some cases forming part of, the ⁇ -sheet structure.
  • the CDRs in each chain are held together in close proximity by the FR regions and, with the CDRs from the other chain, contribute to the formation of the antigen binding site of antibodies (see Kabat et al., Sequences of Proteins of Immunological Interest , 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)).
  • the constant domains are not involved directly in binding an antibody to an antigen, but exhibit various effector functions.
  • antibodies or immunoglobulins can be assigned to different classes.
  • the heavy chain constant regions that correspond to the different classes of immunoglobulins are called ⁇ , ⁇ , ⁇ , ⁇ , and ⁇ , respectively.
  • human immunoglobulin classes only human IgG1, IgG2, IgG3 and IgM are known to activate complement.
  • FIG. 1 A schematic representation of the native IgG1 structure is shown in FIG. 1, where the various portions of the native antibody molecule are indicated.
  • Papain digestion of antibodies produces two identical antigen binding fragments, called Fab fragments, each with a single antigen binding site, and a residual “Fc” fragment, whose name reflects its ability to crystallize readily.
  • the crystal structure of the human IgG Fc region has been determined (Deisenhofer, Biochemistry 20:2361-2370 (1981)).
  • the Fc region is generated by papain cleavage N-terminal to Cys 226.
  • the Fc region is central to the effector functions of antibodies.
  • effector functions mediated by the antibody Fc region can be divided into two categories: (1) effector functions that operate after the binding of antibody to an antigen (these functions involve the participation of the complement cascade or Fc receptor (FcR)-bearing cells); and (2) effector functions that operate independently of antigen binding (these functions confer persistence in the circulation and the ability to be transferred across cellular barriers by transcytosis).
  • effector functions that operate after the binding of antibody to an antigen (these functions involve the participation of the complement cascade or Fc receptor (FcR)-bearing cells); and (2) effector functions that operate independently of antigen binding (these functions confer persistence in the circulation and the ability to be transferred across cellular barriers by transcytosis).
  • FcR complement cascade or Fc receptor
  • an antibody While binding of an antibody to the requisite antigen has a neutralizing effect that might prevent the binding of a foreign antigen to its endogenous target (e.g. receptor or ligand), binding alone may not remove the foreign antigen.
  • an antibody To be efficient in removing and/or destructing foreign antigens, an antibody should be endowed with both high affinity binding to its antigen, and efficient effector functions.
  • C1q and two serine proteases, C1r and C1s form the complex C1, the first component of the complement dependent cytotoxicity (CDC) pathway.
  • C1q is a hexavalent molecule with a molecular weight of approximately 460,000 and a structure likened to a bouquet of tulips in which six collagenous “stalks” are connected to six globular head regions.
  • Burton and Woof Advances in Immunol . 51:1-84 (1992).
  • C1q To activate the complement cascade, it is necessary for C1q to bind to at least two molecules of IgG1, IgG2, or IgG3 (the consensus is that IgG4 does not activate complement), but only one molecule of IgM, attached to the antigenic target. Ward and Ghetie, Therapeutic Immunology 2:77-94 (1995) at page 80.
  • the residue Pro331 has been implicated in C1q binding by analysis of the ability of human IgG subclasses to carry out complement mediated cell lysis. Mutation of Ser331 to Pro331 in IgG4 conferred the ability to activate complement. (Tao et aL, J. Exp. Med ., 178:661-667 (1993); Brekke et al., Eur. J. lmmunol , 24:2542-47 (1994)).
  • Effector functions can also be mediated by the interaction of the Fc region of an antibody with Fc receptors (FcRs), which are specialized cell surface receptors on hematopoietic cells.
  • Fc receptors belong in the immunoglobulin superfamily, and have been shown to mediate both the removal of antibody-coated pathogens by phagocytosis of immune complexes, and the lysing of erythrocytes and various other cellular targets (e.g. tumor cells) coated with the corresponding antibody, via antibody dependent cell mediated cytotoxicity (ADCC). Van de Winkel and Anderson, J. Leuk. Biol . 49:511-24 (1991).
  • FcRs are defined by their specificity for immunoglobulin isotypes; Fc receptors for IgG antibodies are referred to as Fc ⁇ R, for IgE as Fc ⁇ R, for IgA as Fc ⁇ R and so on.
  • Fc ⁇ RI CD64
  • Fc ⁇ RII CD32
  • Fc ⁇ RIII CD16
  • the three genes encoding the Fc ⁇ RI subclass are clustered in region 1q21.1 of the long arm of chromosome 1; the genes encoding Fc ⁇ RII isoforms (Fc ⁇ RIIA, Fc ⁇ RIIB and Fc ⁇ RIIC) and the two genes encoding Fc ⁇ RIII (Fc ⁇ RIIIA and Fc ⁇ RIIIB) are all clustered in region 1q22.
  • FcRs are reviewed in Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991); Capel et aL, Immunomethods 4:25-34 (1994); and de Haas etal., J. Lab. Clin. Med . 126:330-41 (1995).
  • Fc ⁇ RI binds monomeric IgG with a high affinity
  • Fc ⁇ RII and Fc ⁇ RIII are low-affinity receptors, interacting with complexed or aggregated IgG.
  • the classical method for detecting these low-affinity receptors is by “rosetting” using antibody-coated erythrocytes (EA) sensitized with IgGs.
  • EA antibody-coated erythrocytes
  • Bredius et aL evaluated rosette formation between IgG-sensitized red blood cells and polymorphonuclear leukocytes (PMN) which express Fc ⁇ RIIa and Fc ⁇ RIIIb at their cell-surface. Rosette was defined as three of more EA bound per PMN (Bredius etal.
  • IgG dimers have been formed using anti-light chain monoclonal antibodies (Nagarajan et al, supra and Warmerdam et al., supra) or chemical cross-linking agents (Hogarth et al. Immunomethods 4:17-24 (1994); and Tamm et aL J. Biol Chem . 271(7):3659-3666(1996)). Heat-aggregated immune complexes have also been evaluated for binding to cells expressing FcRs (Tax et al., supra and Tam et al., supra).
  • Pro331 in IgG3 was changed to Ser, and the affinity of this mutant to target cells analyzed. The affinity was found to be six-fold lower than that of unmutated IgG3, indicating the involvement of Pro331 in Fc ⁇ RI binding. Morrison et al., Immunologist , 2:119-124 (1994); and Canfield and Morrison, J. Exp. Med . 173:1483-91 (1991).
  • Glu318 was identified as being involved in binding to Fc ⁇ RII. Ward and Ghetie, Therapeutic Immunology 2:77-94 (1995).
  • the present invention provides a variant of a polypeptide comprising a human IgG Fc region, which variant comprises an amino acid substitution at amino acid position 329, or at two or all of amino acid positions 329, 331 and 322 of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat.
  • the invention further relates to a variant of a polypeptide comprising a human IgG Fc region, which variant binds Fc ⁇ RI, Fc ⁇ RII, Fc ⁇ RIII and FcRn but does not activate complement and comprises an amino acid substitution at amino acid position 322 or amino acid position 329, or both amino acid positions of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat.
  • the invention provides a method for modifying a polypeptide comprising a human IgG Fc region comprising substituting an amino acid residue at amino acid position 329, or at two or all of amino acid positions 329, 331 and 322 of the human lgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat.
  • the invention also provides a composition comprising the polypeptide variant and a physiologically acceptable carrier or diluent.
  • This composition for potential therapeutic use is sterile and may be lyophilized.
  • the invention provides a method for determining the presence of a protein of interest comprising exposing a sample suspected of containing the protein to the polypeptide variant and determining binding of the polypeptide variant to the sample.
  • the invention provides a method of treating a mammal suffering from a disorder comprising administering to the mammal a therapeutically effective amount of a variant of a polypeptide comprising a human IgG Fc region, which variant binds Fc ⁇ RI, Fc ⁇ RII, Fc ⁇ RIII and FcRn but does not activate complement and comprises an amino acid substitution at amino acid position 329, 331 or 322 of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat.
  • the invention further provides: isolated nucleic acid encoding the polypeptide variant; a vector comprising the nucleic acid, optionally, operably linked to control sequences recognized by a host cell transformed with the vector; a host cell comprising the vector; a process for producing the polypeptide variant comprising culturing this host cell so that the nucleic acid is expressed and, optionally, recovering the polypeptide variant from the host cell culture (e.g. from the host cell culture medium).
  • the invention also pertains to an immune complex comprising: (a) an Fc region-containing polypeptide; (b) a first target molecule which comprises at least two binding sites for the Fc region-containing polypeptide; and (c) a second target molecule comprises at least two binding sites for the first target molecule.
  • the immune complex may be used in an FcR-binding assay, particularly where the FcR has a low affinity for the Fc region-containing polypeptide. Other uses for the immune complex are disclosed herein.
  • the invention provides a method for determining binding of an analyte, such as an Fc region-containing polypeptide, to a receptor (e.g. a low affinity FcR) comprising the following steps performed sequentially: (a) forming a molecular complex between the analyte and a first target molecule, wherein the first target molecule comprises at least two binding sites for the analyte; and (b) determining binding of the molecular complex of step (a) to the receptor (e.g. to a binding domain of the receptor coated on an assay plate).
  • the molecular complex of step (a) further comprises a second target molecule which comprises at least two binding sites for the first target molecule.
  • the invention also relates to an assay kit, such as a kit useful for determining binding of an analyte to a receptor comprising: (a) a first target molecule which comprises at least two binding sites for the analyte; and (b) a second target molecule which comprises at least two binding sites for the first target molecule.
  • FIG. 1 is a schematic representation of a native IgG. Disulfide bonds are represented by heavy lines between CH1 and CL domains and the two CH2 domains. V is variable domain; C is constant domain; L stands for light chain and H stands for heavy chain.
  • FIG. 2 shows C1q binding of wild type (wt) C2B8 antibody; C2B8 antibody with a human IgG2 constant region (IgG2); and mutants K322A, K320A and E318A.
  • FIG. 3 depicts C1q binding of mutants P331A, P329A and K322A.
  • FIGS. 4A and 4B depict the amino acid sequences of E27 anti-IgE antibody light chain (FIG. 4A) and heavy chain (FIG. 4B).
  • FIG. 5 is a schematic diagram of the “immune complex” prepared for use in the FcR assay described in Example 1.
  • the hexamer comprising three anti-IgE antibody molecules (the “Fc region-containing polypeptide”) and three IgE molecules (the “first target molecule”) is shown.
  • IgE has two “binding sites” for the anti-lgE antibody (E27) in the Fc region thereof.
  • Each IgE molecule in the complex is further able to bind two VEGF molecules (“the second target polypeptide”).
  • VEGF has two “binding sites” for IgE.
  • the numbering of the residues in an IgG heavy chain is that of the EU index as in Kabat et al., Sequences of Proteins of Immunological Interest , 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991), expressly incorporated herein by reference.
  • the “EU index as in Kabat” refers to the residue numbering of the human IgG1 EU antibody.
  • Fc region is used to define a C-terminal region of an IgG heavy chain as shown FIG. 1. Although the boundaries of the Fc region of an IgG heavy chain might vary slightly, the human IgG heavy chain Fc region is usually defined to stretch from amino acid residue at position Cys226 to the carboxyl-terminus.
  • Fc region-containing polypeptide refers to a polypeptide, such as an antibody or immunoadhesin (see definitions below), which comprises an Fc region.
  • the Fc region of an IgG comprises two constant domains, CH2 and CH3, as shown in FIG. 1.
  • the “CH2” domain of a human IgG Fc region (also referred to as “C ⁇ 2” domain) usually extends from amino acid 231 to amino acid 340.
  • the CH2 domain is unique in that it is not closely paired with another domain. Rather, two N-linked branched carbohydrate chains are interposed between the two CH2 domains of an intact native IgG molecule. It has been speculated that the carbohydrate may provide a substitute for the domain-domain pairing and help stabilize the CH2 domain.
  • Hinge region is generally defined as stretching from Glu216 to Pro230 of human IgG1 (Burton, Molec. Immunol . 22:161-206 (1985)) Hinge regions of other IgG isotypes may be aligned with the IgG1 sequence by placing the first and last cysteine residues forming inter-heavy chain S-S bonds in the same positions.
  • C1q is a polypeptide that includes a binding site for the Fc region of an immunoglobulin. C1q together with two serine proteases, C1r and C1s, forms the complex C1, the first component of the complement dependent cytotoxicity (CDC) pathway. Human C1q can be purchased commercially from, e.g. Quidel, San Diego, CA.
  • Fc receptor or “FcR” is used to describe a receptor that binds to the Fc region of an antibody.
  • the preferred FcR is one, which binds an IgG antibody (a gamma receptor) and includes receptors of the Fc ⁇ RI, Fc ⁇ RII, and Fc ⁇ RIII subclasses, including allelic variants and alternatively spliced forms of these receptors.
  • FcRs are reviewed in Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991); Capel et al, Immunomethods 4:25-34 (1994); and de Haas et al, J. Lab. Clin. Med . 126:330-41 (1995).
  • FcR FcR
  • FcRn neonatal receptor
  • binding domain refers to the region of a polypeptide that binds to another molecule.
  • the binding domain can comprise a portion of a polypeptide chain thereof (e.g. the ⁇ chain thereof) which is responsible for binding an Fc region.
  • One useful binding domain is the extracellular domain of an FcR ⁇ chain.
  • antibody is used in the broadest sense and specifically covers monoclonal antibodies (including full length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments so long as they exhibit the desired biological activity.
  • Antibody fragments comprise a portion of an intact antibody, generally including the antigen binding or variable region of the intact antibody, so long as they retain at least the CH2 region of an IgG immunoglobulin heavy chain constant domain, comprising amino acid residues 322, 329 and 331, and have the ability, alone or in combination with another antibody fragment, to specifically bind a selected antigen.
  • Examples of antibody fragments include linear antibodies; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments.
  • the antibody fragments preferably retain at least part of the hinge and optionally the CH1 region of an IgG heavy chain. More preferably, the antibody fragments retain the entire constant region of an IgG heavy chain, and include an IgG light chain.
  • the term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to conventional (polyclonal) antibody preparations that typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen.
  • the modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method.
  • the monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method first described by Kohler et al, Nature 256:495 (1975), or may be made by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567).
  • the “monoclonal antibodies” may also be isolated from phage antibody libraries using the techniques described in Clackson et al., Nature 352:624-628 (1991) and Marks et al., J. Mol. Biol . 222:581-597 (1991), for example.
  • the monoclonal antibodies herein specifically include “chimeric” antibodies (immunoglobulins) in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (U.S. Pat. No. 4,816,567; and Morrison et al, Proc. Natl. Acad. Sci. USA 81:6851-6855 (1984)).
  • chimeric antibodies immunoglobulins in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in
  • “Humanized” forms of non-human (e.g., murine) antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulin.
  • humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a hypervariable region of the recipient are replaced by residues from a hypervariable region of a non-human species (donor antibody) such as mouse, rat, rabbit or nonhuman primate having the desired specificity, affinity, and capacity.
  • donor antibody such as mouse, rat, rabbit or nonhuman primate having the desired specificity, affinity, and capacity.
  • Fv framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues.
  • humanized antibodies may comprise residues that are not found in the recipient antibody or in the donor antibody. These modifications are made to further refine antibody performance.
  • the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin sequence.
  • the humanized antibody optionally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin.
  • Fc immunoglobulin constant region
  • hypervariable region when used herein refers to the amino acid residues of an antibody which are responsible for antigen-binding.
  • the hypervariable region comprises amino acid residues from a “complementarity determining region” or “CDR” (i.e. residues 24-34 (L1), 50-56 (L2) and 89-97 (L3) in the light chain variable domain and 31-35 (H1), 50-65 (H2) and 95-102 (H3) in the heavy chain variable domain; Kabat et al., Sequences of Proteins of Immunological Interest , 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md.
  • CDR complementarity determining region
  • residues from a “hypervariable loop” i.e. residues 26-32 (L1), 50-52 (L2) and 91-96 (L3) in the light chain variable domain and 26-32 (H1), 53-55 (H2) and 96-101 (H3) in the heavy chain variable domain; Chothia and Lesk J. Mol. Biol . 196:901-917 (1987)).
  • “Framework” or “FR” residues are those variable domain residues other than the hypervariable region residues as herein defined.
  • the term “immunoadhesin” designates antibody-like molecules which combine the “binding domains” of a heterologous “adhesin” protein (e.g. a receptor, ligand or enzyme) with an immunoglobulin constant domain.
  • adhesin e.g. a receptor, ligand or enzyme
  • the immunoadhesins comprise a fusion of the adhesin amino acid sequence with the desired binding specificity which is other than the antigen recognition and binding site (antigen combining site) of an antibody (i.e. is “heterologous”) and an immunoglobulin constant domain sequence.
  • ligand binding domain refers to any native cell-surface receptor or any region or derivative thereof retaining at least a qualitative ligand binding ability of a corresponding native receptor.
  • the receptor is from a cell-surface polypeptide having an extracellular domain that is homologous to a member of the immunoglobulin supergenefamily.
  • receptors which are not members of the immunoglobulin supergenefamily but are nonetheless specifically covered by this definition, are receptors for cytokines, and in particular receptors with tyrosine kinase activity (receptor tyrosine kinases), members of the hematopoietin and nerve growth factor receptor superfamilies, and cell adhesion molecules, e.g. (E-, L- and P-) selectins.
  • receptor binding domain is used to designate any native ligand for a receptor, including cell adhesion molecules, or any region or derivative of such native ligand retaining at least a qualitative receptor binding ability of a corresponding native ligand. This definition, among others, specifically includes binding sequences from ligands for the above-mentioned receptors.
  • an “antibody-immunoadhesin chimera” comprises a molecule that combines at least one binding domain of an antibody (as herein defined) with at least one immunoadhesin (as defined in this application).
  • Exemplary antibody-immunoadhesin chimeras are the bispecific CD4-lgG chimeras described in Berg et al, PNAS ( USA ) 88:4723-4727 (1991) and Chamow et al, J. Immunol . 153:4268 (1994).
  • An “isolated” polypeptide is one that has been identified and separated and/or recovered from a component of its natural environment. Contaminant components of its natural environment are materials that would interfere with diagnostic or therapeutic uses for the polypeptide, and may include enzymes, hormones, and other proteinaceous or nonproteinaceous solutes.
  • the polypeptide will be purified (1) to greater than 95% by weight of polypeptide as determined by the Lowry method, and most preferably more than 99% by weight, (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence by use of a spinning cup sequenator, or (3) to homogeneity by SDS-PAGE under reducing or nonreducing conditions using Coomassie blue or, preferably, silver stain.
  • Isolated polypeptide includes the polypeptide in situ within recombinant cells since at least one component of the polypeptide's natural environment will not be present. Ordinarily, however, isolated polypeptide will be prepared by at least one purification step.
  • Treatment refers to both therapeutic treatment and prophylactic or preventative measures. Those in need of treatment include those already with the disorder as well as those in which the disorder is to be prevented.
  • a “disorder” is any condition that would benefit from treatment with the polypeptide variant. This includes chronic and acute disorders or diseases including those pathological conditions which predispose the mammal to the disorder in question.
  • label when used herein refers to a detectable compound or composition which is conjugated directly or indirectly to the polypeptide.
  • the label may be itself be detectable (e.g., radioisotope labels or fluorescent labels) or, in the case of an enzymatic label, may catalyze chemical alteration of a substrate compound or composition which is detectable.
  • An “isolated” nucleic acid molecule is a nucleic acid molecule that is identified and separated from at least one contaminant nucleic acid molecule with which it is ordinarily associated in the natural source of the polypeptide nucleic acid.
  • An isolated nucleic acid molecule is other than in the form or setting in which it is found in nature. Isolated nucleic acid molecules therefore are distinguished from the nucleic acid molecule as it exists in natural cells.
  • an isolated nucleic acid molecule includes a nucleic acid molecule contained in cells that ordinarily express the polypeptide where, for example, the nucleic acid molecule is in a chromosomal location different from that of natural cells.
  • control sequences refers to DNA sequences necessary for the expression of an operably linked coding sequence in a particular host organism.
  • the control sequences that are suitable for prokaryotes include a promoter, optionally an operator sequence, and a ribosome binding site.
  • Eukaryotic cells are known to utilize promoters, polyadenylation signals, and enhancers.
  • Nucleic acid is “operably linked” when it is placed into a functional relationship with another nucleic acid sequence.
  • DNA for a presequence or secretory leader is operably linked to DNA for a polypeptide if it is expressed as a preprotein that participates in the secretion of the polypeptide;
  • a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence; or
  • a ribosome binding site is operably linked to a coding sequence if it is positioned so as to facilitate translation.
  • “operably linked” means that the DNA sequences being linked are contiguous, and, in the case of a secretory leader, contiguous and in reading phase. However, enhancers do not have to be contiguous. Linking is accomplished by ligation at convenient restriction sites. If such sites do not exist, the synthetic oligonucleotide adaptors or linkers are used in accordance with conventional practice.
  • the expressions “cell,” “cell line,” and “cell culture” are used interchangeably and all such designations include progeny.
  • the words “transformants” and “transformed cells” include the primary subject cell and cultures derived therefrom without regard for the number of transfers. It is also understood that all progeny may not be precisely identical in DNA content, due to deliberate or inadvertent mutations. Mutant progeny that have the same function or biological activity as screened for in the originally transformed cell are included. Where distinct designations are intended, it will be clear from the context.
  • molecular complex when used herein refers to the relatively stable structure which forms when two or more heterologous molecules (e.g. polypeptides) bind (preferably noncovalently) to one another.
  • the preferred molecular complex herein is an immune complex.
  • Immuno complex refers to the relatively stable structure which forms when at least one target molecule and at least one heterologous Fc region-containing polypeptide bind to one another forming a larger molecular weight complex.
  • immune complexes are antigen-antibody aggregates and target molecule-immunoadhesin aggregates.
  • immuno complex as used herein, unless indicated otherwise, refers to an ex vivo complex (i.e. other than the form or setting in which it may be found in nature). However, the immune complex may be administered to a mammal, e.g. to evaluate clearance of the immune complex in the mammal.
  • target molecule refers to a molecule, usually a polypeptide, which is capable of being bound by a heterologous molecule and has one or more binding sites for the heterologous molecule.
  • binding site refers to a region of a molecule to which another molecule can bind.
  • the “first target molecule” herein comprises at least two distinct binding sites (for example, two to five separate binding sites) for an analyte (e.g. an Fc region-containing polypeptide) such that at least two analyte molecules can bind to the first target molecule.
  • the two or more binding sites are identical (e.g. having the same amino acid sequence, where the target molecule is a polypeptide).
  • the first target molecule was IgE and had two separate binding sites in the Fc region thereof to which the Fc region-containing polypeptide (an anti-IgE antibody, E27) could bind.
  • Other first target molecules include dimers of substantially identical monomors (e.g. neurotrophins, IL8 and VEGF) or are polypeptides comprising two or more substantially identical polypeptide chains (e.g. antibodies or immunoadhesins).
  • the “second target molecule” comprises at least two distinct binding sites (for example, two to five separate binding sites) for the first target molecule such that at least two first target molecules can bind to the second target molecule.
  • the two or more binding sites are identical (e.g.
  • the second target molecule was VEGF, which has a pair of distinct binding sites to which the variable domain of the IgE antibody could bind.
  • Other second target molecules are contemplated, e.g. other dimers of substantially identical monomers (e.g. neurotrophins or IL8) or polypeptides comprising two or more substantially identical domains (e.g. antibodies or immunoadhesins).
  • an “analyte” is a substance that is to be analyzed.
  • the preferred analyte is an Fc region-containing polypeptide that is to be analyzed for its ability to bind to an Fc receptor.
  • a “receptor” is a polypeptide capable of binding at least one ligand.
  • the preferred receptor is a cell-surface receptor having an extracellular ligand-binding domain and, optionally, other domains (e.g. transmembrane domain, intracellular domain and/or membrane anchor).
  • the receptor to be evaluated in the assay described herein may be an intact receptor or a fragment or derivative thereof (e.g. a fusion protein comprising the binding domain of the receptor fused to one or more heterologous polypeptides).
  • the receptor to be evaluated for its binding properties may be present in a cell or isolated and optionally coated on an assay plate or some other solid phase.
  • low affinity receptor denotes a receptor that has a weak binding affinity for a ligand of interest, e.g. having a binding constant of about 50 nM or worse affinity.
  • exemplary low affinity receptors include Fc ⁇ RII and Fc ⁇ RIII as well as adhesion molecules, such as integrins.
  • the invention herein relates to a method for making a polypeptide variant.
  • the “starting” or “nonvariant” polypeptide is prepared using techniques available in the art for generating polypeptides comprising an Fc region.
  • the polypeptide is an antibody and exemplary methods for generating antibodies are described in more detail in the following sections.
  • the polypeptide may, however, be any other polypeptide comprising an Fc region, e.g. an immunoadhesin. Methods for making immunoadhesins are elaborated in more detail hereinbelow.
  • the starting polypeptide of particular interest herein is usually one that binds to C1q and displays complement dependent cytotoxicity (CDC).
  • CDC complement dependent cytotoxicity
  • the amino acid substitutions described herein will generally serve to alter the ability of the starting polypeptide to bind to C1q and/or modify its complement dependent cytotoxicity function, e.g. to reduce and preferably abolish these effector functions.
  • polypeptides comprising substitutions at one or more of the described positions with improved effector functions are contemplated herein.
  • the starting polypeptide may be unable to bind C1q and/or mediate CDC and may be modified according to the teachings herein such that it acquires these effector functions.
  • polypeptides with pre-existing C1q binding activity, optionally further having the ability to mediate CDC may be modified such that one or both of these activities are enhanced.
  • one or more amino acid alterations are introduced in the Fc region of the starting polypeptide.
  • the amino acid positions to be modified are selected from heavy chain positions 329, 331 and 322, where the numbering of the residues in an IgG heavy chain is that of the EU index as in Kabat et al, Sequences of Proteins of Immunological Interest , 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md (1991).
  • the Fc region is preferably from a human IgG and most preferably a human IgG1.
  • the human IgG1 Fc region may be a human A or non-A allotype.
  • Proline is conserved at position 329 in human IgG's. This residue is preferably replaced with alanine, however substitution with any other amino acid is contemplated, e.g., serine, threonine, asparagine, glycine or valine. Proline is conserved at position 331 in human IgG1, IgG2 and IgG3, but not IgG4 (which has a serine residue at position 331). Residue 331 is preferably replaced by alanine or another amino acid, e.g. serine (for IgG regions other than IgG4), glycine or valine.
  • Lysine 322 is conserved in human IgGs, and this residue is preferably replaced by an alanine residue, but substitution with any other amino acid residue is contemplated, e.g. serine, threonine, glycine or valine.
  • only one of the three above-identified positions is altered in order to generate the polypeptide variant.
  • residue 329 or only residue 322 is altered if this is the case.
  • two or all of the three identified positions are modified.
  • a variant may be generated in which the native amino acid residue at position 329 of the human heavy chain constant region is substituted with another amino acid, optionally in combination with a substitution of the amino acid residue at position 331 and/or substitution of the amino acid residue at position 322.
  • the native amino acid residue at position 331 and the native amino acid residue at position 322 of the human IgG Fc region may both be substituted with another amino acid residue.
  • DNA encoding amino acid sequence variant of the starting polypeptide is prepared by a variety of methods known in the art. These methods include, but are not limited to, preparation by site-directed (or oligonucleotide-mediated) mutagenesis, PCR mutagenesis, and cassette mutagenesis of an earlier prepared DNA encoding the polypeptide
  • Site-directed mutagenesis is a preferred method for preparing substitution variants. This technique is well known in the art (see, e.g.,Carter et al. Nucleic Acids Res . 13:4431-4443 (1985) and Kunkel et al, Proc. Natl. Acad.Sci.USA 82:488 (1987)). Briefly, in carrying out site-directed mutagenesis of DNA, the starting DNA is altered by first hybridizing an oligonucleotide encoding the desired mutation to a single strand of such starting DNA.
  • a DNA polymerase is used to synthesize an entire second strand, using the hybridized oligonucleotide as a primer, and using the single strand of the starting DNA as a template.
  • the oligonucleotide encoding the desired mutation is incorporated in the resulting double-stranded DNA.
  • PCR mutagenesis is also suitable for making amino acid sequence variants of the starting polypeptide. See Higuchi, in PCR Protocols, pp . 177-183 (Academic Press, 1990); and Vallette et al., Nuc. Acids Res . 17:723-733 (1989). Briefly, when small amounts of template DNA are used as starting material in a PCR, primers that differ slightly in sequence from the corresponding region in a template DNA can be used to generate relatively large quantities of a specific DNA fragment that differs from the template sequence only at the positions where the primers differ from the template.
  • the starting material is the plasmid (or other vector) comprising the starting polypeptide DNA to be mutated.
  • the codon(s) in the starting DNA to be mutated are identified. There must be a unique restriction endonuclease site on each side of the identified mutation site(s). If no such restriction sites exist, they may be generated using the above-described oligonucleotide-mediated mutagenesis method to introduce them at appropriate locations in the starting polypeptide DNA.
  • the plasmid DNA is cut at these sites to linearize it.
  • a double-stranded oligonucleotide encoding the sequence of the DNA between the restriction sites but containing the desired mutation(s) is synthesized using standard procedures, wherein the two strands of the oligonucleotide are synthesized separately and then hybridized together using standard techniques.
  • This double-stranded oligonucleotide is referred to as the cassette.
  • This cassette is designed to have 5′ and 3′ ends that are compatible with the ends of the linearized plasmid, such that it can be directly ligated to the plasmid.
  • This plasmid now contains the mutated DNA sequence.
  • polypeptide variant(s) so prepared may be subjected to further modifications, oftentimes depending on the intended use of the polypeptide. Such modifications may involve further alteration of the amino acid sequence, fusion to heterologous polypeptide(s) and/or covalent modifications. For example, it may be useful to combine the above amino acid substitutions with one or more further amino acid substitutions that reduce or ablate FcR binding.
  • the native amino acid residues at any one or more of heavy chain positions 233-238, 318 or 331 may be replaced with non-native residue(s), e.g. alanine.
  • any cysteine residue not involved in maintaining the proper conformation of the polypeptide variant also may be substituted, generally with serine, to improve the oxidative stability of the molecule and prevent aberrant cross linking.
  • Another type of amino acid substitution serves to alter the glycosylation pattern of the polypeptide. This may be achieved by deleting one or more carbohydrate moieties found in the polypeptide, and/or adding one or more glycosylation sites that are not present in the polypeptide.
  • Glycosylation of polypeptides is typically either N-linked or O-linked. N-linked refers to the attachment of the carbohydrate moiety to the side chain of an asparagine residue.
  • the tripeptide sequences asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline, are the recognition sequences for enzymatic attachment of the carbohydrate moiety to the asparagine side chain.
  • O-linked glycosylation refers to the attachment of one of the sugars N-aceylgalactosamine, galactose, or xylose to a hydroxyamino acid, most commonly serine or threonine, although 5-hydroxyproline or 5-hydroxylysine may also be used.
  • Addition of glycosylation sites to the polypeptide is conveniently accomplished by altering the amino acid sequence such that it contains one or more of the above-described tripeptide sequences (for N-linked glycosylation sites).
  • the alteration may also be made by the addition of, or substitution by, one or more serine or threonine residues to the sequence of the original polypeptide (for O-linked glycosylation sites).
  • An exemplary glycosylation variant has an amino acid substitution of residue Asn 297 of the heavy chain.
  • the polypeptide variant may be subjected to one or more assays to evaluate any change in biological activity compared to the starting polypeptide. For example, the ability of the variant to bind C1q and mediate complement dependent cytotoxicity (CDC) may be assessed.
  • CDC mediate complement dependent cytotoxicity
  • a C1q binding ELISA may be performed. Briefly, assay plates may be coated overnight at 4° C. with polypeptide variant or starting polypeptide (control) in coating buffer. The plates may then be washed and blocked. Following washing, an aliquot of human C1q may be added to each well and incubated for 2 hrs at room temperature. Following a further wash, 100 ⁇ l of a sheep anti-complement C1q peroxidase conjugated antibody may be added to each well and incubated for 1 hour at room temperature. The plate may again be washed with wash buffer and 100 ⁇ l of substrate buffer containing OPD ( O-phenylenediamine dihydrochloride (Sigma)) may be added to each well. The oxidation reaction, observed by the appearance of a yellow color, may be allowed to proceed for 30 minutes and stopped by the addition of 100 ⁇ l of 4.5 N H 2 SO 4 . The absorbance may then read at (492-405) nm.
  • OPD O-phenylenediamine dihydr
  • the preferred polypeptide variant is one that displays a “significant reduction in C1q binding” in this assay. This means that about 100 ⁇ g/ml of the polypeptide variant displays about 50 fold or more reduction in C1q binding compared to 100 ⁇ g/ml of a control antibody having a nonmutated IgG1 Fc region. In the most preferred embodiment, the polypeptide variant “does not bind C1q”, i.e. 100 ⁇ g/ml of the polypeptide variant displays about 100 fold or more reduction in C1q binding compared to 100 ⁇ g/ml of the control antibody.
  • a complement dependent cytotoxicity (CDC) assay may be performed, e.g. as described in Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996). Briefly, various concentrations of the polypeptide variant and human complement may be diluted with buffer. Cells which express the antigen to which the polypeptide variant binds may be diluted to a density of ⁇ 1 ⁇ 10 6 cells/ ml. Mixtures of polypeptide variant, diluted human complement and cells expressing the antigen may be added to a flat bottom tissue culture 96 well plate and allowed to incubate for 2 hrs at 37°C. and 5% CO 2 to facilitate complement mediated cell lysis.
  • CDC complement dependent cytotoxicity
  • alamar blue (Accumed International) may then be added to each well and incubated overnight at 37°C.
  • the absorbance is measured using a 96-well fluorometer with excitation at 530 nm and emission at 590 nm.
  • the results may be expressed in relative fluorescence units (RFU).
  • the sample concentrations may be computed from a standard curve and the percent activity as compared to nonvariant polypeptide is reported for the polypeptide variant of interest.
  • the preferred variant “does not activate complement”.
  • 0.6 ⁇ /ml of the polypeptide variant displays about 0-10% CDC activity in this assay compared to a 0.6 ⁇ /ml of a control antibody having a nonmutated IgG1 Fc region.
  • the variant does not appear to have any CDC activity in the above CDC assay.
  • the polypeptide variant essentially retains the ability to bind antigen compared to the nonvariant polypeptide, i.e. the binding capability is no worse than about 20 fold, e.g. no worse than about 5 fold of that of the nonvariant polypeptide.
  • the binding capability of the polypeptide variant may be determined using techniques such as fluorescence activated cell sorting (FACS) analysis or radioimmunoprecipitation (RIA), for example.
  • the ability of the polypeptide variant to bind an FcR may also be evaluated.
  • the FcR is a high affinity Fc receptor, such as Fc ⁇ RI or FcRn
  • binding can be measured by titrating monomeric polypeptide variant and measuring bound polypeptide variant using an antibody which specifically binds to the polypeptide variant in a standard ELISA format (see Example 2 below).
  • Another FcR binding assay for low affinity FcRs is elaborated in more detail in the following section.
  • the variant retains the ability to bind one or more FcRs, e.g. the ability of the polypeptide variant to bind Fc ⁇ RI, Fc ⁇ RII, Fc ⁇ RIII and/or FcRn is no more than about 20 fold reduced, preferably no more than about 10 fold reduced, and most preferably no more than about two fold reduced compared to the starting polypeptide as determined in the Fc ⁇ RI or FcRn assays of Example 2 or the Fc ⁇ RII or Fc ⁇ RIII assays described in the following section.
  • FcRs e.g. the ability of the polypeptide variant to bind Fc ⁇ RI, Fc ⁇ RII, Fc ⁇ RIII and/or FcRn is no more than about 20 fold reduced, preferably no more than about 10 fold reduced, and most preferably no more than about two fold reduced compared to the starting polypeptide as determined in the Fc ⁇ RI or FcRn assays of Example 2 or the Fc ⁇ RII or Fc ⁇ RIII assays described
  • a receptor binding assay has been developed herein which is particularly useful for determining binding of an analyte of interest to a receptor where the affinity of the analyte for the receptor is relatively weak, e.g. in the micromolar range as is the case for Fc ⁇ RIIa, Fc ⁇ RIIb, Fc ⁇ RIIIa and Fc ⁇ RIIIb.
  • the method involves the formation of a molecular complex that has an improved avidity for the receptor of interest compared to the noncomplexed analyte.
  • the preferred molecular complex is an immune complex comprising: (a) an Fc region-containing polypeptide (such as an antibody or an immunoadhesin); (b) a first target molecule which comprises at least two binding sites for the Fc region-containing polypeptide; and (c) a second target molecule which comprises at least two binding sites for the first target molecule.
  • an Fc region-containing polypeptide such as an antibody or an immunoadhesin
  • a first target molecule which comprises at least two binding sites for the Fc region-containing polypeptide
  • a second target molecule which comprises at least two binding sites for the first target molecule.
  • the Fc region-containing polypeptide is an anti-IgE antibody, such as the E27 antibody (FIGS. 4 A- 4 B).
  • E27 when mixed with human IgE at an 1:1 molar ratio, forms a stable hexamer consisting of three E27 molecules and three IgE molecules.
  • the “first target molecule” is a chimeric form of IgE in which the Fab portion of an anti-VEGF antibody is fused to the human IgE Fc portion and the “second target molecule” is the antigen to which the Fab binds (i.e. VEGF). Each molecule of IgE binds two molecules of VEGF.
  • VEGF also binds two molecules of IgE per molecule of VEGF.
  • recombinant human VEGF was added at a 2:1 molar ratio to IgE:E27 hexamers, the hexamers were linked into larger molecular weight complexes via the IgE:VEGF interaction (FIG. 5).
  • the Fc region of the anti-IgE antibody of the resultant immune complex binds to FcR with higher avidity than either uncomplexed anti-IgE or anti-IgE:lgE hexamers.
  • Fc region-containing polypeptide:first target molecule combination examples include an immunoadhesin:ligand combination such as VEGF receptor (KDR)-immunoadhesin: VEGF and a full-length bispecific antibody (bsAb):first target molecule.
  • an Fc region-containing polypeptide:first target molecule:second target molecule combination include a nonblocking antibody:soluble receptor:ligand combination such as anti-Trk antibody:soluble Trk receptor:neurotrophin (Urfer et al. J. Biol Chem . 273(10):5829-5840 (1998)).
  • the immune complexes described above have further uses including evaluation of Fc region-containing polypeptide function and immune complex clearance in vivo.
  • the immune complex may be administered to a mammal (e.g. in a pre-clinical animal study) and evaluated for its half-life etc.
  • a polypeptide comprising at least the binding domain of the receptor of interest may be coated on solid phase, such as an assay plate.
  • the binding domain of the receptor alone or a receptor-fusion protein may be coated on the plate using standard procedures.
  • receptor-fusion proteins include receptor-glutathione S-transferase (GST) fusion protein, receptor-chitin binding domain fusion protein, receptor-hexaHis tag fusion protein (coated on glutathione, chitin, and nickel coated plates, respectively).
  • a capture molecule may be coated on the assay plate and used to bind the receptor-fusion protein via the non-receptor portion of the fusion protein.
  • examples include anti-hexaHis F(ab′) 2 coated on the assay plate used to capture receptor-hexaHis tail fusion or anti-GST antibody coated on the assay plate used to capture a receptor-GST fusion.
  • binding to cells expressing at least the binding domain of the receptor may be evaluated.
  • the cells may be naturally occurring hematopoietic cells that express the FcR of interest or may be transformed with nucleic acid encoding the FcR or a binding domain thereof such that the binding domain is expressed at the surface of the cell to be tested.
  • the immune complex described hereinabove is added to the receptor-coated plates and incubated for a sufficient period of time such that the analyte binds to the receptor. Plates may then be washed to remove unbound complexes, and binding of the analyte may be detected according to known methods. For example, binding may be detected using a reagent (e.g. an antibody or fragment thereof) which binds specifically to the analyte, and which is optionally conjugated with a detectable label (detectable labels and methods for conjugating them to polypeptides are described below in the section entitled “Non-Therapeutic Uses for the Polypeptide Variant”).
  • a reagent e.g. an antibody or fragment thereof
  • the reagents can be provided in an assay kit, i.e., a packaged combination of reagents, for combination with the analyte in assaying the ability of the analyte to bind to a receptor of interest.
  • the components of the kit will generally be provided in predetermined ratios.
  • the kit may provide the first target molecule and/or the second target molecule, optionally complexed together.
  • the kit may further include assay plates coated with the receptor or a binding domain thereof (e.g. the extracellular domain of the ⁇ subunit of an FcR).
  • kits such as an antibody that binds specifically to the analyte to be assayed, labeled directly or indirectly with an enzymatic label
  • the detectable label is an enzyme
  • the kit will include substrates and cofactors required by the enzyme (e.g. a substrate precursor which provides the detectable chromophore or fluorophore).
  • other additives may be included such as stabilizers, buffers (e.g. assay and/or wash lysis buffer) and the like.
  • the relative amounts of the various reagents may be varied widely to provide for concentrations in solution of the reagents that substantially optimize the sensitivity of the assay.
  • the reagents may be provided as dry powders, usually lyophilized, including excipients that on dissolution will provide a reagent solution having the appropriate concentration.
  • the kit also suitably includes instructions for carrying out the assay.
  • the Fc region-containing polypeptide which is modified according to the teachings herein is an antibody. Techniques for producing antibodies follow:
  • the polypeptide is an antibody
  • it is directed against an antigen of interest.
  • the antigen is a biologically important polypeptide and administration of the antibody to a mammal suffering from a disease or disorder can result in a therapeutic benefit in that mammal.
  • antibodies directed against nonpolypeptide antigens are also contemplated.
  • the antigen is a polypeptide, it may be a transmembrane molecule (e.g. receptor) or ligand such as a growth factor.
  • exemplary antigens include molecules such as renin; a growth hormone, including human growth hormone and bovine growth hormone; growth hormone releasing factor; parathyroid hormone; thyroid stimulating hormone; lipoproteins; alpha-1-antitrypsin; insulin A-chain; insulin B-chain; proinsulin; follicle stimulating hormone; calcitonin; luteinizing hormone; glucagon; clofting factors such as factor VIIIC, factor IX, tissue factor, and von Willebrands factor; anti-clofting factors such as Protein C; atrial natriuretic factor; lung surfactant; a plasminogen activator, such as urokinase or human urine or tissue-type plasminogen activator (t-PA); bombesin; thrombin; hemopoietic growth factor; tumor necrosis factor-alpha
  • Preferred molecular targets for antibodies encompassed by the present invention include CD proteins such as CD3, CD4, CD8, CD19, CD20 and CD34; members of the ErbB receptor family such as the EGF receptor, HER2, HER3 or HER4 receptor; cell adhesion molecules such as LFA-1, Mac1, p150.95, VLA-4, ICAM-1, VCAM and ⁇ v/ ⁇ 3 integrin including either ⁇ or ⁇ subunits thereof (e.g. anti-CD11 a , anti-CD18 or anti-CD11 b antibodies); growth factors such as VEGF; IgE; blood group antigens; flk2/flt3 receptor; obesity (OB) receptor; mpl receptor; CTLA-4; protein C etc.
  • CD proteins such as CD3, CD4, CD8, CD19, CD20 and CD34
  • members of the ErbB receptor family such as the EGF receptor, HER2, HER3 or HER4 receptor
  • cell adhesion molecules such as LFA-1, Mac1, p150.95
  • Soluble antigens or fragments thereof, optionally conjugated to other molecules, can be used as immunogens for generating antibodies.
  • immunogens for transmembrane molecules, such as receptors, fragments of these (e.g. the extracellular domain of a receptor) can be used as the immunogen.
  • transmembrane molecules such as receptors
  • fragments of these e.g. the extracellular domain of a receptor
  • cells expressing the transmembrane molecule can be used as the immunogen.
  • Such cells can be derived from a natural source (e.g. cancer cell lines) or may be cells which have been transformed by recombinant techniques to express the transmembrane molecule.
  • Other antigens and forms thereof useful for preparing antibodies will be apparent to those in the art.
  • Polyclonal antibodies are preferably raised in animals by multiple subcutaneous (sc) or intraperitoneal (ip) injections of the relevant antigen and an adjuvant. It may be useful to conjugate the relevant antigen to a protein that is immunogenic in the species to be immunized, e.g., keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, or soybean trypsin inhibitor using a bifunctional or derivatizing agent, for example, maleimidobenzoyl sulfosuccinimide ester (conjugation through cysteine residues), N-hydroxysuccinimide (through lysine residues), glutaraldehyde, succinic anhydride, SOCl 2 , or R 1 N ⁇ C ⁇ NR, where R and R 1 are different alkyl groups.
  • a protein that is immunogenic in the species to be immunized e.g., keyhole limpet hemocyanin, serum albumin, bovine thy
  • Animals are immunized against the antigen, immunogenic conjugates, or derivatives by combining, e.g., 100 ⁇ g or 5 ⁇ g of the protein or conjugate (for rabbits or mice, respectively) with 3 volumes of Freund's complete adjuvant and injecting the solution intradermally at multiple sites.
  • the animals are boosted with 1/5 to 1/10 the original amount of peptide or conjugate in Freund's complete adjuvant by subcutaneous injection at multiple sites.
  • Seven to 14 days later the animals are bled and the serum is assayed for antibody titer. Animals are boosted until the titer plateaus.
  • the animal is boosted with the conjugate of the same antigen, but conjugated to a different protein and/or through a different cross-linking reagent.
  • Conjugates also can be made in recombinant cell culture as protein fusions.
  • aggregating agents such as alum are suitably used to enhance the immune response.
  • Monoclonal antibodies may be made using the hybridoma method first described by Kohler et al, Nature, 256:495 (1975), or may be made by recombinant DNA methods (U.S. Pat. No. 4,816,567).
  • a mouse or other appropriate host animal such as a hamster or macaque monkey
  • lymphocytes that produce or are capable of producing antibodies that will specifically bind to the protein used for immunization.
  • lymphocytes may be immunized in vitro. Lymphocytes then are fused with myeloma cells using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell (Goding, Monoclonal Antibodies: Principles and Practice , pp.59-103 (Academic Press, 1986)).
  • the hybridoma cells thus prepared are seeded and grown in a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, parental myeloma cells.
  • a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, parental myeloma cells.
  • the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine (HAT medium), which substances prevent the growth of HGPRT-deficient cells.
  • Preferred myeloma cells are those that fuse efficiently, support stable high-level production of antibody by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium.
  • preferred myeloma cell lines are murine myeloma lines, such as those derived from MOPC-21 and MPC-11 mouse tumors available from the Salk Institute Cell Distribution Center, San Diego, Calif. USA, and SP-2 or X63-Ag8-653 cells available from the American Type Culture Collection, Rockville, Md. USA.
  • Human myeloma and mouse-human heteromyeloma cell lines also have been described for the production of human monoclonal antibodies (Kozbor, J. Immunol ., 133:3001 (1984); Brodeur et al, Monoclonal Antibody Production Techniques and Applications , pp. 51-63 (Marcel Dekker, Inc., New York, 1987)).
  • Culture medium in which hybridoma cells are growing is assayed for production of monoclonal antibodies directed against the antigen.
  • the binding specificity of monoclonal antibodies produced by hybridoma cells is determined by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA).
  • RIA radioimmunoassay
  • ELISA enzyme-linked immunoabsorbent assay
  • the clones may be subcloned by limiting dilution procedures and grown by standard methods (Goding, Monoclonal Antibodies: Principles and Practice , pp.59-103 (Academic Press, 1986)). Suitable culture media for this purpose include, for example, D-MEM or RPMI-1640 medium.
  • the hybridoma cells may be grown in vivo as ascites tumors in an animal.
  • the monoclonal antibodies secreted by the subclones are suitably separated from the culture medium, ascites fluid, or serum by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxylapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.
  • DNA encoding the monoclonal antibodies is readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of the monoclonal antibodies).
  • the hybridoma cells serve as a preferred source of such DNA.
  • the DNA may be placed into expression vectors, which are then transfected into host cells such as E. coi cells, simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin protein, to obtain the synthesis of monoclonal antibodies in the recombinant host cells. Recombinant production of antibodies will be described in more detail below.
  • antibodies or antibody fragments can be isolated from antibody phage libraries generated using the techniques described in McCafferty et al, Nature , 348:552-554 1990). Clackson et al., Nature , 352:624-628 (1991) and Marks et al, J. Mol. Biol ., 222:581-597 (1991) describe the isolation of murine and human antibodies, respectively, using phage libraries.
  • the DNA also may be modified, for example, by substituting the coding sequence for human heavy- and light-chain constant domains in place of the homologous murine sequences (U.S. Pat. No. 4,816,567; Morrison, et al., Proc. Natl Acad. Sci. USA , 81:6851 (1984)), or by covalently joining to the immunoglobulin coding sequence all or part of the coding sequence for a non-immunoglobulin polypeptide.
  • non-immunoglobulin polypeptides are substituted for the constant domains of an antibody, or they are substituted for the variable domains of one antigen-combining site of an antibody to create a chimeric bivalent antibody comprising one antigen-combining site having specificity for an antigen and another antigen-combining site having specificity for a different antigen.
  • a humanized antibody has one or more amino acid residues introduced into it from a source which is non-human. These non-human amino acid residues are often referred to as “import” residues, which are typically taken from an “import” variable domain. Humanization can be essentially performed following the method of Winter and co-workers (Jones et al., Nature, 321:522-525 (1986); Riechmann et al, Nature , 332:323-327 (1988); Verhoeyen et al, Science , 239:1534-1536 (1988)), by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody. Accordingly, such “humanized” antibodies are chimeric antibodies (U.S.
  • humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR residues are substituted by residues from analogous sites in rodent antibodies.
  • variable domains both light and heavy
  • the choice of human variable domains, both light and heavy, to be used in making the humanized antibodies is very important to reduce antigenicity.
  • the sequence of the variable domain of a rodent antibody is screened against the entire library of known human variable-domain sequences.
  • the human sequence which is closest to that of the rodent is then accepted as the human framework (FR) for the humanized antibody (Sims et al, J. Immunol ., 151:2296 (1993); Chothia et al., J. Mol Biol , 196:901 (1987)).
  • Another method uses a particular framework derived from the consensus sequence of all human antibodies of a particular subgroup of light or heavy chains.
  • the same framework may be used for several different humanized antibodies (Carter et al., Proc. Natl. Acad. Sci. USA , 89:4285 (1992); Presta et al., J. Immnol ., 151:2623 (1993)).
  • humanized antibodies are prepared by a process of analysis of the parental sequences and various conceptual humanized products using three-dimensional models of the parental and humanized sequences.
  • Three-dimensional immunoglobulin models are commonly available and are familiar to those skilled in the art.
  • Computer programs are available which illustrate and display probable three-dimensional conformational structures of selected candidate immunoglobulin sequences. Inspection of these displays permits analysis of the likely role of the residues in the functioning of the candidate immunoglobulin sequence, i.e., the analysis of residues that influence the ability of the candidate immunoglobulin to bind its antigen.
  • FR residues can be selected and combined from the recipient and import sequences so that the desired antibody characteristic, such as increased affinity for the target antigen(s), is achieved.
  • the CDR residues are directly and most substantially involved in influencing antigen binding.
  • transgenic animals e.g., mice
  • transgenic animals e.g., mice
  • J H antibody heavy-chain joining region
  • Human antibodies can also be derived from phage-display libraries (Hoogenboom et al., J. MoL Biol ., 227:381 (1991); Marks et al., J. Mol. Biol , 222:581-597 (1991); Vaughan et al. Nature Biotech 14:309 (1996)).
  • Multispecific antibodies have binding specificities for at least two different antigens. While such molecules normally will only bind two antigens (i.e. bispecific antibodies, BsAbs), antibodies with additional specificities such as trispecific antibodies are encompassed by this expression when used herein.
  • BsAbs include those with one arm directed against a tumor cell antigen and the other arm directed against a cytotoxic trigger molecule such as anti-Fc ⁇ Rl/anti-CD15, anti-p185 HER2 /Fc ⁇ RIII (CD16), anti-CD3/anti-malignant B-cell (1D10), anti-CD3/anti-p185 HER2 , anti-CD3/anti-p97, anti-CD3/anti-renal cell carcinoma, anti-CD3/anti-OVCAR-3, anti-CD3/L-D1 (anti-colon carcinoma), anti-CD3/anti-melanocyte stimulating hormone analog, anti-EGF receptor/anti-CD3, anti-CD3/anti-CAMA1, anti-CD3/anti-CD19, anti-CD3/MoV18, anti-neural cell ahesion molecule (NCAM)/anti-CD3, anti-folate binding protein (FBP)/anti-CD3, anti-pan carcinoma associated antigen (AMOC-31)/anti-CD3; BsAbs with one arm which bind
  • BsAbs for use in therapy of infectious diseases such as anti-CD3/anti-herpes simplex virus (HSV), anti-T-cell receptor:CD3 complex/anti-influenza, anti-Fc ⁇ R/anti-HIV; BsAbs for tumor detection in vitro or in vivo such as anti-CEA/anti-EOTUBE, anti-CEA/anti-DPTA, anti-p185 HER2 /anti-hapten; BsAbs as vaccine adjuvants; and BsAbs as diagnostic tools such as anti-rabbit IgG/anti-ferritin, anti-horse radish peroxidase (HRP)/anti-hormone, anti-somatostatin/anti-substance P, anti-HRP/anti-FITC, anti-CEA/anti- ⁇ -galactosidase.
  • HRP anti-horse radish peroxidase
  • HRP anti-somatostatin/anti-substance P
  • trispecific antibodies examples include anti-CD3/anti-CD4/anti-CD37, anti-CD3/anti-CD5/anti-CD37 and anti-CD3/anti-CD8/anti-CD37.
  • Bispecific antibodies can be prepared as full length antibodies or antibody fragments (e.g. F(ab′) 2 bispecific antibodies).
  • antibody variable domains with the desired binding specificities are fused to immunoglobulin constant domain sequences.
  • the fusion preferably is with an immunoglobulin heavy chain constant domain, comprising at least part of the hinge, CH2, and CH3 regions. It is preferred to have the first heavy-chain constant region (CH1) containing the site necessary for light chain binding, present in at least one of the fusions.
  • DNAs encoding the immunoglobulin heavy chain fusions and, if desired, the immunoglobulin light chain are inserted into separate expression vectors, and are co-transfected into a suitable host organism.
  • the bispecific antibodies are composed of a hybrid immunoglobulin heavy chain with a first binding specificity in one arm, and a hybrid immunoglobulin heavy chain-light chain pair (providing a second binding specificity) in the other arm. It was found that this asymmetric structure facilitates the separation of the desired bispecific compound from unwanted immunoglobulin chain combinations, as the presence of an immunoglobulin light chain in only one half of the bispecific molecule provides for a facile way of separation. This approach is disclosed in WO 94/04690. For further details of generating bispecific antibodies see, for example, Suresh et al., Methods in Enzymology , 121:210 (1986).
  • the interface between a pair of antibody molecules can be engineered to maximize the percentage of heterodimers which are recovered from recombinant cell culture.
  • the preferred interface comprises at least a part of the C H 3 domain of an antibody constant domain.
  • one or more small amino acid side chains from the interface of the first antibody molecule are replaced with larger side chains (e.g. tyrosine or tryptophan).
  • Compensatory “cavities” of identical or similar size to the large side chain(s) are created on the interface of the second antibody molecule by replacing large amino acid side chains with smaller ones (e.g. alanine or threonine). This provides a mechanism for increasing the yield of the heterodimer over other unwanted end-products such as homodimers.
  • Bispecific antibodies include cross-linked or “heteroconjugate” antibodies.
  • one of the antibodies in the heteroconjugate can be coupled to avidin, the other to biotin.
  • Such antibodies have, for example, been proposed to target immune system cells to unwanted cells (U.S. Pat. No. 4,676,980), and for treatment of HIV infection (WO 91/00360, WO 921200373, and EP 03089).
  • Heteroconjugate antibodies may be made using any convenient cross-linking methods. Suitable cross-linking agents are well known in the art, and are disclosed in U.S. Pat. No. 4,676,980, along with a number of cross-linking techniques.
  • Antibodies with more than two valencies are contemplated.
  • trispecific antibodies can be prepared. Tutt et al J. Immunol . 147:60 (1991).
  • polypeptide of interest herein is preferably an antibody
  • Fc region-containing polypeptides which can be modified according to the methods described herein are contemplated.
  • An example of such a molecule is an immunoadhesin.
  • the simplest and most straightforward immunoadhesin design combines the binding domain(s) of the adhesin (e.g. the extracellular domain (ECD) of a receptor) with the Fc region of an immunoglobulin heavy chain.
  • ECD extracellular domain
  • nucleic acid encoding the binding domain of the adhesin will be fused C-terminally to nucleic acid encoding the N-terminus of an immunoglobulin constant domain sequence, however N-terminal fusions are also possible.
  • the encoded chimeric polypeptide will retain at least functionally active hinge, C H 2 and C H 3 domains of the constant region of an immunoglobulin heavy chain. Fusions are also made to the C-terminus of the Fc portion of a constant domain, or immediately N-terminal to the C H 1 of the heavy chain or the corresponding region of the light chain.
  • the precise site at which the fusion is made is not critical; particular sites are well known and may be selected in order to optimize the biological activity, secretion, or binding characteristics of the immunoadhesin.
  • the adhesin sequence is fused to the N-terminus of the Fc region of immunoglobulin G 1 (IgG 1 ). It is possible to fuse the entire heavy chain constant region to the adhesin sequence. However, more preferably, a sequence beginning in the hinge region just upstream of the papain cleavage site which defines IgG Fc chemically (i.e. residue 216, taking the first residue of heavy chain constant region to be 114), or analogous sites of other immunoglobulins is used in the fusion.
  • the adhesin amino acid sequence is fused to (a) the hinge region and C H 2 and C H 3 or (b) the C H 1, hinge, C H 2 and C H 3 domains, of an IgG heavy chain.
  • the immunoadhesins are assembled as multimers, and particularly as heterodimers or heterotetramers.
  • these assembled immunoglobulins will have known unit structures.
  • a basic four chain structural unit is the form in which IgG, lgD, and IgE exist.
  • a four chain unit is repeated in the higher molecular weight immunoglobulins; IgM generally exists as a pentamer of four basic units held together by disulfide bonds.
  • IgA globulin, and occasionally IgG globulin may also exist in multimeric form in serum. In the case of multimer, each of the four units may be the same or different.
  • each A represents identical or different adhesin amino acid sequences
  • V L is an immunoglobulin light chain variable domain
  • V H is an immunoglobulin heavy chain variable domain
  • C L is an immunoglobulin light chain constant domain
  • C H is an immunoglobulin heavy chain constant domain
  • n is an integer greater than 1;
  • Y designates the residue of a covalent cross-linking agent.
  • the adhesin sequences can be inserted between immunoglobulin heavy chain and light chain sequences, such that an immunoglobulin comprising a chimeric heavy chain is obtained.
  • the adhesin sequences are fused to the 3′ end of an immunoglobulin heavy chain in each arm of an immunoglobulin, either between the hinge and the C H 2 domain, or between the C H 2 and C H 3 domains. Similar constructs have been reported by Hoogenboom, et al., Mol. Immunol . 28:1027-1037 (1991).
  • an immunoglobulin light chain might be present either covalently associated to an adhesin-immunoglobulin heavy chain fusion polypeptide, or directly fused to the adhesin.
  • DNA encoding an immunoglobulin light chain is typically coexpressed with the DNA encoding the adhesin-immunoglobulin heavy chain fusion protein.
  • the hybrid heavy chain and the light chain will be covalently associated to provide an immunoglobulin-like structure comprising two disulfide-linked immunoglobulin heavy chain-light chain pairs.
  • lmmunoadhesins are most conveniently constructed by fusing the cDNA sequence encoding the adhesin portion in-frame to an immunoglobulin cDNA sequence.
  • fusion to genomic immunoglobulin fragments can also be used (see, e.g. Aruffo et al, Cell 61:1303-1313 (1990); and Stamenkovic et al, Cell 66:1133-1144 (1991)).
  • the latter type of fusion requires the presence of Ig regulatory sequences for expression.
  • cDNAs encoding IgG heavy-chain constant regions can be isolated based on published sequences from cDNA libraries derived from spleen or peripheral blood lymphocytes, by hybridization or by polymerase chain reaction (PCR) techniques.
  • the cDNAs encoding the “adhesin” and the immunoglobulin parts of the immunoadhesin are inserted in tandem into a plasmid vector that directs efficient expression in the chosen host cells.
  • the invention also provides isolated nucleic acid encoding a polypeptide variant as disclosed herein, vectors and host cells comprising the nucleic acid, and recombinant techniques for the production of the polypeptide variant.
  • the nucleic acid encoding it is isolated and inserted into a replicable vector for further cloning (amplification of the DNA) or for expression.
  • DNA encoding the polypeptide variant is readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the polypeptide variant).
  • Many vectors are available.
  • the vector components generally include, but are not limited to, one or more of the following: a signal sequence, an origin of replication, one or more marker genes, an enhancer element, a promoter, and a transcription termination sequence.
  • the polypeptide variant of this invention may be produced recombinantly not only directly, but also as a fusion polypeptide with a heterologous polypeptide, which is preferably a signal sequence or other polypeptide having a specific cleavage site at the N-terminus of the mature protein or polypeptide.
  • a heterologous polypeptide which is preferably a signal sequence or other polypeptide having a specific cleavage site at the N-terminus of the mature protein or polypeptide.
  • the heterologous signal sequence selected preferably is one that is recognized and processed (ie., cleaved by a signal peptidase) by the host cell.
  • the signal sequence is substituted by a prokaryotic signal sequence selected, for example, from the group of the alkaline phosphatase, penicillinase, Ipp, or heat-stable enterotoxin II leaders.
  • a prokaryotic signal sequence selected, for example, from the group of the alkaline phosphatase, penicillinase, Ipp, or heat-stable enterotoxin II leaders.
  • yeast secretion the native signal sequence may be substituted by, e.g., the yeast invertase leader, ⁇ factor leader (including Saccharomyces and Kluyveromyces ⁇ -factor leaders), or acid phosphatase leader, the C. albicans glucoamylase leader, or the signal described in WO 90113646.
  • mammalian signal sequences as well as viral secretory leaders for example, the herpes simplex gD signal, are available.
  • the DNA for such precursor region is ligated in reading frame to DNA encoding the polypeptide variant.
  • Origin of Replication Component Both Expression and Cloning Vectors Contain a nucleic acid sequence that enables the vector to replicate in one or more selected host cells.
  • this sequence is one that enables the vector to replicate independently of the host chromosomal DNA, and includes origins of replication or autonomously replicating sequences.
  • origins of replication or autonomously replicating sequences are well known for a variety of bacteria, yeast, and viruses.
  • the origin of replication from the plasmid pBR322 is suitable for most Gram-negative bacteria, the 2 ⁇ plasmid origin is suitable for yeast, and various viral origins (SV40, polyoma, adenovirus, VSV or BPV) are useful for cloning vectors in mammalian cells.
  • the origin of replication component is not needed for mammalian expression vectors (the SV40 origin may typically be used only because it contains the early promoter).
  • Expression and cloning vectors may contain a selection gene, also termed a selectable marker.
  • Typical selection genes encode proteins that (a) confer resistance to antibiotics or other toxins, e.g., ampicillin, neomycin, methotrexate, or tetracycline, (b) complement auxotrophic deficiencies, or (c) supply critical nutrients not available from complex media, e.g., the gene encoding D-alanine racemase for Bacilli.
  • One example of a selection scheme utilizes a drug to arrest growth of a host cell. Those cells that are successfully transformed with a heterologous gene produce a protein conferring drug resistance and thus survive the selection regimen. Examples of such dominant selection use the drugs neomycin, mycophenolic acid and hygromycin.
  • Suitable selectable markers for mammalian cells are those that enable the identification of cells competent to take up the polypeptide variant nucleic acid, such as DHFR, thymidine kinase, metallothionein-I and -II, preferably primate metallothionein genes, adenosine deaminase, ornithine decarboxylase, etc.
  • cells transformed with the DHFR selection gene are first identified by culturing all of the transformants in a culture medium that contains methotrexate (Mtx), a competitive antagonist of DHFR.
  • Mtx methotrexate
  • An appropriate host cell when wild-type DHFR is employed is the Chinese hamster ovary (CHO) cell line deficient in DHFR activity.
  • host cells transformed or co-transformed with DNA sequences encoding polypeptide variant, wild-type DHFR protein, and another selectable marker such as aminoglycoside 3′-phosphotransferase (APH) can be selected by cell growth in medium containing a selection agent for the selectable marker such as an aminoglycosidic antibiotic, e.g., kanamycin, neomycin, or G418. See U.S. Pat. No. 4,965,199.
  • APH aminoglycoside 3′-phosphotransferase
  • a suitable selection gene for use in yeast is the trpl gene present in the yeast plasmid YRp7 (Stinchcomb et al., Nature , 282:39 (1979)).
  • the trpl gene provides a selection marker for a mutant strain of yeast lacking the ability to grow in tryptophan, for example, ATCC No. 44076 or PEP4-1. Jones, Genetics, 85:12 (1977).
  • the presence of the trp1 lesion in the yeast host cell genome then provides an effective environment for detecting transformation by growth in the absence of tryptophan.
  • Leu2-deficient yeast strains (ATCC 20,622 or 38,626) are complemented by known plasmids bearing the Leu2 gene.
  • vectors derived from the 1.6 ⁇ m circular plasmid pKDl can be used for transformation of Kluyveromyces yeasts.
  • an expression system for large-scale production of recombinant calf chymosin was reported for K. lactis. Van den Berg, Bio/Technology , 8:135 (1990).
  • Stable multi-copy expression vectors for secretion of mature recombinant human serum albumin by industrial strains of Kluyveromyces have also been disclosed. Fleer et al., Bio/Technology , 9:968-975 (1991).
  • Expression and cloning vectors usually contain a promoter that is recognized by the host organism and is operably linked to the polypeptide variant nucleic acid.
  • Promoters suitable for use with prokaryotic hosts include the phoA promoter, ⁇ -lactamase and lactose promoter systems, alkaline phosphatase, a tryptophan (trp) promoter system, and hybrid promoters such as the tac promoter.
  • trp tryptophan
  • Other known bacterial promoters are suitable. Promoters for use in bacterial systems also will contain a Shine-Dalgarno (S.D.) sequence operably linked to the DNA encoding the polypeptide variant.
  • S.D. Shine-Dalgarno
  • Promoter sequences are known for eukaryotes. Virtually all eukaryotic genes have an AT-rich region located approximately 25 to 30 bases upstream from the site where transcription is initiated. Another sequence found 70 to 80 bases upstream from the start of transcription of many genes is a CNCAAT region where N may be any nucleotide. At the 3′ end of most eukaryotic genes is an AATAAA sequence that may be the signal for addition of the poly A tail to the 3′ end of the coding sequence. All of these sequences are suitably inserted into eukaryotic expression vectors.
  • suitable promoting sequences for use with yeast hosts include the promoters for 3-phosphoglycerate kinase or other glycolytic enzymes, such as enolase, glyceraldehyde-3-phosphate dehydrogenase, hexokinase, pyruvate decarboxylase, phosphofructokinase, glucose-6-phosphate isomerase, 3-phosphoglycerate mutase, pyruvate kinase, triosephosphate isomerase, phosphoglucose isomerase, and glucokinase.
  • 3-phosphoglycerate kinase or other glycolytic enzymes such as enolase, glyceraldehyde-3-phosphate dehydrogenase, hexokinase, pyruvate decarboxylase, phosphofructokinase, glucose-6-phosphate isomerase, 3-phosphoglycerate mutase, pyruvate
  • yeast promoters which are inducible promoters having the additional advantage of transcription controlled by growth conditions, are the promoter regions for alcohol dehydrogenase 2, isocytochrome C, acid phosphatase, degradative enzymes associated with nitrogen metabolism, metallothionein, glyceraldehyde-3-phosphate dehydrogenase, and enzymes responsible for maltose and galactose utilization.
  • Suitable vectors and promoters for use in yeast expression are further described in EP 73,657.
  • Yeast enhancers also are advantageously used with yeast promoters.
  • Polypeptide variant transcription from vectors in mammalian host cells is controlled, for example, by promoters obtained from the genomes of viruses such as polyoma virus, fowlpox virus, adenovirus (such as Adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-B virus and most preferably Simian Virus 40 (SV40), from heterologous mammalian promoters, e.g., the actin promoter or an immunoglobulin promoter, from heat-shock promoters, provided such promoters are compatible with the host cell systems.
  • viruses such as polyoma virus, fowlpox virus, adenovirus (such as Adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-B virus and most preferably Simian Virus 40 (
  • the early and late promoters of the SV40 virus are conveniently obtained as an SV40 restriction fragment that also contains the SV40 viral origin of replication.
  • the immediate early promoter of the human cytomegalovirus is conveniently obtained as a HindIII E restriction fragment.
  • a system for expressing DNA in mammalian hosts using the bovine papilloma virus as a vector is disclosed in U.S. Pat. No. 4,419,446. A modification of this system is described in U.S. Pat. No. 4,601,978.
  • Enhancer sequences are now known from mammalian genes (globin, elastase, albumin, ⁇ -fetoprotein, and insulin). Typically, however, one will use an enhancer from a eukaryotic cell virus. Examples include the SV40 enhancer on the late side of the replication origin (bp 100-270), the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, and adenovirus enhancers.
  • the enhancer may be spliced into the vector at a position 5′ or 3′ to the polypeptide variant-encoding sequence, but is preferably located at a site 5′ from the promoter.
  • Expression vectors used in eukaryotic host cells will also contain sequences necessary for the termination of transcription and for stabilizing the mRNA. Such sequences are commonly available from the 5′ and, occasionally 3′ , untranslated regions of eukaryotic or viral DNAs or cDNAs. These regions contain nucleotide segments transcribed as polyadenylated fragments in the untranslated portion of the mRNA encoding the polypeptide variant.
  • One useful transcription termination component is the bovine growth hormone polyadenylation region. See WO94/11026 and the expression vector disclosed therein.
  • Suitable host cells for cloning or expressing the DNA in the vectors herein are the prokaryote, yeast, or higher eukaryote cells described above.
  • Suitable prokaryotes for this purpose include eubacteria, such as Gram-negative or Gram-positive organisms, for example, Enterobacteriaceae such as Escherichia, e.g., E coli , Enterobacter, Erwinia, Klebsiella, Proteus, Salmonella, e.g., Salmonella typhimurium , Serratia, e.g., Serratia marcescans , and Shigella, as well as Bacilli such as B. subtilis and B.
  • Enterobacteriaceae such as Escherichia, e.g., E coli , Enterobacter, Erwinia, Klebsiella, Proteus
  • Salmonella e.g., Salmonella typhimurium
  • Serratia e.g
  • E. coil cloning host is E. coli 294 (ATCC 31,446), although other strains such as E. coli B, E. coil X1776 (ATCC 31,537), and E. coli W3110 (ATCC 27,325) are suitable. These examples are illustrative rather than limiting.
  • eukaryotic microbes such as filamentous fungi or yeast are suitable cloning or expression hosts for polypeptide variant-encoding vectors.
  • Saccharomyces cerevisiae or common baker's yeast, is the most commonly used among lower eukaryotic host microorganisms.
  • a number of other genera, species, and strains are commonly available and useful herein, such as Schizosaccharomyces pombe; Kluyveromyces hosts such as, e.g., K lactis, K. fragilis (ATCC 12,424), K. bulgaricus (ATCC 16,045), K. wickeramii (ATCC 24,178), K.
  • waltii ATCC 56,500
  • K. drosophilarum ATCC 36,906
  • K. thermotolerans K. marxianus
  • yarrowia EP 402,226
  • Pichia pastoris EP 183,070
  • Candida Trichoderma reesia
  • Neurospora crassa Schwanniomyces such as Schwanniomyces occidentalis
  • filamentous fungi such as, e.g., Neurospora, Penicillium, Tolypocladium, and Aspergillus hosts such as A. nidulans and A. niger .
  • Suitable host cells for the expression of glycosylated polypeptide variant are derived from multicellular organisms.
  • invertebrate cells include plant and insect cells.
  • Numerous baculoviral strains and variants and corresponding permissive insect host cells from hosts such as Spodoptera frugiperda (caterpillar), Aedes aegypti (mosquito), Aedes albopictus (mosquito), Drosophila melanogaster (fruiffly), and Bombyx mori have been identified.
  • a variety of viral strains for transfection are publicly available, e.g., the L-1 variant of Autographa californica NPV and the Bm-5 strain of Bombyx mori NPV, and such viruses may be used as the virus herein according to the present invention, particularly for transfection of Spodoptera frugiperda cells.
  • Plant cell cultures of cotton, corn, potato, soybean, petunia, tomato, and tobacco can also be utilized as hosts.
  • vertebrate cells have been greatest in vertebrate cells, and propagation of vertebrate cells in culture (tissue culture) has become a routine procedure.
  • useful mammalian host cell lines are monkey kidney CV1 line transformed by SV40 (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture, Graham et al., J. Gen Virol . 36:59 (1977)); baby hamster kidney cells (BHK, ATCC CCL 10); Chinese hamster ovary cells/-DHFR (CHO, Urlaub et al, Proc. Natl. Acad. Sci. USA 77:4216 (1980)); mouse sertoli cells (TM4, Mather, Biol. Reprod .
  • COS-7 monkey kidney CV1 line transformed by SV40
  • human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture, Graham et al., J. Gen Virol . 36:59 (1977)
  • monkey kidney cells (CV1 ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL-1587); human cervical carcinoma cells (HELA, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); buffalo rat liver cells (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells (Hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL51); TRI cells (Mather et al., Annals N.Y. Acad. Sci . 383:44-68 (1982)); MRC 5 cells; FS4 cells; and a human hepatoma line (Hep G2).
  • Host cells are transformed with the above-described expression or cloning vectors for polypeptide variant production and cultured in conventional nutrient media modified as appropriate for inducing promoters, selecting transformants, or amplifying the genes encoding the desired sequences.
  • the host cells used to produce the polypeptide variant of this invention may be cultured in a variety of media.
  • Commercially available media such as Ham's F10 (Sigma), Minimal Essential Medium ((MEM), (Sigma), RPMI-1640 (Sigma), and Dulbecco's Modified Eagle's Medium ((DMEM), Sigma) are suitable for culturing the host cells.
  • any of these media may be supplemented as necessary with hormones and/or other growth factors (such as insulin, transferrin, or epidermal growth factor), salts (such as sodium chloride, calcium, magnesium, and phosphate), buffers (such as HEPES), nucleotides (such as adenosine and thymidine), antibiotics (such as GENTAMYCINTM drug), trace elements (defined as inorganic compounds usually present at final concentrations in the micromolar range), and glucose or an equivalent energy source. Any other necessary supplements may also be included at appropriate concentrations that would be known to those skilled in the art.
  • the culture conditions such as temperature, pH, and the like, are those previously used with the host cell selected for expression, and will be apparent to the ordinarily skilled artisan.
  • the polypeptide variant can be produced intracellularly, in the periplasmic space, or directly secreted into the medium. If the polypeptide variant is produced intracellularly, as a first step, the particulate debris, either host cells or lysed fragments, is removed, for example, by centrifugation or ultrafiltration. Carter et al., Bio/lTechnology 10:163-167 (1992) describe a procedure for isolating antibodies which are secreted to the periplasmic space of E coil .
  • cell paste is thawed in the presence of sodium acetate (pH 3.5), EDTA, and phenylmethylsulfonylfluoride (PMSF) over about 30 min.
  • PMSF phenylmethylsulfonylfluoride
  • Cell debris can be removed by centrifugation.
  • supernatants from such expression systems are generally first concentrated using a commercially available protein concentration filter, for example, an Amicon or Millipore Pellicon ultrafiltration unit.
  • a protease inhibitor such as PMSF may be included in any of the foregoing steps to inhibit proteolysis and antibiotics may be included to prevent the growth of adventitious contaminants.
  • the polypeptide variant composition prepared from the cells can be purified using, for example, hydroxylapatite chromatography, gel electrophoresis, dialysis, and affinity chromatography, with affinity chromatography being the preferred purification technique.
  • affinity chromatography is the preferred purification technique.
  • the suitability of protein A as an affinity ligand depends on the species and isotype of any immunoglobulin Fc region that is present in the polypeptide variant. Protein A can be used to purify polypeptide variants that are based on human ⁇ 1, ⁇ 2, or ⁇ 4 heavy chains (Lindmark et al, J. Immunol. Meth . 62:1-13 (1983)).
  • Protein G is recommended for all mouse isotypes and for human ⁇ 3 (Guss et al, EMBO J . 5:15671575 (1986)).
  • the matrix to which the affinity ligand is attached is most often agarose, but other matrices are available. Mechanically stable matrices such as controlled pore glass or poly(styrenedivinyl)benzene allow for faster flow rates and shorter processing times than can be achieved with agarose.
  • the polypeptide variant comprises a C H 3 domain
  • the Bakerbond ABXTM resin J. T. Baker, Phillipsburg, N.J.
  • the mixture comprising the polypeptide variant of interest and contaminants may be subjected to low pH hydrophobic interaction chromatography using an elution buffer at a pH between about 2.5-4.5, preferably performed at low salt concentrations (e.g.,from about 0-0.25M salt).
  • Therapeutic formulations of the polypeptide variant are prepared for storage by mixing the polypeptide variant having the desired degree of purity with optional physiologically acceptable carriers, excipients or stabilizers ( Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)), in the form of lyophilized formulations or aqueous solutions.
  • Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyidimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptide; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arg
  • the formulation herein may also contain more than one active compound as necessary for the particular indication being treated, preferably those with complementary activities that do not adversely affect each other.
  • Such molecules are suitably present in combination in amounts that are effective for the purpose intended.
  • the active ingredients may also be entrapped in microcapsule prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsule and poly-(methylmethacylate) microcapsule, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules) or in macroemulsions.
  • colloidal drug delivery systems for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules
  • formulations to be used for in vivo administration must be sterile. This is readily accomplished by filtration through sterile filtration membranes.
  • Sustained-release preparations may be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the polypeptide variant, which matrices are in the form of shaped articles, e.g., films, or microcapsule. Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate), or poly(vinylalcohol)), polylactides (U.S. Pat. No.
  • copolymers of L-glutamic acid and ⁇ ethyl-L-glutamate copolymers of L-glutamic acid and ⁇ ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the LUPRON DEPOTTM (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-( ⁇ )-3-hydroxybutyric acid. While polymers such as ethylene-vinyl acetate and lactic acid-glycolic acid enable release of molecules for over 100 days, certain hydrogels release proteins for shorter time periods.
  • encapsulated antibodies When encapsulated antibodies remain in the body for a long time, they may denature or aggregate as a result of exposure to moisture at 37°C., resulting in a loss of biological activity and possible changes in immunogenicity. Rational strategies can be devised for stabilization depending on the mechanism involved. For example, if the aggregation mechanism is discovered to be intermolecular S-S bond formation through thio-disulfide interchange, stabilization may be achieved by modifying sulihydryl residues, lyophilizing from acidic solutions, controlling moisture content, using appropriate additives, and developing specific polymer matrix compositions.
  • the polypeptide variant of the invention may be used as an affinity purification agent.
  • the polypeptide variant is immobilized on a solid phase such a Sephadex resin or filter paper, using methods well known in the art.
  • the immobilized polypeptide variant is contacted with a sample containing the antigen to be purified, and thereafter the support is washed with a suitable solvent that will remove substantially all the material in the sample except the antigen to be purified, which is bound to the immobilized polypeptide variant. Finally, the support is washed with another suitable solvent, such as glycine buffer, pH 5.0, that will release the antigen from the polypeptide variant.
  • polypeptide variant may also be useful in diagnostic assays, e.g., for detecting expression of an antigen of interest in specific cells, tissues, or serum.
  • polypeptide variant typically will be labeled with a detectable moiety.
  • labels are available which can be generally grouped into the following categories:
  • Radioisotopes such as 35 S, 14 C, 125 I, 3 H, and 131 I.
  • the polypeptide variant can be labeled with the radioisotope using the techniques described in Current Protocots in Immunology, Volumes 1 and 2, Coligen et al., Ed. Wiley-lnterscience, New York, New York, Pubs. (1991) for example and radioactivity can be measured using scintillation counting.
  • Fluorescent labels such as rare earth chelates (europium chelates) or fluorescein and its derivatives, rhodamine and its derivatives, dansyl, Lissamine, phycoerythrin and Texas Red are available.
  • the fluorescent labels can be conjugated to the polypeptide variant using the techniques disclosed in Current Protocols in Immunology, supra, for example. Fluorescence can be quantified using a fluorimeter.
  • the enzyme generally catalyzes a chemical alteration of the chromogenic substrate that can be measured using various techniques. For example, the enzyme may catalyze a color change in a substrate, which can be measured spectrophotometrically. Alternatively, the enzyme may alter the fluorescence or chemiluminescence of the substrate. Techniques for quantifying a change in fluorescence are described above.
  • the chemiluminescent substrate becomes electronically excited by a chemical reaction and may then emit light which can be measured (using a chemiluminometer, for example) or donates energy to a fluorescent acceptor.
  • enzymatic labels include luciferases (e.g., firefly luciferase and bacterial luciferase; U.S. Pat. No. 4,737,456), luciferin, 2,3-dihydrophthalazinediones, malate dehydrogenase, urease, peroxidase such as horseradish peroxidase (HRPO), alkaline phosphatase, ⁇ -galactosidase, glucoamylase, lysozyme, saccharide oxidases (e.g., glucose oxidase, galactose oxidase, and glucose-6-phosphate dehydrogenase), heterocyclic oxidases (such as uricase and xanthine oxidase), lactoperoxidase, microperoxidase, and the like.
  • luciferases e.g., firefly luciferase and bacterial lucifera
  • enzyme-substrate combinations include, for example:
  • HRPO Horseradish peroxidase
  • OPD orthophenylene diamine
  • TMB 3,3′,5,5′-tetramethyl benzidine hydrochloride
  • ⁇ -D-galactosidase ( ⁇ -D-Gal) with a chromogenic substrate (e.g., p-nitrophenyl- ⁇ -D-galactosidase) or fluorogenic substrate 4-methylumbelliferyl- ⁇ -D-galactosidase.
  • a chromogenic substrate e.g., p-nitrophenyl- ⁇ -D-galactosidase
  • fluorogenic substrate 4-methylumbelliferyl- ⁇ -D-galactosidase
  • the label is indirectly conjugated with the polypeptide variant.
  • the polypeptide variant can be conjugated with biotin and any of the three broad categories of labels mentioned above can be conjugated with avidin, or vice versa. Biotin binds selectively to avidin and thus, the label can be conjugated with the polypeptide variant in this indirect manner.
  • the polypeptide variant is conjugated with a small hapten (e.g., digoxin) and one of the different types of labels mentioned above is conjugated with an anti-hapten polypeptide variant (e.g., anti-digoxin antibody).
  • an anti-hapten polypeptide variant e.g., anti-digoxin antibody
  • the polypeptide variant need not be labeled, and the presence thereof can be detected using a labeled antibody which binds to the polypeptide variant.
  • polypeptide variant of the present invention may be employed in any known assay method, such as competitive binding assays, direct and indirect sandwich assays, and immunoprecipitation assays. Zola, Monoclonal Antibodies:A Manualof Techniques , pp.147-158 (CRC Press, Inc. 1987).
  • the polypeptide variant may also be used for in vivo diagnostic assays.
  • the polypeptide variant is labeled with a radionuclide (such as 111 In, 99 Tc, 14 C, 131 I, 125 I, 3 H, 32 P or 35 S) so that the antigen or cells expressing it can be localized using immunoscintiography.
  • a radionuclide such as 111 In, 99 Tc, 14 C, 131 I, 125 I, 3 H, 32 P or 35 S
  • the polypeptide variant of the present invention may be used to treat a mammal e.g. a patient suffering from a disease or disorder who could benefit from administration of the polypeptide variant.
  • the conditions which can be treated with the polypeptide variant are many and include cancer (e.g. where the polypeptide variant binds the HER2 receptor or CD20); allergic conditions such as asthma (with an anti-IgE antibody); and LFA-mediated disorders (e.g. where the polypeptide variant is an anti-LFA-1 or anti-ICAM-1 antibody) etc.
  • exemplary diseases or disorders to be treated include: cancer (e.g.
  • ADCC function is desirable, but complement activation would lead to amplified side effects, such as vasculitis in the blood vessels at the tumor site); disorders treated with an agonist antibody; disorders wherein the polypeptide variant binds a soluble antigen and wherein stoichiometry leads to immune complexes which activate the complement cascade and result in unwanted side effects; conditions employing an antagonist antibody which downmodulates receptor function without damaging tissue or organ function;
  • the polypeptide variant is administered by any suitable means, including parenteral, subcutaneous, intraperitoneal, intrapulmonary, and intranasal, and, if desired for local immunosuppressive treatment, intralesional administration.
  • Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.
  • the polypeptide variant is suitably administered by pulse infusion, particularly with declining doses of the polypeptide variant.
  • the dosing is given by injections, most preferably intravenous or subcutaneous injections, depending in part on whether the administration is brief or chronic.
  • polypeptide variant For the prevention or treatment of disease, the appropriate dosage of polypeptide variant will depend on the type of disease to be treated, the severity and course of the disease, whether the polypeptide variant is administered for preventive or therapeutic purposes, previous therapy, the patient's clinical history and response to the polypeptide variant, and the discretion of the attending physician.
  • the polypeptide variant is suitably administered to the patient at one time or over a series of treatments.
  • polypeptide variant is an initial candidate dosage for administration to the patient, whether, for example, by one or more separate administrations, or by continuous infusion.
  • a typical daily dosage might range from about 1 ⁇ g/kg to 100 mg/kg or more, depending on the factors mentioned above.
  • the treatment is sustained until a desired suppression of disease symptoms occurs.
  • other dosage regimens may be useful. The progress of this therapy is easily monitored by conventional techniques and assays.
  • the polypeptide variant composition will be formulated, dosed, and administered in a fashion consistent with good medical practice. Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners.
  • the “therapeutically effective amount” of the polypeptide variant to be administered will be governed by such considerations, and is the minimum amount necessary to prevent, ameliorate, or treat a disease or disorder.
  • the polypeptide variant need not be, but is optionally formulated with one or more agents currently used to prevent or treat the disorder in question.
  • the effective amount of such other agents depends on the amount of polypeptide variant present in the formulation, the type of disorder or treatment, and other factors discussed above. These are generally used in the same dosages and with administration routes as used hereinbefore or about from 1 to 99% of the heretofore employed dosages.
  • This assay determines binding of an IgG Fc region to recombinant Fc ⁇ RIIa, Fc ⁇ RIIb and Fc ⁇ RIIIa ⁇ subunits expressed as His6-glutathione S transferase (GST)-tagged fusion proteins. Since the affinity of the Fc region of IgG1 for the Fc ⁇ RI is in the nanomolar range, the binding of IgG1 Fc mutants can be measured by titrating monomeric IgG and measuring bound IgG with a polyclonal anti-lgG in a standard ELISA format (Example 2 below). The affinity of the other members of the Fc ⁇ R family, ie. Fc ⁇ RIIa, Fc ⁇ RIIb and Fc ⁇ RIIIa for IgG is however in the micromolar range and binding of monomeric IgG1 for these receptors can not be reliably measured in an ELISA format.
  • GST His6-glutathione S transferase
  • the following assay utilizes Fc mutants of recombinant anti-IgE E27 (FIGS. 4A and 4B) which, when mixed with human IgE at a 1:1 molar ratio, forms a stable hexamer consisting of three anti-IgE molecules and three IgE molecules.
  • a recombinant chimeric form of IgE (chimeric IgE) was engineered and consists of a human IgE Fc region and the Fab of an anti-VEGF antibody (Presta et al Cancer Research 57:4593-4599 (1997)) which binds two VEGF molecules per mole of anti-VEGF.
  • VEGF vascular endothelial growth factor
  • hexamers When recombinant human VEGF is added at a 2:1 molar ratio to chimeric IgE:E27 hexamers, the hexamers are linked into larger molecular weight complexes via the chimeric IgE Fab:VEGF interaction.
  • the E27 component of this complex binds to the Fc ⁇ RIIa, Fc ⁇ RIIb and Fc ⁇ IIIa ⁇ subunits with higher avidity to permit detection in an ELISA format.
  • Receptor Coat. Fc ⁇ receptor ⁇ subunits were expressed as GST fusions of His6 tagged extracellular domains (ECDs) in 293 cells resulting in an ECD-6His-GST fusion protein (Graham et al J. Gen. Virol . 36:59-72 (1977) and Gorman et al DNA Prot. Eng. Tech . 2:3-10 (1990)) and purified by Ni-NTA column chromatography (Qiagen, Australia) and buffer exchanged into phosphate buffered saline (PBS). Concentrations were determined by absorption at 280nm using extinction coefficients derived by amino acid composition analysis. Receptors were coated onto Nunc F96 maxisorb plates (cat no.
  • E27:chimeric IgE:VEGF(1:1:2 molar ratio) complexes are added to Fc ⁇ R ⁇ subunit coated plates at E27 concentrations of 5 ⁇ g and 1 ⁇ g total IgG in quadruplicate in assay buffer and incubated for 120 minutes at 25° C. on an orbital shaker.
  • HRP horse radish peroxidase conjugated goat anti-human IgG ( ⁇ ) heavy chain specific
  • Plates are washed 5 ⁇ with wash buffer to remove unbound HRP goat anti-human IgG and bound anti-lgG is detected by adding 100 ⁇ l of substrate solution (0.4mg/ml o-phenylenedaimine dihydrochloride, Sigma P6912, 6 mM H 2 O 2 in PBS) and incubating for 8 min at 25° C. Enzymatic reaction is stopped by the addition of 100 ⁇ l 4.5N H 2 SO 4 and colorimetric product is measured at 490 nm on a 96 well plate densitometer (Molecular Devices). Binding of E27 mutant complexes is expressed as a percent of the wild type E27 containing complex.
  • the heavy and light chain plasmids were co-transfected into an adenovirus transformed human embryonic kidney cell line as previously described (Werther et al., J. Immunol 157:4986 (1996)). The media was changed to serum-free 24 hours after transfection and the secreted antibody was harvested after five days.
  • the antibodies were purified using Protein A-SEPHAROSE CL-4BTM (Pharmacia), buffer exchanged and concentrated to 0.5 ml with PBS using a Centricon-30 (Amicon), and stored at 4° C. The concentration of the antibody was determined using total Ig-binding ELISA.
  • C1q Binding ELISA Costar 96 well plates were coated overnight at 4° C. with the indicated concentrations of 2B8 in coating buffer (0.05 M sodium carbonate buffer), pH 9. The plates were then washed 3 ⁇ with PBS/ 0.05% TWEEN 20TM, pH 7.4 and blocked with 200 ⁇ l of ELISA diluent without thimerosal (0.1 M NaPO4/0.1M NaCl/0.1% gelatin/0.05% TWEEN 20 TM/ 0.05% ProClin300) for 1 hr at room temperature.
  • the plate was washed 3 ⁇ with wash buffer, an aliquot of 100 ⁇ l of 2 ⁇ g/ml C1q (Quidel, San Diego, Calif.) was added to each well and incubated for 2 hrs at room temperature. The plate was then washed 6 ⁇ with wash buffer. 100 ⁇ l of a 1:1000 dilution of sheep anti-complement C1q peroxidase conjugated antibody (Biodesign) was added to each well and incubated for 1 hour at room temperature. The plate was again washed 6 ⁇ with wash buffer and 100 ⁇ l of substrate buffer (PBS/0.012% H 2 O 2 ) containing OPD (O-phenylenediamine dihydrochloride (Sigma)) was added to each well.
  • substrate buffer PBS/0.012% H 2 O 2
  • OPD O-phenylenediamine dihydrochloride
  • the oxidation reaction observed by the appearance of a yellow color, was allowed to proceed for 30 minutes and stopped by the addition of 100 ⁇ l of 4.5 N H 2 SO 4 .
  • the absorbance was then read at (492-405) nm using a microplate reader (SPECTRA MAX 250TM, Molecular Devices Corp.).
  • the appropriate controls were run in parallel (i.e. the ELISA was performed without C1q for each concentration of C2B8 used and also the ELISA was performed without C2B8).
  • C1q binding was measured by plotting the absorbance (492-405) nm versus concentration of C2B8 in ⁇ g/ml using a 4-parameter curve fitting program (KALEIDAGRAPHTM) and comparing EC 50 values.
  • Human complement (Quidel) was diluted 1:3 in RHB buffer and WIL2-S cells (available from the ATCC, Manassas, Va.) which express the CD20 antigen were diluted to a density of 1 ⁇ 10 6 cells /ml with RHB buffer.
  • Mixtures of 150 ⁇ l containing equal volumes of C2B8, diluted human complement and WIL2-S cells were added to a flat bottom tissue culture 96 well plate and allowed to incubate for 2 hrs at 37° C. and 5% CO 2 to facilitate complement mediated cell lysis. 50 ⁇ l of alamar blue (Accumed International) was then added to each well and incubated overnight at 37° C.
  • the absorbance was measured using a 96-well fluorometer with excitation at 530 nm and emission at 590 nm. As described by Gazzano-Santoro et al., the results are expressed in relative fluorescence units (RFU). The sample concentrations were computed from a C2B8 standard curve and the percent activity as compared to wild type C2B8 is reported for each mutant.
  • CD20 Binding Potency of the C2B8 Mutants The binding of C2B8 and mutants to the CD20 antigen were assessed by a method previously described (Reff et al, (1994), supra; reviewed in Gazzano-Santoro et al, (1996), supra). WIL2-S cells were grown for 3-4 days to a cell density of 1 ⁇ 10 6 cells/ml. The cells were washed and spun twice in FACS buffer (PBS/0.1% BSA/0.02% NaN 3 ) and resuspended to a cell density of 5 ⁇ 10 6 cells/ml.
  • FACS buffer PBS/0.1% BSA/0.02% NaN 3
  • Fc ⁇ RI ⁇ subunit-GST fusion was coated onto Nunc F96 maxisorb plates (cat no. 439454) by adding 100 ⁇ l of receptor-GST fusion at 1 ⁇ g/ml in PBS and incubated for 48 hours at 4° C. Prior to assay, plates are washed 3 ⁇ with 250 ⁇ l of wash buffer (PBS pH 7.4 containing 0.5% TWEEN 20TM) and blocked with 250 ⁇ l of assay buffer (50 mM Tris buffered saline, 0.05% TWEEN 20TM, 0.5% RIA grade bovine albumin (Sigma A7888), and 2 mM EDTA pH 7.4).
  • wash buffer PBS pH 7.4 containing 0.5% TWEEN 20TM
  • assay buffer 50 mM Tris buffered saline, 0.05% TWEEN 20TM, 0.5% RIA grade bovine albumin (Sigma A7888), and 2 mM EDTA pH 7.4
  • Plates are washed 5 ⁇ with wash buffer to remove unbound HRP goat anti-human IgG and bound anti-lgG is detected by adding 100 ⁇ l of substrate solution (0.4mg/ml o-phenylenedaimine dihydrochloride, Sigma P6912, 6 mM H 2 O 2 in PBS) and incubating for 8 min at 25° C. Enzymatic reaction is stopped by the addition of 100 ⁇ l 4.5N H 2 SO 4 and colorimetric product is measured at 490 nm on a 96 well plate densitometer (Molecular Devices). Binding of variant is expressed as a percent of the wild type molecule.
  • Fc ⁇ RII and III binding ELISAs were performed as described in Example 1 above.
  • the IgG2 mutant appears to have a much lower affinity for C1q than the E318A and K320A mutants (FIG. 2).
  • the results demonstrate that E318 and K320 do not constitute the core C1q binding sites for human IgG1.
  • the K322A substitution had a significant effect on both complement activity and C1q binding.
  • the K322A mutant did not have CDC activity when tested in the above CDC assay and was more than a 100 fold lower than wild type C2B8 in binding to C1q (FIG. 2).
  • K322 is the only residue of the proposed core C1q binding sites that appeared to have a significant effect on complement activation and C1q binding.
  • the mutation P329A results in an antibody that does not activate complement and is more than a 100 fold lower in binding to C1q (FIG. 3) when compared to wild type C2B8 (FIG. 2).

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Abstract

A variant of a polypeptide comprising a human IgG Fc region is described, which variant comprises an amino acid substitution at amino acid position 329, or at two or all of amino acid positions 329, 331 and 322 of the human IgG Fc region. Such variants display altered effector function. For example, Clq binding and/or complement dependent cytotoxicity (CDC) activity may be reduced or abolished in the variant polypeptide. The application also describes an immune complex and an assay for determining binding of an analyte, such as an Fc region-containing polypeptide, to a receptor.

Description

  • This is a non-provisional application claiming priority under 35 USC §119 to provisional application No. 60/080,447 filed Apr. 2, 1998, the entire disclosure of which is hereby incorporated by reference.[0001]
    • BACKGROUND OF THE INVENTION
  • 1. Field of the Invention [0002]
  • The present invention concerns variants of polypeptides comprising an Fc region. More particularly, the present invention concerns Fc region-containing polypeptides that have altered effector function as a consequence of one or more amino acid substitutions in the Fc region of the nonvariant polypeptide. The invention also relates to novel immune complexes and an assay for determining binding of an analyte, such as an Fc region-containing polypeptide, to a receptor. [0003]
  • 2. Description of Related Art [0004]
  • Antibodies are proteins, which exhibit binding specificity to a specific antigen. Native antibodies are usually heterotetrameric glycoproteins of about 150,000 daltons, composed of two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide linkages varies between the heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain has at one end a variable domain (V[0005] H) followed by a number of constant domains. Each light chain has a variable domain at one end (VL) and a constant domain at its other end; the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light chain variable domain is aligned with the variable domain of the heavy chain. Particular amino acid residues are believed to form an interface between the light and heavy chain variable domains.
  • The term “variable” refers to the fact that certain portions of the variable domains differ extensively in sequence among antibodies and are responsible for the binding specificity of each particular antibody for its particular antigen. However, the variability is not evenly distributed through the variable domains of antibodies. It is concentrated in three segments called complementarity determining regions (CDRs) both in the light chain and the heavy chain variable domains. The more highly conserved portions of the variable domains are called the framework regions (FR). The variable domains of native heavy and light chains each comprise four FR regions, largely adopting a βsheet configuration, connected by three CDRs, which form loops connecting, and in some cases forming part of, the β-sheet structure. The CDRs in each chain are held together in close proximity by the FR regions and, with the CDRs from the other chain, contribute to the formation of the antigen binding site of antibodies (see Kabat et al., Sequences of [0006] Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)).
  • The constant domains are not involved directly in binding an antibody to an antigen, but exhibit various effector functions. Depending on the amino acid sequence of the constant region of their heavy chains, antibodies or immunoglobulins can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG and IgM, and several of these may be further divided into subclasses (isotypes), e.g. IgG1, IgG2, IgG3, and IgG4; IgA1 and IgA2. The heavy chain constant regions that correspond to the different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively. Of the various human immunoglobulin classes, only human IgG1, IgG2, IgG3 and IgM are known to activate complement. [0007]
  • A schematic representation of the native IgG1 structure is shown in FIG. 1, where the various portions of the native antibody molecule are indicated. Papain digestion of antibodies produces two identical antigen binding fragments, called Fab fragments, each with a single antigen binding site, and a residual “Fc” fragment, whose name reflects its ability to crystallize readily. The crystal structure of the human IgG Fc region has been determined (Deisenhofer, [0008] Biochemistry 20:2361-2370 (1981)). In human IgG molecules the Fc region is generated by papain cleavage N-terminal to Cys 226. The Fc region is central to the effector functions of antibodies.
  • The effector functions mediated by the antibody Fc region can be divided into two categories: (1) effector functions that operate after the binding of antibody to an antigen (these functions involve the participation of the complement cascade or Fc receptor (FcR)-bearing cells); and (2) effector functions that operate independently of antigen binding (these functions confer persistence in the circulation and the ability to be transferred across cellular barriers by transcytosis). Ward and Ghetie, [0009] Therapeutic Immunology 2:77-94 (1995).
  • While binding of an antibody to the requisite antigen has a neutralizing effect that might prevent the binding of a foreign antigen to its endogenous target (e.g. receptor or ligand), binding alone may not remove the foreign antigen. To be efficient in removing and/or destructing foreign antigens, an antibody should be endowed with both high affinity binding to its antigen, and efficient effector functions. [0010]
    • C1q Binding
  • C1q and two serine proteases, C1r and C1s, form the complex C1, the first component of the complement dependent cytotoxicity (CDC) pathway. C1q is a hexavalent molecule with a molecular weight of approximately 460,000 and a structure likened to a bouquet of tulips in which six collagenous “stalks” are connected to six globular head regions. Burton and Woof, [0011] Advances in Immunol. 51:1-84 (1992). To activate the complement cascade, it is necessary for C1q to bind to at least two molecules of IgG1, IgG2, or IgG3 (the consensus is that IgG4 does not activate complement), but only one molecule of IgM, attached to the antigenic target. Ward and Ghetie, Therapeutic Immunology 2:77-94 (1995) at page 80.
  • Based upon the results of chemical modifications and crystallographic studies, Burton et aL ([0012] Nature, 288:338-344 (1980)) proposed that the binding site for the complement subcomponent C1q on IgG involves the last two (C-terminal) β-strands of the CH2 domain. Burton later suggested (Molec. Immunol., 22(3):161-206 (1985)) that the region comprising amino acid residues 318 to 337 might be involved in complement fixation.
  • Duncan and Winter ([0013] Nature 332:738-40 (1988)), using site directed mutagenesis, reported that Glu318, Lys320 and Lys322 form the binding site to C1q. The data of Duncan and Winter were generated by testing the binding of a mouse IgG2b isotype to guinea pig C1q. The role of Glu318, Lys320 and Lys322 residues in the binding of C1q was confirmed by the ability of a short synthetic peptide containing these residues to inhibit complement mediated lysis. Similar results are disclosed in U.S. Pat. No. 5,648,260 issued on Jul. 15, 1997, and U.S. Pat. No. 5,624,821 issued on Apr. 29, 1997.
  • The residue Pro331 has been implicated in C1q binding by analysis of the ability of human IgG subclasses to carry out complement mediated cell lysis. Mutation of Ser331 to Pro331 in IgG4 conferred the ability to activate complement. (Tao et aL, J. [0014] Exp. Med., 178:661-667 (1993); Brekke et al., Eur. J. lmmunol, 24:2542-47 (1994)).
  • From the comparison of the data of the Winter group, and the Tao et al. and Brekke et al. papers, Ward and Ghetie concluded in their review article that there are at least two different regions involved in the binding of C1q: one on the β-strand of the CH2 domain bearing the Glu318, Lys320 and Lys322 residues, and the other on a turn located in close proximity to the sameβ-strand, and containing a key amino acid residue at position 331. [0015]
  • Other reports suggested that human IgG1 residues Lys235, and Gly237, located in the lower hinge region, play a critical role in complement fixation and activation. Xu et al., [0016] J. Immunol. 150:152A (Abstract) (1993). WO94/29351 published Dec. 22, 1994 reports that amino acid residues necessary for C1q and FcR binding of human IgG1 are located in the N-terminal region of the CH2 domain, i.e. residues 231 to 238.
  • It has further been proposed that the ability of IgG to bind C1q and activate the complement cascade also depends on the presence, absence or modification of the carbohydrate moiety positioned between the two CH2 domains (which is normally anchored at Asn297). Ward and Ghetie, [0017] Therapeutic Immunology 2:77-94 (1995) at page 81.
    • Fc Receptor Binding
  • Effector functions can also be mediated by the interaction of the Fc region of an antibody with Fc receptors (FcRs), which are specialized cell surface receptors on hematopoietic cells. Fc receptors belong in the immunoglobulin superfamily, and have been shown to mediate both the removal of antibody-coated pathogens by phagocytosis of immune complexes, and the lysing of erythrocytes and various other cellular targets (e.g. tumor cells) coated with the corresponding antibody, via antibody dependent cell mediated cytotoxicity (ADCC). Van de Winkel and Anderson, [0018] J. Leuk. Biol. 49:511-24 (1991).
  • FcRs are defined by their specificity for immunoglobulin isotypes; Fc receptors for IgG antibodies are referred to as FcγR, for IgE as FcεR, for IgA as FcαR and so on. Three subclasses of gamma receptors have been identified: FcεRI (CD64), FcγRII (CD32) and FcγRIII (CD16). Because each FcγR subclass is encoded by two or three genes, and alternative RNA spicing leads to multiple transcripts, a broad diversity in FcγR isoforms exists. The three genes encoding the FcγRI subclass (FcγRIA, FcγRIB and FcγRIC) are clustered in region 1q21.1 of the long arm of [0019] chromosome 1; the genes encoding FcγRII isoforms (FcγRIIA, FcγRIIB and FcγRIIC) and the two genes encoding FcγRIII (FcγRIIIA and FcγRIIIB) are all clustered in region 1q22. FcRs are reviewed in Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991); Capel et aL, Immunomethods 4:25-34 (1994); and de Haas etal., J. Lab. Clin. Med. 126:330-41 (1995).
  • While FcγRI binds monomeric IgG with a high affinity, FcγRII and FcγRIII are low-affinity receptors, interacting with complexed or aggregated IgG. The classical method for detecting these low-affinity receptors is by “rosetting” using antibody-coated erythrocytes (EA) sensitized with IgGs. Bredius et aL evaluated rosette formation between IgG-sensitized red blood cells and polymorphonuclear leukocytes (PMN) which express FcγRIIa and FcγRIIIb at their cell-surface. Rosette was defined as three of more EA bound per PMN (Bredius etal. [0020] Immunology 83:624-630 (1994)). See, also, Tax et aL J. Immunol. 133(3):1185-1189 (1984); Nagarajan et aL J. Biol. Chem. 270(43):25762-25770 (1995); and Warmerdam et al. J. Immunol. 147(4):1338-1343 (1991concerning rosette assays. However, binding of these EA “immune complexes” to FcR is not easily quantified. Accordingly, more defined complexes with detectable affinity for these FcRs have been developed. For example, IgG dimers have been formed using anti-light chain monoclonal antibodies (Nagarajan et al, supra and Warmerdam et al., supra) or chemical cross-linking agents (Hogarth et al. Immunomethods 4:17-24 (1994); and Tamm et aL J. Biol Chem. 271(7):3659-3666(1996)). Heat-aggregated immune complexes have also been evaluated for binding to cells expressing FcRs (Tax et al., supra and Tam et al., supra).
  • The binding site for the FcγRs on human IgG was found to reside in the lower hinge region, primarily involving residues at amino acid positions 233-238, all of which were found to be necessary for full FcγR binding activity. Canfield and Morrison, [0021] J. Exp. Med. 173:1483-91 (1991); Chappel et al., Proc. Natl. Acad. Sci. USA, 88:9036-40 (1991); Lund etal., J. Immunol., 147:2657-62 (1991); Lund et al, Molec. ImmunoL., 29:53-59 (1992); Jefferis et al., Molec. Immunol., 27:1237-40 (1990); and Sarmay et al., Molec. Immunol, 29:633-639 (1992).
  • Pro331 in IgG3 was changed to Ser, and the affinity of this mutant to target cells analyzed. The affinity was found to be six-fold lower than that of unmutated IgG3, indicating the involvement of Pro331 in FcγRI binding. Morrison et al., [0022] Immunologist, 2:119-124 (1994); and Canfield and Morrison, J. Exp. Med. 173:1483-91 (1991).
  • In addition, Glu318 was identified as being involved in binding to FcγRII. Ward and Ghetie, [0023] Therapeutic Immunology 2:77-94 (1995).
    • SUMMARY OF THE INVENTION
  • The present invention provides a variant of a polypeptide comprising a human IgG Fc region, which variant comprises an amino acid substitution at amino acid position 329, or at two or all of amino acid positions 329, 331 and 322 of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat. [0024]
  • The invention further relates to a variant of a polypeptide comprising a human IgG Fc region, which variant binds FcγRI, FcγRII, FcγRIII and FcRn but does not activate complement and comprises an amino acid substitution at amino acid position 322 or amino acid position 329, or both amino acid positions of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat. [0025]
  • In yet a further aspect, the invention provides a method for modifying a polypeptide comprising a human IgG Fc region comprising substituting an amino acid residue at amino acid position 329, or at two or all of amino acid positions 329, 331 and 322 of the human lgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat. [0026]
  • The invention also provides a composition comprising the polypeptide variant and a physiologically acceptable carrier or diluent. This composition for potential therapeutic use is sterile and may be lyophilized. [0027]
  • Diagnostic and therapeutic uses for the polypeptide variant are contemplated. In one diagnostic application, the invention provides a method for determining the presence of a protein of interest comprising exposing a sample suspected of containing the protein to the polypeptide variant and determining binding of the polypeptide variant to the sample. In one therapeutic application, the invention provides a method of treating a mammal suffering from a disorder comprising administering to the mammal a therapeutically effective amount of a variant of a polypeptide comprising a human IgG Fc region, which variant binds FcγRI, FcγRII, FcγRIII and FcRn but does not activate complement and comprises an amino acid substitution at amino acid position 329, 331 or 322 of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat. [0028]
  • The invention further provides: isolated nucleic acid encoding the polypeptide variant; a vector comprising the nucleic acid, optionally, operably linked to control sequences recognized by a host cell transformed with the vector; a host cell comprising the vector; a process for producing the polypeptide variant comprising culturing this host cell so that the nucleic acid is expressed and, optionally, recovering the polypeptide variant from the host cell culture (e.g. from the host cell culture medium). [0029]
  • The invention also pertains to an immune complex comprising: (a) an Fc region-containing polypeptide; (b) a first target molecule which comprises at least two binding sites for the Fc region-containing polypeptide; and (c) a second target molecule comprises at least two binding sites for the first target molecule. The immune complex may be used in an FcR-binding assay, particularly where the FcR has a low affinity for the Fc region-containing polypeptide. Other uses for the immune complex are disclosed herein. [0030]
  • Moreover, the invention provides a method for determining binding of an analyte, such as an Fc region-containing polypeptide, to a receptor (e.g. a low affinity FcR) comprising the following steps performed sequentially: (a) forming a molecular complex between the analyte and a first target molecule, wherein the first target molecule comprises at least two binding sites for the analyte; and (b) determining binding of the molecular complex of step (a) to the receptor (e.g. to a binding domain of the receptor coated on an assay plate). Optionally, the molecular complex of step (a) further comprises a second target molecule which comprises at least two binding sites for the first target molecule. [0031]
  • The invention also relates to an assay kit, such as a kit useful for determining binding of an analyte to a receptor comprising: (a) a first target molecule which comprises at least two binding sites for the analyte; and (b) a second target molecule which comprises at least two binding sites for the first target molecule.[0032]
    • BRIEF DESCRIPTIOM OF THE DRAWINDS
  • FIG. 1 is a schematic representation of a native IgG. Disulfide bonds are represented by heavy lines between CH1 and CL domains and the two CH2 domains. V is variable domain; C is constant domain; L stands for light chain and H stands for heavy chain. [0033]
  • FIG. 2 shows C1q binding of wild type (wt) C2B8 antibody; C2B8 antibody with a human IgG2 constant region (IgG2); and mutants K322A, K320A and E318A. [0034]
  • FIG. 3 depicts C1q binding of mutants P331A, P329A and K322A. [0035]
  • FIGS. 4A (SEQ ID NO:1) and 4B (SEQ ID NO:2) depict the amino acid sequences of E27 anti-IgE antibody light chain (FIG. 4A) and heavy chain (FIG. 4B). [0036]
  • FIG. 5 is a schematic diagram of the “immune complex” prepared for use in the FcR assay described in Example 1. The hexamer comprising three anti-IgE antibody molecules (the “Fc region-containing polypeptide”) and three IgE molecules (the “first target molecule”) is shown. IgE has two “binding sites” for the anti-lgE antibody (E27) in the Fc region thereof. Each IgE molecule in the complex is further able to bind two VEGF molecules (“the second target polypeptide”). VEGF has two “binding sites” for IgE.[0037]
    • DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
  • I. Definitions [0038]
  • Throughout the present specification and claims, the numbering of the residues in an IgG heavy chain is that of the EU index as in Kabat et al., [0039] Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991), expressly incorporated herein by reference. The “EU index as in Kabat” refers to the residue numbering of the human IgG1 EU antibody.
  • The term “Fc region” is used to define a C-terminal region of an IgG heavy chain as shown FIG. 1. Although the boundaries of the Fc region of an IgG heavy chain might vary slightly, the human IgG heavy chain Fc region is usually defined to stretch from amino acid residue at position Cys226 to the carboxyl-terminus. The term “Fc region-containing polypeptide” refers to a polypeptide, such as an antibody or immunoadhesin (see definitions below), which comprises an Fc region. [0040]
  • The Fc region of an IgG comprises two constant domains, CH2 and CH3, as shown in FIG. 1. The “CH2” domain of a human IgG Fc region (also referred to as “Cγ2” domain) usually extends from amino acid 231 to amino acid 340. The CH2 domain is unique in that it is not closely paired with another domain. Rather, two N-linked branched carbohydrate chains are interposed between the two CH2 domains of an intact native IgG molecule. It has been speculated that the carbohydrate may provide a substitute for the domain-domain pairing and help stabilize the CH2 domain. Burton, [0041] Molec. Immunol. 22:161-206 (1985).
  • “Hinge region” is generally defined as stretching from Glu216 to Pro230 of human IgG1 (Burton, [0042] Molec. Immunol. 22:161-206 (1985)) Hinge regions of other IgG isotypes may be aligned with the IgG1 sequence by placing the first and last cysteine residues forming inter-heavy chain S-S bonds in the same positions.
  • “C1q” is a polypeptide that includes a binding site for the Fc region of an immunoglobulin. C1q together with two serine proteases, C1r and C1s, forms the complex C1, the first component of the complement dependent cytotoxicity (CDC) pathway. Human C1q can be purchased commercially from, e.g. Quidel, San Diego, CA. [0043]
  • The term “Fc receptor” or “FcR” is used to describe a receptor that binds to the Fc region of an antibody. The preferred FcR is one, which binds an IgG antibody (a gamma receptor) and includes receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and alternatively spliced forms of these receptors. FcRs are reviewed in Ravetch and Kinet, [0044] Annu. Rev. Immunol 9:457-92 (1991); Capel et al, Immunomethods 4:25-34 (1994); and de Haas et al, J. Lab. Clin. Med. 126:330-41 (1995). Other FcRs, including those to be identified in the future, are encompassed by the term “FcR” herein. The term also includes the neonatal receptor, FcRn, which is responsible for the transfer of maternal IgGs to the fetus (Guyer et al., J. Immunol. 117:587 (1976) and Kim et al., J. Immunol. 24:249 (1994)).
  • The term “binding domain” refers to the region of a polypeptide that binds to another molecule. In the case of an FcR, the binding domain can comprise a portion of a polypeptide chain thereof (e.g. the α chain thereof) which is responsible for binding an Fc region. One useful binding domain is the extracellular domain of an FcR α chain. [0045]
  • The term “antibody” is used in the broadest sense and specifically covers monoclonal antibodies (including full length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments so long as they exhibit the desired biological activity. [0046]
  • “Antibody fragments”, as defined for the purpose of the present invention, comprise a portion of an intact antibody, generally including the antigen binding or variable region of the intact antibody, so long as they retain at least the CH2 region of an IgG immunoglobulin heavy chain constant domain, comprising amino acid residues 322, 329 and 331, and have the ability, alone or in combination with another antibody fragment, to specifically bind a selected antigen. Examples of antibody fragments include linear antibodies; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments. The antibody fragments preferably retain at least part of the hinge and optionally the CH1 region of an IgG heavy chain. More preferably, the antibody fragments retain the entire constant region of an IgG heavy chain, and include an IgG light chain. [0047]
  • The term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to conventional (polyclonal) antibody preparations that typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. The modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method first described by Kohler et al, [0048] Nature 256:495 (1975), or may be made by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567). The “monoclonal antibodies” may also be isolated from phage antibody libraries using the techniques described in Clackson et al., Nature 352:624-628 (1991) and Marks et al., J. Mol. Biol. 222:581-597 (1991), for example.
  • The monoclonal antibodies herein specifically include “chimeric” antibodies (immunoglobulins) in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (U.S. Pat. No. 4,816,567; and Morrison et al, [0049] Proc. Natl. Acad. Sci. USA 81:6851-6855 (1984)).
  • “Humanized” forms of non-human (e.g., murine) antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulin. For the most part, humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a hypervariable region of the recipient are replaced by residues from a hypervariable region of a non-human species (donor antibody) such as mouse, rat, rabbit or nonhuman primate having the desired specificity, affinity, and capacity. In some instances, Fv framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues that are not found in the recipient antibody or in the donor antibody. These modifications are made to further refine antibody performance. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin sequence. The humanized antibody optionally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see Jones et al., [0050] Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992).
  • The term “hypervariable region” when used herein refers to the amino acid residues of an antibody which are responsible for antigen-binding. The hypervariable region comprises amino acid residues from a “complementarity determining region” or “CDR” (i.e. residues 24-34 (L1), 50-56 (L2) and 89-97 (L3) in the light chain variable domain and 31-35 (H1), 50-65 (H2) and 95-102 (H3) in the heavy chain variable domain; Kabat et al., [0051] Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)) and/or those residues from a “hypervariable loop” (i.e. residues 26-32 (L1), 50-52 (L2) and 91-96 (L3) in the light chain variable domain and 26-32 (H1), 53-55 (H2) and 96-101 (H3) in the heavy chain variable domain; Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). “Framework” or “FR” residues are those variable domain residues other than the hypervariable region residues as herein defined.
  • As used herein, the term “immunoadhesin” designates antibody-like molecules which combine the “binding domains” of a heterologous “adhesin” protein (e.g. a receptor, ligand or enzyme) with an immunoglobulin constant domain. Structurally, the immunoadhesins comprise a fusion of the adhesin amino acid sequence with the desired binding specificity which is other than the antigen recognition and binding site (antigen combining site) of an antibody (i.e. is “heterologous”) and an immunoglobulin constant domain sequence. [0052]
  • The term “ligand binding domain” as used herein refers to any native cell-surface receptor or any region or derivative thereof retaining at least a qualitative ligand binding ability of a corresponding native receptor. In a specific embodiment, the receptor is from a cell-surface polypeptide having an extracellular domain that is homologous to a member of the immunoglobulin supergenefamily. Other receptors, which are not members of the immunoglobulin supergenefamily but are nonetheless specifically covered by this definition, are receptors for cytokines, and in particular receptors with tyrosine kinase activity (receptor tyrosine kinases), members of the hematopoietin and nerve growth factor receptor superfamilies, and cell adhesion molecules, e.g. (E-, L- and P-) selectins. [0053]
  • The term “receptor binding domain” is used to designate any native ligand for a receptor, including cell adhesion molecules, or any region or derivative of such native ligand retaining at least a qualitative receptor binding ability of a corresponding native ligand. This definition, among others, specifically includes binding sequences from ligands for the above-mentioned receptors. [0054]
  • An “antibody-immunoadhesin chimera” comprises a molecule that combines at least one binding domain of an antibody (as herein defined) with at least one immunoadhesin (as defined in this application). Exemplary antibody-immunoadhesin chimeras are the bispecific CD4-lgG chimeras described in Berg et al, [0055] PNAS (USA) 88:4723-4727 (1991) and Chamow et al, J. Immunol. 153:4268 (1994).
  • An “isolated” polypeptide is one that has been identified and separated and/or recovered from a component of its natural environment. Contaminant components of its natural environment are materials that would interfere with diagnostic or therapeutic uses for the polypeptide, and may include enzymes, hormones, and other proteinaceous or nonproteinaceous solutes. In preferred embodiments, the polypeptide will be purified (1) to greater than 95% by weight of polypeptide as determined by the Lowry method, and most preferably more than 99% by weight, (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence by use of a spinning cup sequenator, or (3) to homogeneity by SDS-PAGE under reducing or nonreducing conditions using Coomassie blue or, preferably, silver stain. Isolated polypeptide includes the polypeptide in situ within recombinant cells since at least one component of the polypeptide's natural environment will not be present. Ordinarily, however, isolated polypeptide will be prepared by at least one purification step. [0056]
  • “Treatment” refers to both therapeutic treatment and prophylactic or preventative measures. Those in need of treatment include those already with the disorder as well as those in which the disorder is to be prevented. [0057]
  • A “disorder” is any condition that would benefit from treatment with the polypeptide variant. This includes chronic and acute disorders or diseases including those pathological conditions which predispose the mammal to the disorder in question. [0058]
  • The word “label” when used herein refers to a detectable compound or composition which is conjugated directly or indirectly to the polypeptide. The label may be itself be detectable (e.g., radioisotope labels or fluorescent labels) or, in the case of an enzymatic label, may catalyze chemical alteration of a substrate compound or composition which is detectable. [0059]
  • An “isolated” nucleic acid molecule is a nucleic acid molecule that is identified and separated from at least one contaminant nucleic acid molecule with which it is ordinarily associated in the natural source of the polypeptide nucleic acid. An isolated nucleic acid molecule is other than in the form or setting in which it is found in nature. Isolated nucleic acid molecules therefore are distinguished from the nucleic acid molecule as it exists in natural cells. However, an isolated nucleic acid molecule includes a nucleic acid molecule contained in cells that ordinarily express the polypeptide where, for example, the nucleic acid molecule is in a chromosomal location different from that of natural cells. [0060]
  • The expression “control sequences” refers to DNA sequences necessary for the expression of an operably linked coding sequence in a particular host organism. The control sequences that are suitable for prokaryotes, for example, include a promoter, optionally an operator sequence, and a ribosome binding site. Eukaryotic cells are known to utilize promoters, polyadenylation signals, and enhancers. [0061]
  • Nucleic acid is “operably linked” when it is placed into a functional relationship with another nucleic acid sequence. For example, DNA for a presequence or secretory leader is operably linked to DNA for a polypeptide if it is expressed as a preprotein that participates in the secretion of the polypeptide; a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence; or a ribosome binding site is operably linked to a coding sequence if it is positioned so as to facilitate translation. Generally, “operably linked” means that the DNA sequences being linked are contiguous, and, in the case of a secretory leader, contiguous and in reading phase. However, enhancers do not have to be contiguous. Linking is accomplished by ligation at convenient restriction sites. If such sites do not exist, the synthetic oligonucleotide adaptors or linkers are used in accordance with conventional practice. [0062]
  • As used herein, the expressions “cell,” “cell line,” and “cell culture” are used interchangeably and all such designations include progeny. Thus, the words “transformants” and “transformed cells” include the primary subject cell and cultures derived therefrom without regard for the number of transfers. It is also understood that all progeny may not be precisely identical in DNA content, due to deliberate or inadvertent mutations. Mutant progeny that have the same function or biological activity as screened for in the originally transformed cell are included. Where distinct designations are intended, it will be clear from the context. [0063]
  • The term “molecular complex” when used herein refers to the relatively stable structure which forms when two or more heterologous molecules (e.g. polypeptides) bind (preferably noncovalently) to one another. The preferred molecular complex herein is an immune complex. “Immune complex” refers to the relatively stable structure which forms when at least one target molecule and at least one heterologous Fc region-containing polypeptide bind to one another forming a larger molecular weight complex. Examples of immune complexes are antigen-antibody aggregates and target molecule-immunoadhesin aggregates. The term “immune complex” as used herein, unless indicated otherwise, refers to an ex vivo complex (i.e. other than the form or setting in which it may be found in nature). However, the immune complex may be administered to a mammal, e.g. to evaluate clearance of the immune complex in the mammal. [0064]
  • The term “target molecule” refers to a molecule, usually a polypeptide, which is capable of being bound by a heterologous molecule and has one or more binding sites for the heterologous molecule. The term “binding site” refers to a region of a molecule to which another molecule can bind. The “first target molecule” herein comprises at least two distinct binding sites (for example, two to five separate binding sites) for an analyte (e.g. an Fc region-containing polypeptide) such that at least two analyte molecules can bind to the first target molecule. In the preferred embodiment of the invention, the two or more binding sites are identical (e.g. having the same amino acid sequence, where the target molecule is a polypeptide). In Example 1 below, the first target molecule was IgE and had two separate binding sites in the Fc region thereof to which the Fc region-containing polypeptide (an anti-IgE antibody, E27) could bind. Other first target molecules include dimers of substantially identical monomors (e.g. neurotrophins, IL8 and VEGF) or are polypeptides comprising two or more substantially identical polypeptide chains (e.g. antibodies or immunoadhesins). The “second target molecule” comprises at least two distinct binding sites (for example, two to five separate binding sites) for the first target molecule such that at least two first target molecules can bind to the second target molecule. Preferably, the two or more binding sites are identical (e.g. having the same amino acid sequence, where the target molecule is a polypeptide). In Example 2, the second target molecule was VEGF, which has a pair of distinct binding sites to which the variable domain of the IgE antibody could bind. Other second target molecules are contemplated, e.g. other dimers of substantially identical monomers (e.g. neurotrophins or IL8) or polypeptides comprising two or more substantially identical domains (e.g. antibodies or immunoadhesins). [0065]
  • An “analyte” is a substance that is to be analyzed. The preferred analyte is an Fc region-containing polypeptide that is to be analyzed for its ability to bind to an Fc receptor. [0066]
  • A “receptor” is a polypeptide capable of binding at least one ligand. The preferred receptor is a cell-surface receptor having an extracellular ligand-binding domain and, optionally, other domains (e.g. transmembrane domain, intracellular domain and/or membrane anchor). The receptor to be evaluated in the assay described herein may be an intact receptor or a fragment or derivative thereof (e.g. a fusion protein comprising the binding domain of the receptor fused to one or more heterologous polypeptides). Moreover, the receptor to be evaluated for its binding properties may be present in a cell or isolated and optionally coated on an assay plate or some other solid phase. [0067]
  • The phrase “low affinity receptor” denotes a receptor that has a weak binding affinity for a ligand of interest, e.g. having a binding constant of about 50 nM or worse affinity. Exemplary low affinity receptors include FcγRII and FcγRIII as well as adhesion molecules, such as integrins. [0068]
  • II. Modes for Carrying out the Invention [0069]
  • The invention herein relates to a method for making a polypeptide variant. The “starting” or “nonvariant” polypeptide is prepared using techniques available in the art for generating polypeptides comprising an Fc region. In the preferred embodiment of the invention, the polypeptide is an antibody and exemplary methods for generating antibodies are described in more detail in the following sections. The polypeptide may, however, be any other polypeptide comprising an Fc region, e.g. an immunoadhesin. Methods for making immunoadhesins are elaborated in more detail hereinbelow. [0070]
  • The starting polypeptide of particular interest herein is usually one that binds to C1q and displays complement dependent cytotoxicity (CDC). The amino acid substitutions described herein will generally serve to alter the ability of the starting polypeptide to bind to C1q and/or modify its complement dependent cytotoxicity function, e.g. to reduce and preferably abolish these effector functions. However, polypeptides comprising substitutions at one or more of the described positions with improved effector functions are contemplated herein. For example, the starting polypeptide may be unable to bind C1q and/or mediate CDC and may be modified according to the teachings herein such that it acquires these effector functions. Moreover, polypeptides with pre-existing C1q binding activity, optionally further having the ability to mediate CDC may be modified such that one or both of these activities are enhanced. [0071]
  • To generate the polypeptide variant, one or more amino acid alterations (e.g. substitutions) are introduced in the Fc region of the starting polypeptide. The amino acid positions to be modified are selected from heavy chain positions 329, 331 and 322, where the numbering of the residues in an IgG heavy chain is that of the EU index as in Kabat et al, [0072] Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md (1991). The Fc region is preferably from a human IgG and most preferably a human IgG1. The human IgG1 Fc region may be a human A or non-A allotype. Proline is conserved at position 329 in human IgG's. This residue is preferably replaced with alanine, however substitution with any other amino acid is contemplated, e.g., serine, threonine, asparagine, glycine or valine. Proline is conserved at position 331 in human IgG1, IgG2 and IgG3, but not IgG4 (which has a serine residue at position 331). Residue 331 is preferably replaced by alanine or another amino acid, e.g. serine (for IgG regions other than IgG4), glycine or valine. Lysine 322 is conserved in human IgGs, and this residue is preferably replaced by an alanine residue, but substitution with any other amino acid residue is contemplated, e.g. serine, threonine, glycine or valine.
  • In one embodiment, only one of the three above-identified positions is altered in order to generate the polypeptide variant. Preferably only residue 329 or only residue 322 is altered if this is the case. Alternatively, two or all of the three identified positions are modified. Thus, a variant may be generated in which the native amino acid residue at position 329 of the human heavy chain constant region is substituted with another amino acid, optionally in combination with a substitution of the amino acid residue at position 331 and/or substitution of the amino acid residue at position 322. Otherwise, the native amino acid residue at position 331 and the native amino acid residue at position 322 of the human IgG Fc region may both be substituted with another amino acid residue. [0073]
  • DNA encoding amino acid sequence variant of the starting polypeptide is prepared by a variety of methods known in the art. These methods include, but are not limited to, preparation by site-directed (or oligonucleotide-mediated) mutagenesis, PCR mutagenesis, and cassette mutagenesis of an earlier prepared DNA encoding the polypeptide [0074]
  • Site-directed mutagenesis is a preferred method for preparing substitution variants. This technique is well known in the art (see, e.g.,Carter et al. [0075] Nucleic Acids Res. 13:4431-4443 (1985) and Kunkel et al, Proc. Natl. Acad.Sci.USA 82:488 (1987)). Briefly, in carrying out site-directed mutagenesis of DNA, the starting DNA is altered by first hybridizing an oligonucleotide encoding the desired mutation to a single strand of such starting DNA. After hybridization, a DNA polymerase is used to synthesize an entire second strand, using the hybridized oligonucleotide as a primer, and using the single strand of the starting DNA as a template. Thus, the oligonucleotide encoding the desired mutation is incorporated in the resulting double-stranded DNA.
  • PCR mutagenesis is also suitable for making amino acid sequence variants of the starting polypeptide. See Higuchi, in [0076] PCR Protocols, pp. 177-183 (Academic Press, 1990); and Vallette et al., Nuc. Acids Res. 17:723-733 (1989). Briefly, when small amounts of template DNA are used as starting material in a PCR, primers that differ slightly in sequence from the corresponding region in a template DNA can be used to generate relatively large quantities of a specific DNA fragment that differs from the template sequence only at the positions where the primers differ from the template.
  • Another method for preparing variants, cassette mutagenesis, is based on the technique described by Wells et al., [0077] Gene 34:315-323 (1985). The starting material is the plasmid (or other vector) comprising the starting polypeptide DNA to be mutated. The codon(s) in the starting DNA to be mutated are identified. There must be a unique restriction endonuclease site on each side of the identified mutation site(s). If no such restriction sites exist, they may be generated using the above-described oligonucleotide-mediated mutagenesis method to introduce them at appropriate locations in the starting polypeptide DNA. The plasmid DNA is cut at these sites to linearize it. A double-stranded oligonucleotide encoding the sequence of the DNA between the restriction sites but containing the desired mutation(s) is synthesized using standard procedures, wherein the two strands of the oligonucleotide are synthesized separately and then hybridized together using standard techniques. This double-stranded oligonucleotide is referred to as the cassette. This cassette is designed to have 5′ and 3′ ends that are compatible with the ends of the linearized plasmid, such that it can be directly ligated to the plasmid. This plasmid now contains the mutated DNA sequence.
  • The polypeptide variant(s) so prepared may be subjected to further modifications, oftentimes depending on the intended use of the polypeptide. Such modifications may involve further alteration of the amino acid sequence, fusion to heterologous polypeptide(s) and/or covalent modifications. For example, it may be useful to combine the above amino acid substitutions with one or more further amino acid substitutions that reduce or ablate FcR binding. For example, the native amino acid residues at any one or more of heavy chain positions 233-238, 318 or 331 (where the numbering of the residues in an IgG heavy chain is that of the EU index as in Kabat et al, supra) may be replaced with non-native residue(s), e.g. alanine. [0078]
  • With respect to further amino acid sequence alterations, any cysteine residue not involved in maintaining the proper conformation of the polypeptide variant also may be substituted, generally with serine, to improve the oxidative stability of the molecule and prevent aberrant cross linking. [0079]
  • Another type of amino acid substitution serves to alter the glycosylation pattern of the polypeptide. This may be achieved by deleting one or more carbohydrate moieties found in the polypeptide, and/or adding one or more glycosylation sites that are not present in the polypeptide. Glycosylation of polypeptides is typically either N-linked or O-linked. N-linked refers to the attachment of the carbohydrate moiety to the side chain of an asparagine residue. The tripeptide sequences asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline, are the recognition sequences for enzymatic attachment of the carbohydrate moiety to the asparagine side chain. Thus, the presence of either of these tripeptide sequences in a polypeptide creates a potential glycosylation site. O-linked glycosylation refers to the attachment of one of the sugars N-aceylgalactosamine, galactose, or xylose to a hydroxyamino acid, most commonly serine or threonine, although 5-hydroxyproline or 5-hydroxylysine may also be used. Addition of glycosylation sites to the polypeptide is conveniently accomplished by altering the amino acid sequence such that it contains one or more of the above-described tripeptide sequences (for N-linked glycosylation sites). The alteration may also be made by the addition of, or substitution by, one or more serine or threonine residues to the sequence of the original polypeptide (for O-linked glycosylation sites). An exemplary glycosylation variant has an amino acid substitution of residue Asn 297 of the heavy chain. [0080]
  • The polypeptide variant may be subjected to one or more assays to evaluate any change in biological activity compared to the starting polypeptide. For example, the ability of the variant to bind C1q and mediate complement dependent cytotoxicity (CDC) may be assessed. [0081]
  • To determine C1q binding, a C1q binding ELISA may be performed. Briefly, assay plates may be coated overnight at 4° C. with polypeptide variant or starting polypeptide (control) in coating buffer. The plates may then be washed and blocked. Following washing, an aliquot of human C1q may be added to each well and incubated for 2 hrs at room temperature. Following a further wash, 100 μl of a sheep anti-complement C1q peroxidase conjugated antibody may be added to each well and incubated for 1 hour at room temperature. The plate may again be washed with wash buffer and 100 μl of substrate buffer containing OPD ( O-phenylenediamine dihydrochloride (Sigma)) may be added to each well. The oxidation reaction, observed by the appearance of a yellow color, may be allowed to proceed for 30 minutes and stopped by the addition of 100 μl of 4.5 N H[0082] 2SO4. The absorbance may then read at (492-405) nm.
  • The preferred polypeptide variant is one that displays a “significant reduction in C1q binding” in this assay. This means that about 100 μg/ml of the polypeptide variant displays about 50 fold or more reduction in C1q binding compared to 100 μg/ml of a control antibody having a nonmutated IgG1 Fc region. In the most preferred embodiment, the polypeptide variant “does not bind C1q”, i.e. 100 μg/ml of the polypeptide variant displays about 100 fold or more reduction in C1q binding compared to 100 μg/ml of the control antibody. [0083]
  • To assess complement activation, a complement dependent cytotoxicity (CDC) assay may be performed, e.g. as described in Gazzano-Santoro et al., [0084] J. Immunol. Methods 202:163 (1996). Briefly, various concentrations of the polypeptide variant and human complement may be diluted with buffer. Cells which express the antigen to which the polypeptide variant binds may be diluted to a density of˜1×106 cells/ ml. Mixtures of polypeptide variant, diluted human complement and cells expressing the antigen may be added to a flat bottom tissue culture 96 well plate and allowed to incubate for 2 hrs at 37°C. and 5% CO2 to facilitate complement mediated cell lysis. 50 μl of alamar blue (Accumed International) may then be added to each well and incubated overnight at 37°C. The absorbance is measured using a 96-well fluorometer with excitation at 530 nm and emission at 590 nm. The results may be expressed in relative fluorescence units (RFU). The sample concentrations may be computed from a standard curve and the percent activity as compared to nonvariant polypeptide is reported for the polypeptide variant of interest.
  • The preferred variant “does not activate complement”. For example, 0.6 μ/ml of the polypeptide variant displays about 0-10% CDC activity in this assay compared to a 0.6 μ/ml of a control antibody having a nonmutated IgG1 Fc region. Preferably the variant does not appear to have any CDC activity in the above CDC assay. [0085]
  • Preferably the polypeptide variant essentially retains the ability to bind antigen compared to the nonvariant polypeptide, i.e. the binding capability is no worse than about 20 fold, e.g. no worse than about 5 fold of that of the nonvariant polypeptide. The binding capability of the polypeptide variant may be determined using techniques such as fluorescence activated cell sorting (FACS) analysis or radioimmunoprecipitation (RIA), for example. [0086]
  • The ability of the polypeptide variant to bind an FcR may also be evaluated. Where the FcR is a high affinity Fc receptor, such as FcγRI or FcRn, binding can be measured by titrating monomeric polypeptide variant and measuring bound polypeptide variant using an antibody which specifically binds to the polypeptide variant in a standard ELISA format (see Example 2 below). Another FcR binding assay for low affinity FcRs is elaborated in more detail in the following section. [0087]
  • Preferably the variant retains the ability to bind one or more FcRs, e.g. the ability of the polypeptide variant to bind FcγRI, FcγRII, FcγRIII and/or FcRn is no more than about 20 fold reduced, preferably no more than about 10 fold reduced, and most preferably no more than about two fold reduced compared to the starting polypeptide as determined in the FcγRI or FcRn assays of Example 2 or the FcγRII or FcγRIII assays described in the following section. [0088]
  • A. Receptor Binding Assay and Immune Complex [0089]
  • A receptor binding assay has been developed herein which is particularly useful for determining binding of an analyte of interest to a receptor where the affinity of the analyte for the receptor is relatively weak, e.g. in the micromolar range as is the case for FcγRIIa, FcγRIIb, FcγRIIIa and FcγRIIIb. The method involves the formation of a molecular complex that has an improved avidity for the receptor of interest compared to the noncomplexed analyte. The preferred molecular complex is an immune complex comprising: (a) an Fc region-containing polypeptide (such as an antibody or an immunoadhesin); (b) a first target molecule which comprises at least two binding sites for the Fc region-containing polypeptide; and (c) a second target molecule which comprises at least two binding sites for the first target molecule. [0090]
  • In Example 1 below, the Fc region-containing polypeptide is an anti-IgE antibody, such as the E27 antibody (FIGS. [0091] 4A-4B). E27, when mixed with human IgE at an 1:1 molar ratio, forms a stable hexamer consisting of three E27 molecules and three IgE molecules. In Example 1 below, the “first target molecule” is a chimeric form of IgE in which the Fab portion of an anti-VEGF antibody is fused to the human IgE Fc portion and the “second target molecule” is the antigen to which the Fab binds (i.e. VEGF). Each molecule of IgE binds two molecules of VEGF. VEGF also binds two molecules of IgE per molecule of VEGF. When recombinant human VEGF was added at a 2:1 molar ratio to IgE:E27 hexamers, the hexamers were linked into larger molecular weight complexes via the IgE:VEGF interaction (FIG. 5). The Fc region of the anti-IgE antibody of the resultant immune complex binds to FcR with higher avidity than either uncomplexed anti-IgE or anti-IgE:lgE hexamers.
  • Other forms of molecular complexes for use in the receptor assay are contemplated. Examples comprising only an Fc region-containing polypeptide:first target molecule combination include an immunoadhesin:ligand combination such as VEGF receptor (KDR)-immunoadhesin: VEGF and a full-length bispecific antibody (bsAb):first target molecule. A further example of an Fc region-containing polypeptide:first target molecule:second target molecule combination include a nonblocking antibody:soluble receptor:ligand combination such as anti-Trk antibody:soluble Trk receptor:neurotrophin (Urfer et al. [0092] J. Biol Chem. 273(10):5829-5840 (1998)).
  • Aside from use in a receptor binding assay, the immune complexes described above have further uses including evaluation of Fc region-containing polypeptide function and immune complex clearance in vivo. Hence, the immune complex may be administered to a mammal (e.g. in a pre-clinical animal study) and evaluated for its half-life etc. [0093]
  • To determine receptor binding, a polypeptide comprising at least the binding domain of the receptor of interest (e.g. the extracellular domain of an α subunit of an FcR) may be coated on solid phase, such as an assay plate. The binding domain of the receptor alone or a receptor-fusion protein may be coated on the plate using standard procedures. Examples of receptor-fusion proteins include receptor-glutathione S-transferase (GST) fusion protein, receptor-chitin binding domain fusion protein, receptor-hexaHis tag fusion protein (coated on glutathione, chitin, and nickel coated plates, respectively). Alternatively, a capture molecule may be coated on the assay plate and used to bind the receptor-fusion protein via the non-receptor portion of the fusion protein. Examples include anti-hexaHis F(ab′)[0094] 2 coated on the assay plate used to capture receptor-hexaHis tail fusion or anti-GST antibody coated on the assay plate used to capture a receptor-GST fusion. In other embodiments, binding to cells expressing at least the binding domain of the receptor may be evaluated. The cells may be naturally occurring hematopoietic cells that express the FcR of interest or may be transformed with nucleic acid encoding the FcR or a binding domain thereof such that the binding domain is expressed at the surface of the cell to be tested.
  • The immune complex described hereinabove is added to the receptor-coated plates and incubated for a sufficient period of time such that the analyte binds to the receptor. Plates may then be washed to remove unbound complexes, and binding of the analyte may be detected according to known methods. For example, binding may be detected using a reagent (e.g. an antibody or fragment thereof) which binds specifically to the analyte, and which is optionally conjugated with a detectable label (detectable labels and methods for conjugating them to polypeptides are described below in the section entitled “Non-Therapeutic Uses for the Polypeptide Variant”). [0095]
  • As a matter of convenience, the reagents can be provided in an assay kit, i.e., a packaged combination of reagents, for combination with the analyte in assaying the ability of the analyte to bind to a receptor of interest. The components of the kit will generally be provided in predetermined ratios. The kit may provide the first target molecule and/or the second target molecule, optionally complexed together. The kit may further include assay plates coated with the receptor or a binding domain thereof (e.g. the extracellular domain of the αsubunit of an FcR). Usually, other reagents, such as an antibody that binds specifically to the analyte to be assayed, labeled directly or indirectly with an enzymatic label, will also be provided in the kit. Where the detectable label is an enzyme, the kit will include substrates and cofactors required by the enzyme (e.g. a substrate precursor which provides the detectable chromophore or fluorophore). In addition, other additives may be included such as stabilizers, buffers (e.g. assay and/or wash lysis buffer) and the like. The relative amounts of the various reagents may be varied widely to provide for concentrations in solution of the reagents that substantially optimize the sensitivity of the assay. Particularly, the reagents may be provided as dry powders, usually lyophilized, including excipients that on dissolution will provide a reagent solution having the appropriate concentration. The kit also suitably includes instructions for carrying out the assay. [0096]
  • B. Antibody Preparation [0097]
  • In the preferred embodiment of the invention, the Fc region-containing polypeptide which is modified according to the teachings herein is an antibody. Techniques for producing antibodies follow: [0098]
  • (i) Antigen Selection and Preparation [0099]
  • Where the polypeptide is an antibody, it is directed against an antigen of interest. Preferably, the antigen is a biologically important polypeptide and administration of the antibody to a mammal suffering from a disease or disorder can result in a therapeutic benefit in that mammal. However, antibodies directed against nonpolypeptide antigens (such as tumor-associated glycolipid antigens; see US Patent 5,091,178) are also contemplated. [0100]
  • Where the antigen is a polypeptide, it may be a transmembrane molecule (e.g. receptor) or ligand such as a growth factor. Exemplary antigens include molecules such as renin; a growth hormone, including human growth hormone and bovine growth hormone; growth hormone releasing factor; parathyroid hormone; thyroid stimulating hormone; lipoproteins; alpha-1-antitrypsin; insulin A-chain; insulin B-chain; proinsulin; follicle stimulating hormone; calcitonin; luteinizing hormone; glucagon; clofting factors such as factor VIIIC, factor IX, tissue factor, and von Willebrands factor; anti-clofting factors such as Protein C; atrial natriuretic factor; lung surfactant; a plasminogen activator, such as urokinase or human urine or tissue-type plasminogen activator (t-PA); bombesin; thrombin; hemopoietic growth factor; tumor necrosis factor-alpha and -beta; enkephalinase; RANTES (regulated on activation normally T-cell expressed and secreted); human macrophage inflammatory protein (MlP-1-alpha); a serum albumin such as human serum albumin; Muellerian-inhibiting substance; relaxin A-chain; relaxin B-chain; prorelaxin; mouse gonadotropin-associated peptide; a microbial protein, such as beta-lactamase; DNase; IgE; a cytotoxic T-lymphocyte associated antigen (CTLA), such as CTLA-4; inhibin; activin; vascular endothelial growth factor (VEGF); receptors for hormones or growth factors; protein A or D; rheumatoid factors; a neurotrophic factor such as bone-derived neurotrophic factor (BDNF), neurotrophin-3, -4, -5, or -6 (NT-3, NT-4, NT-5, or NT-6), or a nerve growth factor such as NGF-β; platelet-derived growth factor (PDGF); fibroblast growth factor such as aFGF and bFGF; epidermal growth factor (EGF); transforming growth factor (TGF) such as TGF-alpha and TGF-beta, including TGF-,β1, TGF-,β2, TGF-β3, TGF-β4, or TGF-β5; insulin-like growth factor-I and -II (IGF-I and IGF-II); des(1-3)-IGF-I (brain IGF-I), insulin-like growth factor binding proteins; CD proteins such as CD3, CD4, CD8, CD19 and CD20; erythropoietin; osteoinductive factors; immunotoxins; a bone morphogenetic protein (BMP); an interferon such as interferon-alpha, -beta, and -gamma; colony stimulating factors (CSFs), e.g., M-CSF, GM-CSF, and G-CSF; interieukins (ILs), e.g., IL-1 to IL-10; superoxide dismutase; T-cell receptors; surface membrane proteins; decay accelerating factor; viral antigen such as, for example, a portion of the AIDS envelope; transport proteins; homing receptors; addressins; regulatory proteins; integrins such as CD11a, CD11b, CD11c, CD18, an ICAM, VLA-4 and VCAM; a tumor associated antigen such as HER2, HER3 or HER4 receptor; and fragments of any of the above-listed polypeptides. [0101]
  • Preferred molecular targets for antibodies encompassed by the present invention include CD proteins such as CD3, CD4, CD8, CD19, CD20 and CD34; members of the ErbB receptor family such as the EGF receptor, HER2, HER3 or HER4 receptor; cell adhesion molecules such as LFA-1, Mac1, p150.95, VLA-4, ICAM-1, VCAM and αv/β3 integrin including either α or β subunits thereof (e.g. anti-CD11[0102] a, anti-CD18 or anti-CD11 b antibodies); growth factors such as VEGF; IgE; blood group antigens; flk2/flt3 receptor; obesity (OB) receptor; mpl receptor; CTLA-4; protein C etc.
  • Soluble antigens or fragments thereof, optionally conjugated to other molecules, can be used as immunogens for generating antibodies. For transmembrane molecules, such as receptors, fragments of these (e.g. the extracellular domain of a receptor) can be used as the immunogen. Alternatively, cells expressing the transmembrane molecule can be used as the immunogen. Such cells can be derived from a natural source (e.g. cancer cell lines) or may be cells which have been transformed by recombinant techniques to express the transmembrane molecule. Other antigens and forms thereof useful for preparing antibodies will be apparent to those in the art. [0103]
  • (ii) Polyclonal antibodies [0104]
  • Polyclonal antibodies are preferably raised in animals by multiple subcutaneous (sc) or intraperitoneal (ip) injections of the relevant antigen and an adjuvant. It may be useful to conjugate the relevant antigen to a protein that is immunogenic in the species to be immunized, e.g., keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, or soybean trypsin inhibitor using a bifunctional or derivatizing agent, for example, maleimidobenzoyl sulfosuccinimide ester (conjugation through cysteine residues), N-hydroxysuccinimide (through lysine residues), glutaraldehyde, succinic anhydride, SOCl[0105] 2, or R1N═C═NR, where R and R1 are different alkyl groups.
  • Animals are immunized against the antigen, immunogenic conjugates, or derivatives by combining, e.g., 100 μg or 5 μg of the protein or conjugate (for rabbits or mice, respectively) with 3 volumes of Freund's complete adjuvant and injecting the solution intradermally at multiple sites. One month later the animals are boosted with 1/5 to 1/10 the original amount of peptide or conjugate in Freund's complete adjuvant by subcutaneous injection at multiple sites. Seven to 14 days later the animals are bled and the serum is assayed for antibody titer. Animals are boosted until the titer plateaus. Preferably, the animal is boosted with the conjugate of the same antigen, but conjugated to a different protein and/or through a different cross-linking reagent. Conjugates also can be made in recombinant cell culture as protein fusions. Also, aggregating agents such as alum are suitably used to enhance the immune response. [0106]
  • (iii) Monoclonal antibodies [0107]
  • Monoclonal antibodies may be made using the hybridoma method first described by Kohler et al, Nature, 256:495 (1975), or may be made by recombinant DNA methods (U.S. Pat. No. 4,816,567). [0108]
  • In the hybridoma method, a mouse or other appropriate host animal, such as a hamster or macaque monkey, is immunized as hereinabove described to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the protein used for immunization. Alternatively, lymphocytes may be immunized in vitro. Lymphocytes then are fused with myeloma cells using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell (Goding, [0109] Monoclonal Antibodies: Principles and Practice, pp.59-103 (Academic Press, 1986)).
  • The hybridoma cells thus prepared are seeded and grown in a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, parental myeloma cells. For example, if the parental myeloma cells lack the enzyme hypoxanthine guanine phosphoribosyl transferase (HGPRT or HPRT), the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine (HAT medium), which substances prevent the growth of HGPRT-deficient cells. [0110]
  • Preferred myeloma cells are those that fuse efficiently, support stable high-level production of antibody by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium. Among these, preferred myeloma cell lines are murine myeloma lines, such as those derived from MOPC-21 and MPC-11 mouse tumors available from the Salk Institute Cell Distribution Center, San Diego, Calif. USA, and SP-2 or X63-Ag8-653 cells available from the American Type Culture Collection, Rockville, Md. USA. Human myeloma and mouse-human heteromyeloma cell lines also have been described for the production of human monoclonal antibodies (Kozbor, [0111] J. Immunol., 133:3001 (1984); Brodeur et al, Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987)).
  • Culture medium in which hybridoma cells are growing is assayed for production of monoclonal antibodies directed against the antigen. Preferably, the binding specificity of monoclonal antibodies produced by hybridoma cells is determined by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA). [0112]
  • After hybridoma cells are identified that produce antibodies of the desired specificity, affinity, and/or activity, the clones may be subcloned by limiting dilution procedures and grown by standard methods (Goding, [0113] Monoclonal Antibodies: Principles and Practice, pp.59-103 (Academic Press, 1986)). Suitable culture media for this purpose include, for example, D-MEM or RPMI-1640 medium. In addition, the hybridoma cells may be grown in vivo as ascites tumors in an animal.
  • The monoclonal antibodies secreted by the subclones are suitably separated from the culture medium, ascites fluid, or serum by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxylapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography. [0114]
  • DNA encoding the monoclonal antibodies is readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of the monoclonal antibodies). The hybridoma cells serve as a preferred source of such DNA. Once isolated, the DNA may be placed into expression vectors, which are then transfected into host cells such as [0115] E. coi cells, simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin protein, to obtain the synthesis of monoclonal antibodies in the recombinant host cells. Recombinant production of antibodies will be described in more detail below.
  • In a further embodiment, antibodies or antibody fragments can be isolated from antibody phage libraries generated using the techniques described in McCafferty et al, [0116] Nature, 348:552-554 1990). Clackson et al., Nature, 352:624-628 (1991) and Marks et al, J. Mol. Biol., 222:581-597 (1991) describe the isolation of murine and human antibodies, respectively, using phage libraries. Subsequent publications describe the production of high affinity (nM range) human antibodies by chain shuffling (Marks et al, Bio/Technology, 10:779-783 (1992)), as well as combinatorial infection and in vivo recombination as a strategy for constructing very large phage libraries (Waterhouse et al., Nuc. Acids. Res., 21:2265-2266 (1993)). Thus, these techniques are viable alternatives to traditional monoclonal antibody hybridoma techniques for isolation of monoclonal antibodies.
  • The DNA also may be modified, for example, by substituting the coding sequence for human heavy- and light-chain constant domains in place of the homologous murine sequences (U.S. Pat. No. 4,816,567; Morrison, et al., [0117] Proc. Natl Acad. Sci. USA, 81:6851 (1984)), or by covalently joining to the immunoglobulin coding sequence all or part of the coding sequence for a non-immunoglobulin polypeptide.
  • Typically such non-immunoglobulin polypeptides are substituted for the constant domains of an antibody, or they are substituted for the variable domains of one antigen-combining site of an antibody to create a chimeric bivalent antibody comprising one antigen-combining site having specificity for an antigen and another antigen-combining site having specificity for a different antigen. [0118]
  • (iv) Humanized and Human Antibodies [0119]
  • A humanized antibody has one or more amino acid residues introduced into it from a source which is non-human. These non-human amino acid residues are often referred to as “import” residues, which are typically taken from an “import” variable domain. Humanization can be essentially performed following the method of Winter and co-workers (Jones et al., Nature, 321:522-525 (1986); Riechmann et al, [0120] Nature, 332:323-327 (1988); Verhoeyen et al, Science, 239:1534-1536 (1988)), by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody. Accordingly, such “humanized” antibodies are chimeric antibodies (U.S. Pat. No. 4,816,567) wherein substantially less than an intact human variable domain has been substituted by the corresponding sequence from a non-human species. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR residues are substituted by residues from analogous sites in rodent antibodies.
  • The choice of human variable domains, both light and heavy, to be used in making the humanized antibodies is very important to reduce antigenicity. According to the so-called “best-fit” method, the sequence of the variable domain of a rodent antibody is screened against the entire library of known human variable-domain sequences. The human sequence which is closest to that of the rodent is then accepted as the human framework (FR) for the humanized antibody (Sims et al, [0121] J. Immunol., 151:2296 (1993); Chothia et al., J. Mol Biol, 196:901 (1987)). Another method uses a particular framework derived from the consensus sequence of all human antibodies of a particular subgroup of light or heavy chains. The same framework may be used for several different humanized antibodies (Carter et al., Proc. Natl. Acad. Sci. USA, 89:4285 (1992); Presta et al., J. Immnol., 151:2623 (1993)).
  • It is further important that antibodies be humanized with retention of high affinity for the antigen and other favorable biological properties. To achieve this goal, according to a preferred method, humanized antibodies are prepared by a process of analysis of the parental sequences and various conceptual humanized products using three-dimensional models of the parental and humanized sequences. Three-dimensional immunoglobulin models are commonly available and are familiar to those skilled in the art. Computer programs are available which illustrate and display probable three-dimensional conformational structures of selected candidate immunoglobulin sequences. Inspection of these displays permits analysis of the likely role of the residues in the functioning of the candidate immunoglobulin sequence, i.e., the analysis of residues that influence the ability of the candidate immunoglobulin to bind its antigen. In this way, FR residues can be selected and combined from the recipient and import sequences so that the desired antibody characteristic, such as increased affinity for the target antigen(s), is achieved. In general, the CDR residues are directly and most substantially involved in influencing antigen binding. [0122]
  • Alternatively, it is now possible to produce transgenic animals (e.g., mice) that are capable, upon immunization, of producing a full repertoire of human antibodies in the absence of endogenous immunoglobulin production. For example, it has been described that the homozygous deletion of the antibody heavy-chain joining region (J[0123] H) gene in chimeric and germ-line mutant mice results in complete inhibition of endogenous antibody production. Transfer of the human germ-line immunoglobulin gene array in such germ-line mutant mice will result in the production of human antibodies upon antigen challenge. See, e.g., Jakobovits et al., Proc. Natl. Acad. Sci. USA, 90:2551 (1993); Jakobovits et al., Nature, 362:255-258 (1993); Bruggermann et al, Year in Immuno., 7:33 (1993); and Duchosal et al. Nature 355:258 (1992). Human antibodies can also be derived from phage-display libraries (Hoogenboom et al., J. MoL Biol., 227:381 (1991); Marks et al., J. Mol. Biol, 222:581-597 (1991); Vaughan et al. Nature Biotech 14:309 (1996)).
  • (v) Multispecific Antibodies [0124]
  • Multispecific antibodies have binding specificities for at least two different antigens. While such molecules normally will only bind two antigens (i.e. bispecific antibodies, BsAbs), antibodies with additional specificities such as trispecific antibodies are encompassed by this expression when used herein. Examples of BsAbs include those with one arm directed against a tumor cell antigen and the other arm directed against a cytotoxic trigger molecule such as anti-FcγRl/anti-CD15, anti-p185[0125] HER2/FcγRIII (CD16), anti-CD3/anti-malignant B-cell (1D10), anti-CD3/anti-p185HER2, anti-CD3/anti-p97, anti-CD3/anti-renal cell carcinoma, anti-CD3/anti-OVCAR-3, anti-CD3/L-D1 (anti-colon carcinoma), anti-CD3/anti-melanocyte stimulating hormone analog, anti-EGF receptor/anti-CD3, anti-CD3/anti-CAMA1, anti-CD3/anti-CD19, anti-CD3/MoV18, anti-neural cell ahesion molecule (NCAM)/anti-CD3, anti-folate binding protein (FBP)/anti-CD3, anti-pan carcinoma associated antigen (AMOC-31)/anti-CD3; BsAbs with one arm which binds specifically to a tumor antigen and one arm which binds to a toxin such as anti-saporin/anti-ld-1, anti-CD22/anti-saporin, anti-CD7/anti-saporin, anti-CD38/anti-saporin, anti-CEA/anti-ricin A chain, anti-interferon-α(IFN-α)/anti-hybridoma idiotype, anti-CEA/anti-vinca alkaloid; BsAbs for converting enzyme activated prodrugs such as anti-CD30/anti-alkaline phosphatase (which catalyzes conversion of mitomycin phosphate prodrug to mitomycin alcohol); BsAbs which can be used as fibrinolytic agents such as anti-fibrin/anti-tissue plasminogen activator (tPA), anti-fibrin/anti-urokinase-type plasminogen activator (uPA); BsAbs for targeting immune complexes to cell surface receptors such as anti-low density lipoprotein (LDL)/anti-Fc receptor (e.g. FcγRI, FcγRII or FcγRIII); BsAbs for use in therapy of infectious diseases such as anti-CD3/anti-herpes simplex virus (HSV), anti-T-cell receptor:CD3 complex/anti-influenza, anti-FcγR/anti-HIV; BsAbs for tumor detection in vitro or in vivo such as anti-CEA/anti-EOTUBE, anti-CEA/anti-DPTA, anti-p185 HER2/anti-hapten; BsAbs as vaccine adjuvants; and BsAbs as diagnostic tools such as anti-rabbit IgG/anti-ferritin, anti-horse radish peroxidase (HRP)/anti-hormone, anti-somatostatin/anti-substance P, anti-HRP/anti-FITC, anti-CEA/anti-β-galactosidase. Examples of trispecific antibodies include anti-CD3/anti-CD4/anti-CD37, anti-CD3/anti-CD5/anti-CD37 and anti-CD3/anti-CD8/anti-CD37. Bispecific antibodies can be prepared as full length antibodies or antibody fragments (e.g. F(ab′)2 bispecific antibodies).
  • Methods for making bispecific antibodies are known in the art. Traditional production of full length bispecific antibodies is based on the coexpression of two immunoglobulin heavy chain-light chain pairs, where the two chains have different specificities (Millstein et al., [0126] Nature, 305:537-539 (1983)). Because of the random assortment of immunoglobulin heavy and light chains, these hybridomas (quadromas) produce a potential mixture of 10 different antibody molecules, of which only one has the correct bispecific structure. Purification of the correct molecule, which is usually done by affinity chromatography steps, is rather cumbersome, and the product yields are low. Similar procedures are disclosed in WO 93/08829, and in Traunecker et al., EMBO J., 10:3655-3659(1991).
  • According to a different approach, antibody variable domains with the desired binding specificities (antibody-antigen combining sites) are fused to immunoglobulin constant domain sequences. The fusion preferably is with an immunoglobulin heavy chain constant domain, comprising at least part of the hinge, CH2, and CH3 regions. It is preferred to have the first heavy-chain constant region (CH1) containing the site necessary for light chain binding, present in at least one of the fusions. DNAs encoding the immunoglobulin heavy chain fusions and, if desired, the immunoglobulin light chain, are inserted into separate expression vectors, and are co-transfected into a suitable host organism. This provides for great flexibility in adjusting the mutual proportions of the three polypeptide fragments in embodiments when unequal ratios of the three polypeptide chains used in the construction provide the optimum yields. It is, however, possible to insert the coding sequences for two or all three polypeptide chains in one expression vector when the expression of at least two polypeptide chains in equal ratios results in high yields or when the ratios are of no particular significance. [0127]
  • In a preferred embodiment of this approach, the bispecific antibodies are composed of a hybrid immunoglobulin heavy chain with a first binding specificity in one arm, and a hybrid immunoglobulin heavy chain-light chain pair (providing a second binding specificity) in the other arm. It was found that this asymmetric structure facilitates the separation of the desired bispecific compound from unwanted immunoglobulin chain combinations, as the presence of an immunoglobulin light chain in only one half of the bispecific molecule provides for a facile way of separation. This approach is disclosed in WO 94/04690. For further details of generating bispecific antibodies see, for example, Suresh et al., [0128] Methods in Enzymology, 121:210 (1986).
  • According to another approach described in W096/27011, the interface between a pair of antibody molecules can be engineered to maximize the percentage of heterodimers which are recovered from recombinant cell culture. The preferred interface comprises at least a part of the [0129] C H 3 domain of an antibody constant domain. In this method, one or more small amino acid side chains from the interface of the first antibody molecule are replaced with larger side chains (e.g. tyrosine or tryptophan). Compensatory “cavities” of identical or similar size to the large side chain(s) are created on the interface of the second antibody molecule by replacing large amino acid side chains with smaller ones (e.g. alanine or threonine). This provides a mechanism for increasing the yield of the heterodimer over other unwanted end-products such as homodimers.
  • Bispecific antibodies include cross-linked or “heteroconjugate” antibodies. For example, one of the antibodies in the heteroconjugate can be coupled to avidin, the other to biotin. Such antibodies have, for example, been proposed to target immune system cells to unwanted cells (U.S. Pat. No. 4,676,980), and for treatment of HIV infection (WO 91/00360, WO 921200373, and EP 03089). Heteroconjugate antibodies may be made using any convenient cross-linking methods. Suitable cross-linking agents are well known in the art, and are disclosed in U.S. Pat. No. 4,676,980, along with a number of cross-linking techniques. [0130]
  • Antibodies with more than two valencies are contemplated. For example, trispecific antibodies can be prepared. Tutt et al [0131] J. Immunol. 147:60 (1991).
  • While the polypeptide of interest herein is preferably an antibody, other Fc region-containing polypeptides which can be modified according to the methods described herein are contemplated. An example of such a molecule is an immunoadhesin. [0132]
  • C. Immunoadhesin Preparation [0133]
  • The simplest and most straightforward immunoadhesin design combines the binding domain(s) of the adhesin (e.g. the extracellular domain (ECD) of a receptor) with the Fc region of an immunoglobulin heavy chain. Ordinarily, when preparing the immunoadhesins of the present invention, nucleic acid encoding the binding domain of the adhesin will be fused C-terminally to nucleic acid encoding the N-terminus of an immunoglobulin constant domain sequence, however N-terminal fusions are also possible. [0134]
  • Typically, in such fusions the encoded chimeric polypeptide will retain at least functionally active hinge, [0135] C H2 and C H3 domains of the constant region of an immunoglobulin heavy chain. Fusions are also made to the C-terminus of the Fc portion of a constant domain, or immediately N-terminal to the C H1 of the heavy chain or the corresponding region of the light chain. The precise site at which the fusion is made is not critical; particular sites are well known and may be selected in order to optimize the biological activity, secretion, or binding characteristics of the immunoadhesin.
  • In a preferred embodiment, the adhesin sequence is fused to the N-terminus of the Fc region of immunoglobulin G[0136] 1 (IgG1). It is possible to fuse the entire heavy chain constant region to the adhesin sequence. However, more preferably, a sequence beginning in the hinge region just upstream of the papain cleavage site which defines IgG Fc chemically (i.e. residue 216, taking the first residue of heavy chain constant region to be 114), or analogous sites of other immunoglobulins is used in the fusion. In a particularly preferred embodiment, the adhesin amino acid sequence is fused to (a) the hinge region and C H 2 and C H 3 or (b) the C H1, hinge, C H2 and C H3 domains, of an IgG heavy chain.
  • For bispecific immunoadhesins, the immunoadhesins are assembled as multimers, and particularly as heterodimers or heterotetramers. Generally, these assembled immunoglobulins will have known unit structures. A basic four chain structural unit is the form in which IgG, lgD, and IgE exist. A four chain unit is repeated in the higher molecular weight immunoglobulins; IgM generally exists as a pentamer of four basic units held together by disulfide bonds. IgA globulin, and occasionally IgG globulin, may also exist in multimeric form in serum. In the case of multimer, each of the four units may be the same or different. [0137]
  • Various exemplary assembled immunoadhesins within the scope herein are schematically diagrammed below: [0138]
  • (a) AC[0139] L-ACL;
  • (b) AC[0140] H-(ACH, ACL-ACH, ACL-VHCH, or VLCL-ACH);
  • (c) AC[0141] L-ACH-(ACL-ACH, ACL-VHCH, VLCL-ACH, or VLCL-VH CH)
  • (d) AC[0142] L-VHCH-(ACH, or ACL-VHCH, or VLCL-ACH);
  • (e) V[0143] LCL-ACH-(ACL-VHCH, or VLCL-ACH); and
  • (f) (A-Y)[0144] n-(VLCL-VHCH)2,
  • wherein each A represents identical or different adhesin amino acid sequences; [0145]
  • V[0146] L is an immunoglobulin light chain variable domain;
  • V[0147] H is an immunoglobulin heavy chain variable domain;
  • C[0148] L is an immunoglobulin light chain constant domain;
  • C[0149] H is an immunoglobulin heavy chain constant domain;
  • n is an integer greater than 1; [0150]
  • Y designates the residue of a covalent cross-linking agent. [0151]
  • In the interests of brevity, the foregoing structures only show key features; they do not indicate joining (J) or other domains of the immunoglobulins, nor are disulfide bonds shown. However, where such domains are required for binding activity, they shall be constructed to be present in the ordinary locations which they occupy in the immunoglobulin molecules. [0152]
  • Alternatively, the adhesin sequences can be inserted between immunoglobulin heavy chain and light chain sequences, such that an immunoglobulin comprising a chimeric heavy chain is obtained. In this embodiment, the adhesin sequences are fused to the 3′ end of an immunoglobulin heavy chain in each arm of an immunoglobulin, either between the hinge and the [0153] C H2 domain, or between the C H2 and C H3 domains. Similar constructs have been reported by Hoogenboom, et al., Mol. Immunol. 28:1027-1037 (1991).
  • Although the presence of an immunoglobulin light chain is not required in the immunoadhesins of the present invention, an immunoglobulin light chain might be present either covalently associated to an adhesin-immunoglobulin heavy chain fusion polypeptide, or directly fused to the adhesin. In the former case, DNA encoding an immunoglobulin light chain is typically coexpressed with the DNA encoding the adhesin-immunoglobulin heavy chain fusion protein. Upon secretion, the hybrid heavy chain and the light chain will be covalently associated to provide an immunoglobulin-like structure comprising two disulfide-linked immunoglobulin heavy chain-light chain pairs. Methods suitable for the preparation of such structures are, for example, disclosed in U.S. Pat. No. 4,816,567, issued Mar. 28, 1989. [0154]
  • lmmunoadhesins are most conveniently constructed by fusing the cDNA sequence encoding the adhesin portion in-frame to an immunoglobulin cDNA sequence. However, fusion to genomic immunoglobulin fragments can also be used (see, e.g. Aruffo et al, [0155] Cell61:1303-1313 (1990); and Stamenkovic et al, Cell 66:1133-1144 (1991)). The latter type of fusion requires the presence of Ig regulatory sequences for expression. cDNAs encoding IgG heavy-chain constant regions can be isolated based on published sequences from cDNA libraries derived from spleen or peripheral blood lymphocytes, by hybridization or by polymerase chain reaction (PCR) techniques. The cDNAs encoding the “adhesin” and the immunoglobulin parts of the immunoadhesin are inserted in tandem into a plasmid vector that directs efficient expression in the chosen host cells.
  • D. Vectors, Host Cells and Recombinant Methods [0156]
  • The invention also provides isolated nucleic acid encoding a polypeptide variant as disclosed herein, vectors and host cells comprising the nucleic acid, and recombinant techniques for the production of the polypeptide variant. [0157]
  • For recombinant production of the polypeptide variant, the nucleic acid encoding it is isolated and inserted into a replicable vector for further cloning (amplification of the DNA) or for expression. DNA encoding the polypeptide variant is readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the polypeptide variant). Many vectors are available. The vector components generally include, but are not limited to, one or more of the following: a signal sequence, an origin of replication, one or more marker genes, an enhancer element, a promoter, and a transcription termination sequence. [0158]
  • (i) Signal Sequence Component [0159]
  • The polypeptide variant of this invention may be produced recombinantly not only directly, but also as a fusion polypeptide with a heterologous polypeptide, which is preferably a signal sequence or other polypeptide having a specific cleavage site at the N-terminus of the mature protein or polypeptide. The heterologous signal sequence selected preferably is one that is recognized and processed (ie., cleaved by a signal peptidase) by the host cell. For prokaryotic host cells that do not recognize and process the native polypeptide variant signal sequence, the signal sequence is substituted by a prokaryotic signal sequence selected, for example, from the group of the alkaline phosphatase, penicillinase, Ipp, or heat-stable enterotoxin II leaders. For yeast secretion the native signal sequence may be substituted by, e.g., the yeast invertase leader, α factor leader (including Saccharomyces and Kluyveromyces α-factor leaders), or acid phosphatase leader, the [0160] C. albicans glucoamylase leader, or the signal described in WO 90113646. In mammalian cell expression, mammalian signal sequences as well as viral secretory leaders, for example, the herpes simplex gD signal, are available.
  • The DNA for such precursor region is ligated in reading frame to DNA encoding the polypeptide variant. [0161]
  • (ii) Origin of Replication Component Both Expression and Cloning Vectors Contain a nucleic acid sequence that enables the vector to replicate in one or more selected host cells. Generally, in cloning vectors this sequence is one that enables the vector to replicate independently of the host chromosomal DNA, and includes origins of replication or autonomously replicating sequences. Such sequences are well known for a variety of bacteria, yeast, and viruses. The origin of replication from the plasmid pBR322 is suitable for most Gram-negative bacteria, the 2μ plasmid origin is suitable for yeast, and various viral origins (SV40, polyoma, adenovirus, VSV or BPV) are useful for cloning vectors in mammalian cells. Generally, the origin of replication component is not needed for mammalian expression vectors (the SV40 origin may typically be used only because it contains the early promoter). [0162]
  • (iii) Selection gene component [0163]
  • Expression and cloning vectors may contain a selection gene, also termed a selectable marker. Typical selection genes encode proteins that (a) confer resistance to antibiotics or other toxins, e.g., ampicillin, neomycin, methotrexate, or tetracycline, (b) complement auxotrophic deficiencies, or (c) supply critical nutrients not available from complex media, e.g., the gene encoding D-alanine racemase for Bacilli. [0164]
  • One example of a selection scheme utilizes a drug to arrest growth of a host cell. Those cells that are successfully transformed with a heterologous gene produce a protein conferring drug resistance and thus survive the selection regimen. Examples of such dominant selection use the drugs neomycin, mycophenolic acid and hygromycin. [0165]
  • Another example of suitable selectable markers for mammalian cells are those that enable the identification of cells competent to take up the polypeptide variant nucleic acid, such as DHFR, thymidine kinase, metallothionein-I and -II, preferably primate metallothionein genes, adenosine deaminase, ornithine decarboxylase, etc. [0166]
  • For example, cells transformed with the DHFR selection gene are first identified by culturing all of the transformants in a culture medium that contains methotrexate (Mtx), a competitive antagonist of DHFR. An appropriate host cell when wild-type DHFR is employed is the Chinese hamster ovary (CHO) cell line deficient in DHFR activity. [0167]
  • Alternatively, host cells (particularly wild-type hosts that contain endogenous DHFR) transformed or co-transformed with DNA sequences encoding polypeptide variant, wild-type DHFR protein, and another selectable marker such as [0168] aminoglycoside 3′-phosphotransferase (APH) can be selected by cell growth in medium containing a selection agent for the selectable marker such as an aminoglycosidic antibiotic, e.g., kanamycin, neomycin, or G418. See U.S. Pat. No. 4,965,199.
  • A suitable selection gene for use in yeast is the trpl gene present in the yeast plasmid YRp7 (Stinchcomb et al., [0169] Nature, 282:39 (1979)). The trpl gene provides a selection marker for a mutant strain of yeast lacking the ability to grow in tryptophan, for example, ATCC No. 44076 or PEP4-1. Jones, Genetics, 85:12 (1977). The presence of the trp1 lesion in the yeast host cell genome then provides an effective environment for detecting transformation by growth in the absence of tryptophan. Similarly, Leu2-deficient yeast strains (ATCC 20,622 or 38,626) are complemented by known plasmids bearing the Leu2 gene.
  • In addition, vectors derived from the 1.6 μm circular plasmid pKDl can be used for transformation of Kluyveromyces yeasts. Alternatively, an expression system for large-scale production of recombinant calf chymosin was reported for K. lactis. Van den Berg, [0170] Bio/Technology, 8:135 (1990). Stable multi-copy expression vectors for secretion of mature recombinant human serum albumin by industrial strains of Kluyveromyces have also been disclosed. Fleer et al., Bio/Technology, 9:968-975 (1991).
  • (iv) Promoter component [0171]
  • Expression and cloning vectors usually contain a promoter that is recognized by the host organism and is operably linked to the polypeptide variant nucleic acid. Promoters suitable for use with prokaryotic hosts include the phoA promoter, β-lactamase and lactose promoter systems, alkaline phosphatase, a tryptophan (trp) promoter system, and hybrid promoters such as the tac promoter. However, other known bacterial promoters are suitable. Promoters for use in bacterial systems also will contain a Shine-Dalgarno (S.D.) sequence operably linked to the DNA encoding the polypeptide variant. [0172]
  • Promoter sequences are known for eukaryotes. Virtually all eukaryotic genes have an AT-rich region located approximately 25 to 30 bases upstream from the site where transcription is initiated. Another sequence found 70 to 80 bases upstream from the start of transcription of many genes is a CNCAAT region where N may be any nucleotide. At the 3′ end of most eukaryotic genes is an AATAAA sequence that may be the signal for addition of the poly A tail to the 3′ end of the coding sequence. All of these sequences are suitably inserted into eukaryotic expression vectors. [0173]
  • Examples of suitable promoting sequences for use with yeast hosts include the promoters for 3-phosphoglycerate kinase or other glycolytic enzymes, such as enolase, glyceraldehyde-3-phosphate dehydrogenase, hexokinase, pyruvate decarboxylase, phosphofructokinase, glucose-6-phosphate isomerase, 3-phosphoglycerate mutase, pyruvate kinase, triosephosphate isomerase, phosphoglucose isomerase, and glucokinase. [0174]
  • Other yeast promoters, which are inducible promoters having the additional advantage of transcription controlled by growth conditions, are the promoter regions for [0175] alcohol dehydrogenase 2, isocytochrome C, acid phosphatase, degradative enzymes associated with nitrogen metabolism, metallothionein, glyceraldehyde-3-phosphate dehydrogenase, and enzymes responsible for maltose and galactose utilization. Suitable vectors and promoters for use in yeast expression are further described in EP 73,657. Yeast enhancers also are advantageously used with yeast promoters.
  • Polypeptide variant transcription from vectors in mammalian host cells is controlled, for example, by promoters obtained from the genomes of viruses such as polyoma virus, fowlpox virus, adenovirus (such as Adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-B virus and most preferably Simian Virus 40 (SV40), from heterologous mammalian promoters, e.g., the actin promoter or an immunoglobulin promoter, from heat-shock promoters, provided such promoters are compatible with the host cell systems. [0176]
  • The early and late promoters of the SV40 virus are conveniently obtained as an SV40 restriction fragment that also contains the SV40 viral origin of replication. The immediate early promoter of the human cytomegalovirus is conveniently obtained as a HindIII E restriction fragment. A system for expressing DNA in mammalian hosts using the bovine papilloma virus as a vector is disclosed in U.S. Pat. No. 4,419,446. A modification of this system is described in U.S. Pat. No. 4,601,978. See also Reyes et al, [0177] Nature 297:598-601 (1982) on expression of human β-interferon cDNA in mouse cells under the control of a thymidine kinase promoter from herpes simplex virus. Alternatively, the rous sarcoma virus long terminal repeat can be used as the promoter.
  • (v) Enhancer Element Component [0178]
  • Transcription of a DNA encoding the polypeptide variant of this invention by higher eukaryotes is often increased by inserting an enhancer sequence into the vector. Many enhancer sequences are now known from mammalian genes (globin, elastase, albumin, α-fetoprotein, and insulin). Typically, however, one will use an enhancer from a eukaryotic cell virus. Examples include the SV40 enhancer on the late side of the replication origin (bp 100-270), the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, and adenovirus enhancers. See also Yaniv, [0179] Nature 297:17-18 (1982) on enhancing elements for activation of eukaryotic promoters. The enhancer may be spliced into the vector at a position 5′ or 3′ to the polypeptide variant-encoding sequence, but is preferably located at a site 5′ from the promoter.
  • (vi) Transcription Termination Component [0180]
  • Expression vectors used in eukaryotic host cells (yeast, fungi, insect, plant, animal, human, or nucleated cells from other multicellular organisms) will also contain sequences necessary for the termination of transcription and for stabilizing the mRNA. Such sequences are commonly available from the 5′ and, occasionally 3′ , untranslated regions of eukaryotic or viral DNAs or cDNAs. These regions contain nucleotide segments transcribed as polyadenylated fragments in the untranslated portion of the mRNA encoding the polypeptide variant. One useful transcription termination component is the bovine growth hormone polyadenylation region. See WO94/11026 and the expression vector disclosed therein. [0181]
  • (vii) Selection and Transformation of Host Cells [0182]
  • Suitable host cells for cloning or expressing the DNA in the vectors herein are the prokaryote, yeast, or higher eukaryote cells described above. Suitable prokaryotes for this purpose include eubacteria, such as Gram-negative or Gram-positive organisms, for example, Enterobacteriaceae such as Escherichia, e.g., [0183] E coli, Enterobacter, Erwinia, Klebsiella, Proteus, Salmonella, e.g., Salmonella typhimurium, Serratia, e.g., Serratia marcescans, and Shigella, as well as Bacilli such as B. subtilis and B. licheniformis (e.g., B. licheniformis 41 P disclosed in DD 266,710 published Apr. 12, 1989), Pseudomonas such as P. aeruginosa, and Streptomyces. One preferred E. coil cloning host is E. coli 294 (ATCC 31,446), although other strains such as E. coli B, E. coil X1776 (ATCC 31,537), and E. coli W3110 (ATCC 27,325) are suitable. These examples are illustrative rather than limiting.
  • In addition to prokaryotes, eukaryotic microbes such as filamentous fungi or yeast are suitable cloning or expression hosts for polypeptide variant-encoding vectors. [0184] Saccharomyces cerevisiae, or common baker's yeast, is the most commonly used among lower eukaryotic host microorganisms. However, a number of other genera, species, and strains are commonly available and useful herein, such as Schizosaccharomyces pombe; Kluyveromyces hosts such as, e.g., K lactis, K. fragilis (ATCC 12,424), K. bulgaricus (ATCC 16,045), K. wickeramii (ATCC 24,178), K. waltii (ATCC 56,500), K. drosophilarum (ATCC 36,906), K. thermotolerans, and K. marxianus; yarrowia (EP 402,226); Pichia pastoris (EP 183,070); Candida; Trichoderma reesia (EP 244,234); Neurospora crassa; Schwanniomyces such as Schwanniomyces occidentalis; and filamentous fungi such as, e.g., Neurospora, Penicillium, Tolypocladium, and Aspergillus hosts such as A. nidulans and A. niger.
  • Suitable host cells for the expression of glycosylated polypeptide variant are derived from multicellular organisms. Examples of invertebrate cells include plant and insect cells. Numerous baculoviral strains and variants and corresponding permissive insect host cells from hosts such as [0185] Spodoptera frugiperda (caterpillar), Aedes aegypti (mosquito), Aedes albopictus (mosquito), Drosophila melanogaster (fruiffly), and Bombyx mori have been identified. A variety of viral strains for transfection are publicly available, e.g., the L-1 variant of Autographa californica NPV and the Bm-5 strain of Bombyx mori NPV, and such viruses may be used as the virus herein according to the present invention, particularly for transfection of Spodoptera frugiperda cells.
  • Plant cell cultures of cotton, corn, potato, soybean, petunia, tomato, and tobacco can also be utilized as hosts. [0186]
  • However, interest has been greatest in vertebrate cells, and propagation of vertebrate cells in culture (tissue culture) has become a routine procedure. Examples of useful mammalian host cell lines are monkey kidney CV1 line transformed by SV40 (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture, Graham et al., [0187] J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK, ATCC CCL 10); Chinese hamster ovary cells/-DHFR (CHO, Urlaub et al, Proc. Natl. Acad. Sci. USA 77:4216 (1980)); mouse sertoli cells (TM4, Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CV1 ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL-1587); human cervical carcinoma cells (HELA, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); buffalo rat liver cells (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells (Hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL51); TRI cells (Mather et al., Annals N.Y. Acad. Sci. 383:44-68 (1982)); MRC 5 cells; FS4 cells; and a human hepatoma line (Hep G2).
  • Host cells are transformed with the above-described expression or cloning vectors for polypeptide variant production and cultured in conventional nutrient media modified as appropriate for inducing promoters, selecting transformants, or amplifying the genes encoding the desired sequences. [0188]
  • (viii) Culturing the Host Cells [0189]
  • The host cells used to produce the polypeptide variant of this invention may be cultured in a variety of media. Commercially available media such as Ham's F10 (Sigma), Minimal Essential Medium ((MEM), (Sigma), RPMI-1640 (Sigma), and Dulbecco's Modified Eagle's Medium ((DMEM), Sigma) are suitable for culturing the host cells. In addition, any of the media described in Ham et al, [0190] Meth. Enz. 58:44 (1979), Barnes et al., Anal Biochem. 102:255 (1980), U.S. Pat. Nos. 4,767,704; 4,657,866; 4,927,762; 4,560,655; or 5,122,469; WO 90/03430; WO 87/00195; or U.S. Pat. No. Re. 30,985 may be used as culture media for the host cells. Any of these media may be supplemented as necessary with hormones and/or other growth factors (such as insulin, transferrin, or epidermal growth factor), salts (such as sodium chloride, calcium, magnesium, and phosphate), buffers (such as HEPES), nucleotides (such as adenosine and thymidine), antibiotics (such as GENTAMYCIN™ drug), trace elements (defined as inorganic compounds usually present at final concentrations in the micromolar range), and glucose or an equivalent energy source. Any other necessary supplements may also be included at appropriate concentrations that would be known to those skilled in the art. The culture conditions, such as temperature, pH, and the like, are those previously used with the host cell selected for expression, and will be apparent to the ordinarily skilled artisan.
  • (ix) Polypeptide Variant Purification [0191]
  • When using recombinant techniques, the polypeptide variant can be produced intracellularly, in the periplasmic space, or directly secreted into the medium. If the polypeptide variant is produced intracellularly, as a first step, the particulate debris, either host cells or lysed fragments, is removed, for example, by centrifugation or ultrafiltration. Carter et al., [0192] Bio/lTechnology 10:163-167 (1992) describe a procedure for isolating antibodies which are secreted to the periplasmic space of E coil. Briefly, cell paste is thawed in the presence of sodium acetate (pH 3.5), EDTA, and phenylmethylsulfonylfluoride (PMSF) over about 30 min. Cell debris can be removed by centrifugation. Where the polypeptide variant is secreted into the medium, supernatants from such expression systems are generally first concentrated using a commercially available protein concentration filter, for example, an Amicon or Millipore Pellicon ultrafiltration unit. A protease inhibitor such as PMSF may be included in any of the foregoing steps to inhibit proteolysis and antibiotics may be included to prevent the growth of adventitious contaminants.
  • The polypeptide variant composition prepared from the cells can be purified using, for example, hydroxylapatite chromatography, gel electrophoresis, dialysis, and affinity chromatography, with affinity chromatography being the preferred purification technique. The suitability of protein A as an affinity ligand depends on the species and isotype of any immunoglobulin Fc region that is present in the polypeptide variant. Protein A can be used to purify polypeptide variants that are based on human γ1, γ2, or γ4 heavy chains (Lindmark et al, [0193] J. Immunol. Meth. 62:1-13 (1983)).
  • Protein G is recommended for all mouse isotypes and for human γ3 (Guss et al, [0194] EMBO J. 5:15671575 (1986)). The matrix to which the affinity ligand is attached is most often agarose, but other matrices are available. Mechanically stable matrices such as controlled pore glass or poly(styrenedivinyl)benzene allow for faster flow rates and shorter processing times than can be achieved with agarose. Where the polypeptide variant comprises a C H3 domain, the Bakerbond ABX™ resin (J. T. Baker, Phillipsburg, N.J.) is useful for purification. Other techniques for protein purification such as fractionation on an ion-exchange column, ethanol precipitation, Reverse Phase HPLC, chromatography on silica, chromatography on heparin SEPHAROSE™ chromatography on an anion or cation exchange resin (such as a polyaspartic acid column), chromatofocusing, SDS-PAGE, and ammonium sulfate precipitation are also available depending on the polypeptide variant to be recovered.
  • Following any preliminary purification step(s), the mixture comprising the polypeptide variant of interest and contaminants may be subjected to low pH hydrophobic interaction chromatography using an elution buffer at a pH between about 2.5-4.5, preferably performed at low salt concentrations (e.g.,from about 0-0.25M salt). [0195]
  • E. Pharmaceutical Formulations [0196]
  • Therapeutic formulations of the polypeptide variant are prepared for storage by mixing the polypeptide variant having the desired degree of purity with optional physiologically acceptable carriers, excipients or stabilizers ([0197] Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)), in the form of lyophilized formulations or aqueous solutions. Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyidimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptide; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g., Zn-protein complexes); and/or non-ionic surfactants such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG).
  • The formulation herein may also contain more than one active compound as necessary for the particular indication being treated, preferably those with complementary activities that do not adversely affect each other. Such molecules are suitably present in combination in amounts that are effective for the purpose intended. [0198]
  • The active ingredients may also be entrapped in microcapsule prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsule and poly-(methylmethacylate) microcapsule, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules) or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980). [0199]
  • The formulations to be used for in vivo administration must be sterile. This is readily accomplished by filtration through sterile filtration membranes. [0200]
  • Sustained-release preparations may be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the polypeptide variant, which matrices are in the form of shaped articles, e.g., films, or microcapsule. Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate), or poly(vinylalcohol)), polylactides (U.S. Pat. No. 3,773,919), copolymers of L-glutamic acid and γ ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the LUPRON DEPOT™ (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-(−)-3-hydroxybutyric acid. While polymers such as ethylene-vinyl acetate and lactic acid-glycolic acid enable release of molecules for over 100 days, certain hydrogels release proteins for shorter time periods. When encapsulated antibodies remain in the body for a long time, they may denature or aggregate as a result of exposure to moisture at 37°C., resulting in a loss of biological activity and possible changes in immunogenicity. Rational strategies can be devised for stabilization depending on the mechanism involved. For example, if the aggregation mechanism is discovered to be intermolecular S-S bond formation through thio-disulfide interchange, stabilization may be achieved by modifying sulihydryl residues, lyophilizing from acidic solutions, controlling moisture content, using appropriate additives, and developing specific polymer matrix compositions. [0201]
  • F. Non-Therapeutic Uses for the Polypeptide Variant [0202]
  • The polypeptide variant of the invention may be used as an affinity purification agent. In this process, the polypeptide variant is immobilized on a solid phase such a Sephadex resin or filter paper, using methods well known in the art. The immobilized polypeptide variant is contacted with a sample containing the antigen to be purified, and thereafter the support is washed with a suitable solvent that will remove substantially all the material in the sample except the antigen to be purified, which is bound to the immobilized polypeptide variant. Finally, the support is washed with another suitable solvent, such as glycine buffer, pH 5.0, that will release the antigen from the polypeptide variant. [0203]
  • The polypeptide variant may also be useful in diagnostic assays, e.g., for detecting expression of an antigen of interest in specific cells, tissues, or serum. [0204]
  • For diagnostic applications, the polypeptide variant typically will be labeled with a detectable moiety. Numerous labels are available which can be generally grouped into the following categories: [0205]
  • (a) Radioisotopes, such as [0206] 35S, 14C, 125I, 3H, and 131I. The polypeptide variant can be labeled with the radioisotope using the techniques described in Current Protocots in Immunology, Volumes 1 and 2, Coligen et al., Ed. Wiley-lnterscience, New York, New York, Pubs. (1991) for example and radioactivity can be measured using scintillation counting.
  • (b) Fluorescent labels such as rare earth chelates (europium chelates) or fluorescein and its derivatives, rhodamine and its derivatives, dansyl, Lissamine, phycoerythrin and Texas Red are available. The fluorescent labels can be conjugated to the polypeptide variant using the techniques disclosed in Current Protocols in Immunology, supra, for example. Fluorescence can be quantified using a fluorimeter. [0207]
  • (c) Various enzyme-substrate labels are available and U.S. Pat. No. 4,275,149 provides a review of some of these. The enzyme generally catalyzes a chemical alteration of the chromogenic substrate that can be measured using various techniques. For example, the enzyme may catalyze a color change in a substrate, which can be measured spectrophotometrically. Alternatively, the enzyme may alter the fluorescence or chemiluminescence of the substrate. Techniques for quantifying a change in fluorescence are described above. The chemiluminescent substrate becomes electronically excited by a chemical reaction and may then emit light which can be measured (using a chemiluminometer, for example) or donates energy to a fluorescent acceptor. Examples of enzymatic labels include luciferases (e.g., firefly luciferase and bacterial luciferase; U.S. Pat. No. 4,737,456), luciferin, 2,3-dihydrophthalazinediones, malate dehydrogenase, urease, peroxidase such as horseradish peroxidase (HRPO), alkaline phosphatase, β-galactosidase, glucoamylase, lysozyme, saccharide oxidases (e.g., glucose oxidase, galactose oxidase, and glucose-6-phosphate dehydrogenase), heterocyclic oxidases (such as uricase and xanthine oxidase), lactoperoxidase, microperoxidase, and the like. Techniques for conjugating enzymes to antibodies are described in O′Sullivan et al., Methods for the Preparation of Enzyme-Antibody Conjugates for use in Enzyme Immunoassay, in Methods in Enzym. (ed J. Langone & H. Van Vunakis), Academic press, New York, 73:147-166 (1981). [0208]
  • Examples of enzyme-substrate combinations include, for example: [0209]
  • (i) Horseradish peroxidase (HRPO) with hydrogen peroxidase as a substrate, wherein the hydrogen peroxidase oxidizes a dye precursor (e.g.,orthophenylene diamine (OPD) or 3,3′,5,5′-tetramethyl benzidine hydrochloride (TMB)); [0210]
  • (ii) alkaline phosphatase (AP) with para-Nitrophenyl phosphate as chromogenic substrate; and [0211]
  • (iii) β-D-galactosidase (β-D-Gal) with a chromogenic substrate (e.g., p-nitrophenyl-β-D-galactosidase) or fluorogenic substrate 4-methylumbelliferyl-β-D-galactosidase. [0212]
  • Numerous other enzyme-substrate combinations are available to those skilled in the art. For a general review of these, see U.S. Pat. Nos. 4,275,149 and 4,318,980. [0213]
  • Sometimes, the label is indirectly conjugated with the polypeptide variant. The skilled artisan will be aware of various techniques for achieving this. For example, the polypeptide variant can be conjugated with biotin and any of the three broad categories of labels mentioned above can be conjugated with avidin, or vice versa. Biotin binds selectively to avidin and thus, the label can be conjugated with the polypeptide variant in this indirect manner. Alternatively, to achieve indirect conjugation of the label with the polypeptide variant, the polypeptide variant is conjugated with a small hapten (e.g., digoxin) and one of the different types of labels mentioned above is conjugated with an anti-hapten polypeptide variant (e.g., anti-digoxin antibody). Thus, indirect conjugation of the label with the polypeptide variant can be achieved. [0214]
  • In another embodiment of the invention, the polypeptide variant need not be labeled, and the presence thereof can be detected using a labeled antibody which binds to the polypeptide variant. [0215]
  • The polypeptide variant of the present invention may be employed in any known assay method, such as competitive binding assays, direct and indirect sandwich assays, and immunoprecipitation assays. Zola, [0216] Monoclonal Antibodies:A Manualof Techniques, pp.147-158 (CRC Press, Inc. 1987).
  • The polypeptide variant may also be used for in vivo diagnostic assays. Generally, the polypeptide variant is labeled with a radionuclide (such as [0217] 111In, 99Tc, 14C, 131I, 125I, 3H, 32 P or 35S) so that the antigen or cells expressing it can be localized using immunoscintiography.
  • G. In Vivo Uses for the Polypeptide Variant [0218]
  • It is contemplated that the polypeptide variant of the present invention may be used to treat a mammal e.g. a patient suffering from a disease or disorder who could benefit from administration of the polypeptide variant. The conditions which can be treated with the polypeptide variant are many and include cancer (e.g. where the polypeptide variant binds the HER2 receptor or CD20); allergic conditions such as asthma (with an anti-IgE antibody); and LFA-mediated disorders (e.g. where the polypeptide variant is an anti-LFA-1 or anti-ICAM-1 antibody) etc. Where the polypeptide variant does not bind complement, but retains FcR binding capability, exemplary diseases or disorders to be treated include: cancer (e.g. where ADCC function is desirable, but complement activation would lead to amplified side effects, such as vasculitis in the blood vessels at the tumor site); disorders treated with an agonist antibody; disorders wherein the polypeptide variant binds a soluble antigen and wherein stoichiometry leads to immune complexes which activate the complement cascade and result in unwanted side effects; conditions employing an antagonist antibody which downmodulates receptor function without damaging tissue or organ function; [0219]
  • intravenous immunoglobulin treatment for, e.g., immunodeficient individuals with autoimmune disorders. [0220]
  • The polypeptide variant is administered by any suitable means, including parenteral, subcutaneous, intraperitoneal, intrapulmonary, and intranasal, and, if desired for local immunosuppressive treatment, intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. In addition, the polypeptide variant is suitably administered by pulse infusion, particularly with declining doses of the polypeptide variant. Preferably the dosing is given by injections, most preferably intravenous or subcutaneous injections, depending in part on whether the administration is brief or chronic. [0221]
  • For the prevention or treatment of disease, the appropriate dosage of polypeptide variant will depend on the type of disease to be treated, the severity and course of the disease, whether the polypeptide variant is administered for preventive or therapeutic purposes, previous therapy, the patient's clinical history and response to the polypeptide variant, and the discretion of the attending physician. The polypeptide variant is suitably administered to the patient at one time or over a series of treatments. [0222]
  • Depending on the type and severity of the disease, about 1 μg/kg to 15 mg/kg (e.g., 0.1-20 mg/kg) of polypeptide variant is an initial candidate dosage for administration to the patient, whether, for example, by one or more separate administrations, or by continuous infusion. A typical daily dosage might range from about 1 μg/kg to 100 mg/kg or more, depending on the factors mentioned above. For repeated administrations over several days or longer, depending on the condition, the treatment is sustained until a desired suppression of disease symptoms occurs. However, other dosage regimens may be useful. The progress of this therapy is easily monitored by conventional techniques and assays. [0223]
  • The polypeptide variant composition will be formulated, dosed, and administered in a fashion consistent with good medical practice. Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners. The “therapeutically effective amount” of the polypeptide variant to be administered will be governed by such considerations, and is the minimum amount necessary to prevent, ameliorate, or treat a disease or disorder. The polypeptide variant need not be, but is optionally formulated with one or more agents currently used to prevent or treat the disorder in question. The effective amount of such other agents depends on the amount of polypeptide variant present in the formulation, the type of disorder or treatment, and other factors discussed above. These are generally used in the same dosages and with administration routes as used hereinbefore or about from 1 to 99% of the heretofore employed dosages. [0224]
  • The invention will be more fully understood by reference to the following examples. They should not, however, be construed as limiting the scope of this invention. All literature and patent citations mentioned herein are expressly incorporated by reference. [0225]
    • EXAMPLE 1 Low Affinity Receptor Binding Assay
  • This assay determines binding of an IgG Fc region to recombinant FcγRIIa, FcγRIIb and FcγRIIIa α subunits expressed as His6-glutathione S transferase (GST)-tagged fusion proteins. Since the affinity of the Fc region of IgG1 for the FcγRI is in the nanomolar range, the binding of IgG1 Fc mutants can be measured by titrating monomeric IgG and measuring bound IgG with a polyclonal anti-lgG in a standard ELISA format (Example 2 below). The affinity of the other members of the FcγR family, ie. FcγRIIa, FcγRIIb and FcγRIIIa for IgG is however in the micromolar range and binding of monomeric IgG1 for these receptors can not be reliably measured in an ELISA format. [0226]
  • The following assay utilizes Fc mutants of recombinant anti-IgE E27 (FIGS. 4A and 4B) which, when mixed with human IgE at a 1:1 molar ratio, forms a stable hexamer consisting of three anti-IgE molecules and three IgE molecules. A recombinant chimeric form of IgE (chimeric IgE) was engineered and consists of a human IgE Fc region and the Fab of an anti-VEGF antibody (Presta et al [0227] Cancer Research 57:4593-4599 (1997)) which binds two VEGF molecules per mole of anti-VEGF. When recombinant human VEGF is added at a 2:1 molar ratio to chimeric IgE:E27 hexamers, the hexamers are linked into larger molecular weight complexes via the chimeric IgE Fab:VEGF interaction. The E27 component of this complex binds to the FcγRIIa, FcγRIIb and FcγIIIa α subunits with higher avidity to permit detection in an ELISA format.
    • MATERIALS AND METHODS
  • Receptor Coat. Fcγ receptor α subunits were expressed as GST fusions of His6 tagged extracellular domains (ECDs) in 293 cells resulting in an ECD-6His-GST fusion protein (Graham et al [0228] J. Gen. Virol. 36:59-72 (1977) and Gorman et al DNA Prot. Eng. Tech. 2:3-10 (1990)) and purified by Ni-NTA column chromatography (Qiagen, Australia) and buffer exchanged into phosphate buffered saline (PBS). Concentrations were determined by absorption at 280nm using extinction coefficients derived by amino acid composition analysis. Receptors were coated onto Nunc F96 maxisorb plates (cat no. 439454) at 100 ng per well by adding 100 μl of receptor-GST fusion at 1 μg/ml in PBS and incubated for 48 hours at 4° C. Prior to assay, plates are washed 3×0 with 250 μl of wash buffer (PBS pH 7.4 containing 0.5% TWEEN 20™) and blocked with 250 μl of assay buffer (50 mM Tris buffered saline, 0.05% TWEEN 20™, 0.5% RIA grade bovine albumin (Sigma A7888), and 2 mM EDTA pH 7.4).
  • Immune Complex Formation: Equal molar amounts (1:1) of E27 and recombinant chimeric IgE which binds two moles recombinant human VEGF per mole of chimeric IgE are added to a 12 ×75 mm polypropylene tube in PBS and mixed by rotation for 30 minutes at 25° C. E27 (anti-IgE) /chimeric IgE (IgE) hexamers are formed during this incubation. Recombinant human VEGF (165 form, MW 44,000) is added at a 2:1 molar ratio to the IgE concentration and mixed by rotation an additional 30 minutes at 25° C. VEGF- chimeric IgE binding links E27:chimeric IgE hexamers into larger molecular weight complexes which bind FcγR α subunit ECD coated plates via the Fc region of the E27 antibody. [0229]
  • E27:chimeric IgE:VEGF(1:1:2 molar ratio) complexes are added to FcγR α subunit coated plates at E27 concentrations of 5 μg and 1 μg total IgG in quadruplicate in assay buffer and incubated for 120 minutes at 25° C. on an orbital shaker. [0230]
  • Complex Detection: Plates are washed 5× with wash buffer to remove unbound complexes and IgG binding is detected by adding 100 μl horse radish peroxidase (HRP) conjugated goat anti-human IgG (γ) heavy chain specific (Boehringer Mannheim 1814249) at 1:10,000 in assay buffer and incubated for 90 min at 25° C. on an orbital shaker. Plates are washed 5× with wash buffer to remove unbound HRP goat anti-human IgG and bound anti-lgG is detected by adding 100 μl of substrate solution (0.4mg/ml o-phenylenedaimine dihydrochloride, Sigma P6912, 6 mM H[0231] 2O2 in PBS) and incubating for 8 min at 25° C. Enzymatic reaction is stopped by the addition of 100 μl 4.5N H2SO4 and colorimetric product is measured at 490 nm on a 96 well plate densitometer (Molecular Devices). Binding of E27 mutant complexes is expressed as a percent of the wild type E27 containing complex.
    • EXAMPLE 2 Identification of Unique C1q Binding Sites in a Human IgG Antibody
  • In the present study, mutations were identified in the CH2 domain of a human IgG1 antibody, “C2B8” (Reff et al., Blood 83:435 (1994)), that ablated binding of the antibody to C1q but did not alter the conformation of the antibody nor affect binding to each of the FcγRs. By alanine scanning mutagenesis, three mutants in human IgG1 were identified, K322A P329A, and P331, that were non-lytic and had decreased binding to C1q. The data suggested that the core C1q binding sites in human IgG1 is different from that of murine IgG2b. In addition, K322A, P329A and P331A were found to bind normally to the CD20 antigen, and to four Fc receptors, FcγRI, FcγRII, FcγRIII and FcRn. [0232]
    • MATERIALS AND METHODS
  • Construction of C2B8 Mutants: The chimeric light and heavy chains of anti-CD20 antibody C2B8 (Reff et al., [0233] Blood 83:435 (1994)) subcloned separately into previously described PRK vectors (Gorman et al, DNA Protein Eng. Tech. 2:3 (1990)) were used. By site directed mutagenesis (Kunkel et al., Proc. Natl. Acad.Sci.USA 82:488 (1985)), alanine scan variants of the Fc region in the heavy chain were constructed. The heavy and light chain plasmids were co-transfected into an adenovirus transformed human embryonic kidney cell line as previously described (Werther et al., J. Immunol 157:4986 (1996)). The media was changed to serum-free 24 hours after transfection and the secreted antibody was harvested after five days. The antibodies were purified using Protein A-SEPHAROSE CL-4B™ (Pharmacia), buffer exchanged and concentrated to 0.5 ml with PBS using a Centricon-30 (Amicon), and stored at 4° C. The concentration of the antibody was determined using total Ig-binding ELISA.
  • C1q Binding ELISA: Costar 96 well plates were coated overnight at 4° C. with the indicated concentrations of 2B8 in coating buffer (0.05 M sodium carbonate buffer), pH 9. The plates were then washed 3× with PBS/ 0.05[0234] % TWEEN 20™, pH 7.4 and blocked with 200 μl of ELISA diluent without thimerosal (0.1 M NaPO4/0.1M NaCl/0.1% gelatin/0.05% TWEEN 20 ™/ 0.05% ProClin300) for 1 hr at room temperature. The plate was washed 3× with wash buffer, an aliquot of 100 μl of 2 μg/ml C1q (Quidel, San Diego, Calif.) was added to each well and incubated for 2 hrs at room temperature. The plate was then washed 6× with wash buffer. 100 μl of a 1:1000 dilution of sheep anti-complement C1q peroxidase conjugated antibody (Biodesign) was added to each well and incubated for 1 hour at room temperature. The plate was again washed 6× with wash buffer and 100 μl of substrate buffer (PBS/0.012% H2O2) containing OPD (O-phenylenediamine dihydrochloride (Sigma)) was added to each well. The oxidation reaction, observed by the appearance of a yellow color, was allowed to proceed for 30 minutes and stopped by the addition of 100 μl of 4.5 N H2SO4. The absorbance was then read at (492-405) nm using a microplate reader (SPECTRA MAX 250™, Molecular Devices Corp.). The appropriate controls were run in parallel (i.e. the ELISA was performed without C1q for each concentration of C2B8 used and also the ELISA was performed without C2B8). For each mutant, C1q binding was measured by plotting the absorbance (492-405) nm versus concentration of C2B8 in μg/ml using a 4-parameter curve fitting program (KALEIDAGRAPHTM) and comparing EC50 values.
  • Complement Dependent Cytotoxicity (CDC) Assay: This assay was performed essentially as previously described (Gazzano-Santoro et al, [0235] J. Immunol. Methods 202:163 (1997)). Various concentrations of C2B8 (0.08-20 μg/ml) were diluted with RHB buffer (RPMI 1640/20 mM HEPES (pH 7.2)/2mM Glutamine/0.1% BSA/100 μml Gentamicin). Human complement (Quidel) was diluted 1:3 in RHB buffer and WIL2-S cells (available from the ATCC, Manassas, Va.) which express the CD20 antigen were diluted to a density of 1×106 cells /ml with RHB buffer. Mixtures of 150 μl containing equal volumes of C2B8, diluted human complement and WIL2-S cells were added to a flat bottom tissue culture 96 well plate and allowed to incubate for 2 hrs at 37° C. and 5% CO2 to facilitate complement mediated cell lysis. 50 μl of alamar blue (Accumed International) was then added to each well and incubated overnight at 37° C. The absorbance was measured using a 96-well fluorometer with excitation at 530 nm and emission at 590 nm. As described by Gazzano-Santoro et al., the results are expressed in relative fluorescence units (RFU). The sample concentrations were computed from a C2B8 standard curve and the percent activity as compared to wild type C2B8 is reported for each mutant.
  • CD20 Binding Potency of the C2B8 Mutants: The binding of C2B8 and mutants to the CD20 antigen were assessed by a method previously described (Reff et al, (1994), supra; reviewed in Gazzano-Santoro et al, (1996), supra). WIL2-S cells were grown for 3-4 days to a cell density of 1×10[0236] 6 cells/ml. The cells were washed and spun twice in FACS buffer (PBS/0.1% BSA/0.02% NaN3) and resuspended to a cell density of 5×106 cells/ml. 200 μl of cells (5×106 cells/ml) and 20 μl of diluted C2B8 samples were added to a 5 ml tube and incubated at room temperature for 30 minutes with agitation. The mixture was then washed with 2 ml of cold FACS buffer, spun down and resuspended in 200 μl of cold FACS buffer. To the suspension, 10 μl of goat anti-human IgG-FITC (American Qualex Labs.) was added and the mixture was incubated in the dark at room temperature for 30 minutes with agitation. After incubation, the mixture was washed with 2 ml of FACS buffer, spun down and resuspended in 1 ml of cold fixative buffer (1% formaldehyde in PBS). The samples were analyzed by flow cytometry and the results expressed as relative fluorescence units (RFU) were plotted against antibody concentrations using a 4-parameter curve fitting program (KALEIDAGRAPH™). The EC50 values are reported as a percentage of that of the C2B8 reference material.
  • FcγR Binding Eilsas: [0237]
  • FcγRI α subunit-GST fusion was coated onto Nunc F96 maxisorb plates (cat no. 439454) by adding 100 μl of receptor-GST fusion at 1 μg/ml in PBS and incubated for 48 hours at 4° C. Prior to assay, plates are washed 3× with 250 μl of wash buffer (PBS pH 7.4 containing 0.5[0238] % TWEEN 20™) and blocked with 250 μl of assay buffer (50 mM Tris buffered saline, 0.05% TWEEN 20™, 0.5% RIA grade bovine albumin (Sigma A7888), and 2 mM EDTA pH 7.4). Samples diluted to 10 μg/ml in 1 ml of assay buffer are added to FcγRI α subunit coated plates and incubated for 120 minutes at 25° C. on an orbital shaker. Plates are washed 5× with wash buffer to remove unbound complexes and IgG binding is detected by adding 100 μl horse radish peroxidase (HRP) conjugated goat anti-human IgG (γ) heavy chain specific (Boehringer Mannheim 1814249) at 1:10,000 in assay buffer and incubated for 90 min at 25° C. on an orbital shaker. Plates are washed 5× with wash buffer to remove unbound HRP goat anti-human IgG and bound anti-lgG is detected by adding 100 μl of substrate solution (0.4mg/ml o-phenylenedaimine dihydrochloride, Sigma P6912, 6 mM H2O2 in PBS) and incubating for 8 min at 25° C. Enzymatic reaction is stopped by the addition of 100 μl 4.5N H2SO4 and colorimetric product is measured at 490 nm on a 96 well plate densitometer (Molecular Devices). Binding of variant is expressed as a percent of the wild type molecule.
  • FcγRII and III binding ELISAs were performed as described in Example 1 above. [0239]
  • For measuring FcRn binding activity of IgG variants, ELISA plates were coated with 2 μg/ml streptavidin (Zymed, South San Francisco) in 50 mM carbonate buffer, pH 9.6, at 4° C. overnight and blocked with PBS-0.5% BSA, pH 7.2 at room temperature for one hour. Biotinylated FcRn (prepared using biotin-X-NHS from Research Organics, Cleveland, Ohio and used at 1-2 μg/ml) in PBS-0.5% BSA, 0.05[0240] % polysorbate 20, pH 7.2, was added to the plate and incubated for one hour. Two fold serial dilutions of IgG standard (1.6-100 ng/ml) or variants in PBS-0.5% BSA, 0.05% polysorbate 20, pH 6.0, were added to the plate and incubated for two hours. Bound IgG was detected using peroxidase labeled goat F(ab′)2 anti-human lgG F(ab′)2 in the above pH 6.0 buffer (Jackson lmmunoResearch, West Grove, Pa.) followed by 3,3′,5,5′-tetramethyl benzidine (Kirgaard & Perry Laboratories) as the substrate. Plates were washed between steps with PBS-0.05% polysorbate 20 at either pH 7.2 or 6.0. Absorbance was read at 450 nm on a Vmax plate reader (Molecular Devices, Menlo Park, Calif.). Titration curves were fit with a four-parameter nonlinear regression curve-fitfing program (KaleidaGraph, Synergy software, Reading, Pa.). Concentrations of IgG variants corresponding to the mid-point absorbance of the titration curve of the standard were calculated and then divided by the concentration of the standard corresponding to the mid-point absorbance of the standard titration curve.
    • RESULTS AND DISCUSSION
  • By alanine scanning mutagenesis, several single point mutations were constructed in the CH2 domain of C2B8 beginning with E318A, K320A and K322A. All the mutants constructed bound normally to the CD20 antigen (Table 1). [0241]
    TABLE 1
    wt E318A K320A K322A P329A P331A
    FcRn + + + +
    CD20 + + + + + +
    FcγRI + + + + + +
    FcγRII + + + + + +
    FcγRIII + + + + + +
    *C1q +++ ++ +++ − − −
    CDC + + + − − −
  • Where binding of human complement to an antibody with a human Fc was analyzed, the ability of E318A and K320A to activate complement was essentially identical to that of wild type C2B8 (Table 1). When compared to wild type C2B8, there appears to be little difference in the binding of E318A and K320A to C1q. There is only a 10% decrease in the binding of K320A and about a 30% decrease in the binding of E318A to C1q (FIG. 2). The results indicate that the effect of the E318A and the K320A substitution on complement activation and C1q binding is minimal. Also, the human IgG1 of C2B8 was substituted for human IgG2 and used as a negative control in the C1q binding studies. The IgG2 mutant appears to have a much lower affinity for C1q than the E318A and K320A mutants (FIG. 2). Thus, the results demonstrate that E318 and K320 do not constitute the core C1q binding sites for human IgG1. Conversely, the K322A substitution had a significant effect on both complement activity and C1q binding. The K322A mutant did not have CDC activity when tested in the above CDC assay and was more than a 100 fold lower than wild type C2B8 in binding to C1q (FIG. 2). In the human system, K322 is the only residue of the proposed core C1q binding sites that appeared to have a significant effect on complement activation and C1q binding. [0242]
  • Since the Duncan and Winter study was performed using mouse IgG2b and the above results reveal that K320 and E318 in human IgG1 are not involved in C1q binding, and without being bound to any one theory, the above data suggest that the C1q binding region in murine IgGs is different from that of the human. To investigate this further and also to identify additional mutants that do not bind to C1 q and hence do not activate complement, several more point mutations in the vicinity of K322 were constructed as assessed from the three dimensional structure of the C2B8 Fc. Mutants constructed, K274A, N276A, Y278A, S324A, P329A, P331A, K334A, and T335A, were assessed for their ability to bind C1q and also to activate complement. Many of these substitutions had little or no effect on C1q binding or complement activation. In the above assays, the P329 A and the P331 A mutants did not activate complement and had decreased binding to C1q. The P331 A mutant did not activate complement and was 60 fold lower in binding to C1q (FIG. 3) when compared to wild type C2B8 (FIG. 2). The concentration range of the antibody variants used in FIG. 3 is expanded to 100 μg/ml in order to observe saturation of C1q binding to the P331 A variant. The mutation P329A results in an antibody that does not activate complement and is more than a 100 fold lower in binding to C1q (FIG. 3) when compared to wild type C2B8 (FIG. 2). [0243]
  • Mutants that did not bind to C1q and hence did not activate complement were examined for their ability to bind to the Fc receptors: FcγRI, FcγRIIa, FcγRIlb, FcγRIIIa and FcRn. This particular study was performed using a humanized anti-IgE antibody, an IgG1 antibody with these mutations (see Example 1 above). The results revealed the mutants, K322A and P329A, bind to all the Fc receptors to the same extent as the wild type protein (Table 2). However, there was a slight decrease in the binding of P331 A to FcγRIIb. [0244]
  • In conclusion, two amino acid substitutions in the COOH terminal region of the CH2 domain of human IgG1, K322A and P329A were identified that result in more than 100 fold decrease in C1q binding and do not activate the CDC pathway. These two mutants, K322A and P329A, bind to all Fc receptors with the same affinity as the wild type antibody. Based on the results, summarized in Table 2, and without being bound to any one theory, it is proposed that the C1q binding epicenter of human IgG1 is centered around K322, P329 and P331 and is different from the murine IgG2b epicenter which constitutes E318, K320 and K322. [0245]
    TABLE 2
    wt E318A K320A K322A P329A P331A
    CD20
    100 89 102 86 112 103 
    a FcγRI 100 93 102 90 104 74
    a FcγRIIa 100 113 94 109 111 86
    a FcγRIIb 100 106 83 101 96 58
    a FcγIII 100 104 72 90 85 73
    CDC 100 108 108 none none none
  • [0246]
  • 1 2 1 218 PRT Artificial Sequence Artificial Sequence 1-218 Sequence is completely synthesized 1 Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val 1 5 10 15 Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Lys Pro Val Asp 20 25 30 Gly Glu Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln Lys Pro Gly 35 40 45 Lys Ala Pro Lys Leu Leu Ile Tyr Ala Ala Ser Tyr Leu Glu Ser 50 55 60 Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 65 70 75 Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr 80 85 90 Tyr Cys Gln Gln Ser His Glu Asp Pro Tyr Thr Phe Gly Gln Gly 95 100 105 Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe 110 115 120 Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser 125 130 135 Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val 140 145 150 Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 155 160 165 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 170 175 180 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 185 190 195 Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr 200 205 210 Lys Ser Phe Asn Arg Gly Glu Cys 215 218 2 451 PRT Artificial Sequence Artificial Sequence 1-451 Sequence is completely synthesized 2 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly 1 5 10 15 Gly Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Tyr Ser Ile Thr 20 25 30 Ser Gly Tyr Ser Trp Asn Trp Ile Arg Gln Ala Pro Gly Lys Gly 35 40 45 Leu Glu Trp Val Ala Ser Ile Lys Tyr Ser Gly Glu Thr Lys Tyr 50 55 60 Asn Pro Ser Val Lys Gly Arg Ile Thr Ile Ser Arg Asp Asp Ser 65 70 75 Lys Asn Thr Phe Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp 80 85 90 Thr Ala Val Tyr Tyr Cys Ala Arg Gly Ser His Tyr Phe Gly His 95 100 105 Trp His Phe Ala Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser 110 115 120 Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser 125 130 135 Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val 140 145 150 Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 155 160 165 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser 170 175 180 Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 185 190 195 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 200 205 210 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 215 220 225 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 230 235 240 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 260 265 270 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 275 280 285 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 290 295 300 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 305 310 315 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 320 325 330 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 335 340 345 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu 350 355 360 Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly 365 370 375 Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 380 385 390 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 395 400 405 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg 410 415 420 Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 425 430 435 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 440 445 450 Lys 451

Claims (23)

1. A variant of a polypeptide comprising a human IgG Fc region, which variant comprises an amino acid substitution at amino acid position 329, or at two or all of amino acid positions 329, 331 and 322 of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat.
2. The variant of claim 1 having Pro329 of the human IgG Fc region substituted with another amino acid.
3. The variant of claim 1 wherein the polypeptide comprises an antibody.
4. The variant of claim 1 wherein the polypeptide comprises a human IgG1 Fc region.
5. The variant of claim 1 which does not activate complement.
6. The variant of claim 5 which binds an FcR.
7. The variant of claim 6 which binds FcγRI, FcγRII, FcγRIII and FcRn.
8. A variant of a polypeptide comprising a human IgG Fc region, which variant binds FcγRI, FcγRII, FcγRIII and FcRn but does not activate complement and comprises an amino acid substitution at amino acid position 322 or amino acid position 329, or both amino acid positions of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat.
9. A composition comprising the variant of claim 1 and a physiologically acceptable carrier.
10. A method for modifying a polypeptide comprising a human IgG Fc region comprising substituting an amino acid residue at amino acid position 329, or at two or all of amino acid positions 329, 331 and 322 of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat.
11. Isolated nucleic acid encoding the variant of claim 1.
12. A vector comprising the nucleic acid of claim 11.
13. A host cell comprising the vector of claim 12.
14. A process of producing an polypeptide variant comprising culturing the host cell of claim 13 so that the nucleic acid is expressed.
15. The process of claim 14 further comprising recovering the polypeptide variant from the host cell culture.
16. A method of treating a mammal suffering from a disorder comprising administering to the mammal a therapeutically effective amount of a variant of a polypeptide comprising a human IgG Fc region, which variant binds FcγRI, FcγRII, FcγRIII and FcRn but does not activate complement and comprises an amino acid substitution at amino acid position 329, 331 or 322 of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat.
17. An immune complex comprising:
(a) an Fc region-containing polypeptide;
(b) a first target molecule which comprises at least two binding sites for the Fc region-containing polypeptide; and
(c) a second target molecule which comprises at least two binding sites for the first target molecule.
18. A method for determining binding of an analyte to a receptor comprising the following steps performed sequentially:
(a) forming a molecular complex between the analyte and a first target molecule, wherein the first target molecule comprises at least two binding sites for the analyte; and
(b) determining binding of the molecular complex of step (a) to the receptor.
19. The method of claim 18 wherein the analyte is an Fc region-containing polypeptide.
20. The method of claim 18 wherein the receptor is a low affinity receptor.
21. The method of claim 18 wherein the receptor is an FcR.
22. The method of claim 18 wherein the molecular complex of step (a) further comprises a second target molecule which comprises at least two binding sites for the first target molecule.
23. A kit useful for determining binding of an analyte to a receptor comprising:
(a) a first target molecule which comprises at least two binding sites for the analyte; and
(b) a second target molecule which comprises at least two binding sites for the first target molecule.
US09/792,938 1998-04-02 2001-02-26 Polypeptide variants Abandoned US20030166868A1 (en)

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Cited By (55)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030158389A1 (en) * 1998-04-02 2003-08-21 Genentech, Inc. Polypeptide variants
US20040132101A1 (en) * 2002-09-27 2004-07-08 Xencor Optimized Fc variants and methods for their generation
US20040191244A1 (en) * 1999-01-15 2004-09-30 Genentech, Inc. Polypeptide variants with altered effector function
US20050069521A1 (en) * 2003-08-28 2005-03-31 Emd Lexigen Research Center Corp. Enhancing the circulating half-life of interleukin-2 proteins
US20050164352A1 (en) * 2003-12-30 2005-07-28 Emd Lexigen Research Center Corp. IL-7 fusion proteins
WO2005070967A2 (en) * 2004-01-22 2005-08-04 Merck Patent Gmbh Anti-cancer antibodies with reduced complement fixation
US20050244403A1 (en) * 2004-03-24 2005-11-03 Xencor, Inc. Immunoglobulin variants outside the Fc region
US20050249723A1 (en) * 2003-12-22 2005-11-10 Xencor, Inc. Fc polypeptides with novel Fc ligand binding sites
US20060074225A1 (en) * 2004-09-14 2006-04-06 Xencor, Inc. Monomeric immunoglobulin Fc domains
US20060173170A1 (en) * 2004-11-12 2006-08-03 Xencor, Inc. Fc variants with altered binding to FcRn
US20060194290A1 (en) * 1999-01-15 2006-08-31 Genentech, Inc. Polypeptide variants with altered effector function
US20060235208A1 (en) * 2002-09-27 2006-10-19 Xencor, Inc. Fc variants with optimized properties
US20060275283A1 (en) * 2003-11-12 2006-12-07 Biogen Idec Ma Inc. Fcgamma receptor-binding polypeptide variants and methods related thereto
US20060275282A1 (en) * 2005-01-12 2006-12-07 Xencor, Inc. Antibodies and Fc fusion proteins with altered immunogenicity
US20070003546A1 (en) * 2002-03-01 2007-01-04 Xencor, Inc. Optimized Fc variants and methods for their generation
US7183387B1 (en) 1999-01-15 2007-02-27 Genentech, Inc. Polypeptide variants with altered effector function
US20070148171A1 (en) * 2002-09-27 2007-06-28 Xencor, Inc. Optimized anti-CD30 antibodies
US20070219133A1 (en) * 2002-03-01 2007-09-20 Xencor, Inc. CD52 OPTIMIZED Fc VARIANTS AND METHODS FOR THEIR GENERATION
US20070238665A1 (en) * 2003-03-03 2007-10-11 Xencor, Inc. Fc Variants Having Decreased Affinity for FcyRIIc
US20070275460A1 (en) * 2003-03-03 2007-11-29 Xencor.Inc. Fc Variants With Optimized Fc Receptor Binding Properties
US20080057056A1 (en) * 2003-03-03 2008-03-06 Xencor, Inc. Fc Variants with Increased Affinity for FcyRIIC
US20080206867A1 (en) * 2005-10-03 2008-08-28 Desjarlais John R Fc variants with optimized Fc receptor binding properties
US20080254027A1 (en) * 2002-03-01 2008-10-16 Bernett Matthew J Optimized CD5 antibodies and methods of using the same
US20080260731A1 (en) * 2002-03-01 2008-10-23 Bernett Matthew J Optimized antibodies that target cd19
US20080267976A1 (en) * 2005-10-06 2008-10-30 Gregory Alan Lazar Optimized Anti-Cd30 Antibodies
US20080313379A1 (en) * 2007-06-15 2008-12-18 United Memories, Inc. Multiple bus charge sharing
US20090136936A1 (en) * 2007-05-01 2009-05-28 George Georgiou Immunoglobulin fc libraries
US20100104557A1 (en) * 2006-09-18 2010-04-29 Xencor, Inc. Optimized Antibodies that Target HM1.24
US7820161B1 (en) 1999-05-07 2010-10-26 Biogen Idec, Inc. Treatment of autoimmune diseases
US20100272723A1 (en) * 2006-08-14 2010-10-28 Xencor, Inc. Optimized Antibodies that Target CD19
US20100330076A1 (en) * 2009-06-30 2010-12-30 George Georgiou Immunoglobulin fc polypeptides
US8084582B2 (en) 2003-03-03 2011-12-27 Xencor, Inc. Optimized anti-CD20 monoclonal antibodies having Fc variants
US8101720B2 (en) 2004-10-21 2012-01-24 Xencor, Inc. Immunoglobulin insertions, deletions and substitutions
US8188231B2 (en) 2002-09-27 2012-05-29 Xencor, Inc. Optimized FC variants
US8318907B2 (en) 2004-11-12 2012-11-27 Xencor, Inc. Fc variants with altered binding to FcRn
US8409577B2 (en) 2006-06-12 2013-04-02 Emergent Product Development Seattle, Llc Single chain multivalent binding proteins with effector function
US8546543B2 (en) 2004-11-12 2013-10-01 Xencor, Inc. Fc variants that extend antibody half-life
US8802820B2 (en) 2004-11-12 2014-08-12 Xencor, Inc. Fc variants with altered binding to FcRn
US8853366B2 (en) 2001-01-17 2014-10-07 Emergent Product Development Seattle, Llc Binding domain-immunoglobulin fusion proteins
US8969526B2 (en) 2011-03-29 2015-03-03 Roche Glycart Ag Antibody Fc variants
US9051373B2 (en) 2003-05-02 2015-06-09 Xencor, Inc. Optimized Fc variants
US9101609B2 (en) 2008-04-11 2015-08-11 Emergent Product Development Seattle, Llc CD37 immunotherapeutic and combination with bifunctional chemotherapeutic thereof
US9238878B2 (en) 2009-02-17 2016-01-19 Redwood Bioscience, Inc. Aldehyde-tagged protein-based drug carriers and methods of use
US9475881B2 (en) 2010-01-19 2016-10-25 Xencor, Inc. Antibody variants with enhanced complement activity
US9540438B2 (en) 2011-01-14 2017-01-10 Redwood Bioscience, Inc. Aldehyde-tagged immunoglobulin polypeptides and methods of use thereof
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US9714282B2 (en) 2003-09-26 2017-07-25 Xencor, Inc. Optimized Fc variants and methods for their generation
US10143748B2 (en) 2005-07-25 2018-12-04 Aptevo Research And Development Llc B-cell reduction using CD37-specific and CD20-specific binding molecules
US10457737B2 (en) 2015-02-09 2019-10-29 Research Development Foundation Engineered immunoglobulin Fc polypeptides displaying improved complement activation
US10654916B2 (en) 2011-04-21 2020-05-19 The Regents Of The University Of California, A California Corporation Compositions and methods for the treatment of neuromyelitis optica
US11208632B2 (en) 2016-04-26 2021-12-28 R.P. Scherer Technologies, Llc Antibody conjugates and methods of making and using the same
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US11401348B2 (en) 2009-09-02 2022-08-02 Xencor, Inc. Heterodimeric Fc variants
US11820830B2 (en) 2004-07-20 2023-11-21 Xencor, Inc. Optimized Fc variants
US11932685B2 (en) 2007-10-31 2024-03-19 Xencor, Inc. Fc variants with altered binding to FcRn

Families Citing this family (1531)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TWI239847B (en) * 1997-12-02 2005-09-21 Elan Pharm Inc N-terminal fragment of Abeta peptide and an adjuvant for preventing and treating amyloidogenic disease
US7179892B2 (en) * 2000-12-06 2007-02-20 Neuralab Limited Humanized antibodies that recognize beta amyloid peptide
US20080050367A1 (en) * 1998-04-07 2008-02-28 Guriq Basi Humanized antibodies that recognize beta amyloid peptide
US7964192B1 (en) 1997-12-02 2011-06-21 Janssen Alzheimer Immunotherapy Prevention and treatment of amyloidgenic disease
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US6761888B1 (en) * 2000-05-26 2004-07-13 Neuralab Limited Passive immunization treatment of Alzheimer's disease
US6194551B1 (en) * 1998-04-02 2001-02-27 Genentech, Inc. Polypeptide variants
GB9809951D0 (en) 1998-05-08 1998-07-08 Univ Cambridge Tech Binding molecules
WO2001060397A1 (en) * 2000-02-16 2001-08-23 Genentech, Inc. Uses of agonists and antagonists to modulate activity of tnf-related molecules
CN1373672A (en) * 1999-07-12 2002-10-09 杰南技术公司 Blocking immune response to foreign antigen using antagonist which binds to CD20
JP4992068B2 (en) * 2000-01-27 2012-08-08 メディミューン，エルエルシー Ultra high affinity neutralizing antibody
EP1259547B1 (en) 2000-03-01 2012-07-11 Medimmune, Inc. High potency recombinant antibodies and method for producing them
US7416726B2 (en) 2000-04-13 2008-08-26 The Rockefeller University Enhancement of antibody-mediated immune responses
KR20040023565A (en) * 2000-09-18 2004-03-18 아이덱 파마슈티칼즈 코포레이션 Combination therapy for treatment of autoimmune diseases using b cell depleting/immunoregulatory antibody combination
US6787524B2 (en) * 2000-09-22 2004-09-07 Tanox, Inc. CpG oligonucleotides and related compounds for enhancing ADCC induced by anti-IgE antibodies
US7179900B2 (en) * 2000-11-28 2007-02-20 Medimmune, Inc. Methods of administering/dosing anti-RSV antibodies for prophylaxis and treatment
US6855493B2 (en) 2000-11-28 2005-02-15 Medimmune, Inc. Methods of administering/dosing anti-RSV antibodies for prophylaxis and treatment
TWI255272B (en) * 2000-12-06 2006-05-21 Guriq Basi Humanized antibodies that recognize beta amyloid peptide
US7700751B2 (en) 2000-12-06 2010-04-20 Janssen Alzheimer Immunotherapy Humanized antibodies that recognize β-amyloid peptide
CA2431600C (en) * 2000-12-12 2012-04-17 Medimmune, Inc. Molecules with extended half-lives, compositions and uses thereof
US7658921B2 (en) * 2000-12-12 2010-02-09 Medimmune, Llc Molecules with extended half-lives, compositions and uses thereof
US7829084B2 (en) * 2001-01-17 2010-11-09 Trubion Pharmaceuticals, Inc. Binding constructs and methods for use thereof
US20030133939A1 (en) * 2001-01-17 2003-07-17 Genecraft, Inc. Binding domain-immunoglobulin fusion proteins
US20040058445A1 (en) * 2001-04-26 2004-03-25 Ledbetter Jeffrey Alan Activation of tumor-reactive lymphocytes via antibodies or genes recognizing CD3 or 4-1BB
JP2005510208A (en) * 2001-08-03 2005-04-21 ジェネンテック・インコーポレーテッド TACIs and BR3 polypeptides and uses thereof
US20060073095A1 (en) * 2001-08-22 2006-04-06 Steven Kessler Methods for screening antibody-producing cells on heterogeneous antigen substrates
US20040235068A1 (en) * 2001-09-05 2004-11-25 Levinson Arthur D. Methods for the identification of polypeptide antigens associated with disorders involving aberrant cell proliferation and compositions useful for the treatment of such disorders
US20110045005A1 (en) 2001-10-19 2011-02-24 Craig Crowley Compositions and methods for the treatment of tumor of hematopoietic origin
JP2005532253A (en) * 2001-10-25 2005-10-27 ジェネンテック・インコーポレーテッド Glycoprotein composition
US6911321B2 (en) * 2001-12-19 2005-06-28 Genentech, Inc. Non-human primate Fc receptors and methods of use
WO2003068801A2 (en) * 2002-02-11 2003-08-21 Genentech, Inc. Antibody variants with faster antigen association rates
US20040002587A1 (en) * 2002-02-20 2004-01-01 Watkins Jeffry D. Fc region variants
US20100311954A1 (en) * 2002-03-01 2010-12-09 Xencor, Inc. Optimized Proteins that Target Ep-CAM
US20090042291A1 (en) * 2002-03-01 2009-02-12 Xencor, Inc. Optimized Fc variants
MY139983A (en) * 2002-03-12 2009-11-30 Janssen Alzheimer Immunotherap Humanized antibodies that recognize beta amyloid peptide
US20030219436A1 (en) * 2002-03-15 2003-11-27 Ledbetter Jeffrey A. Compositions and methods to regulate an immune response using CD83 gene expressed in tumors and using soluble CD83-Ig fusion protein
EP1513554B9 (en) * 2002-05-30 2011-11-09 Macrogenics, Inc. Cd16a binding proteins and use for the treatment of immune disorders
US7132100B2 (en) 2002-06-14 2006-11-07 Medimmune, Inc. Stabilized liquid anti-RSV antibody formulations
JP2005533863A (en) * 2002-07-25 2005-11-10 ジェネンテック・インコーポレーテッド TACI antibodies and their uses
US8187593B2 (en) * 2002-08-14 2012-05-29 Macrogenics, Inc. FcγRIIB specific antibodies and methods of use thereof
US8946387B2 (en) * 2002-08-14 2015-02-03 Macrogenics, Inc. FcγRIIB specific antibodies and methods of use thereof
US8530627B2 (en) * 2002-08-14 2013-09-10 Macrogenics, Inc. FcγRIIB specific antibodies and methods of use thereof
US8968730B2 (en) 2002-08-14 2015-03-03 Macrogenics Inc. FcγRIIB specific antibodies and methods of use thereof
DK1534335T4 (en) * 2002-08-14 2015-10-05 Macrogenics Inc FCGAMMARIIB-SPECIFIC ANTIBODIES AND PROCEDURES FOR USE THEREOF
US8193318B2 (en) * 2002-08-14 2012-06-05 Macrogenics, Inc. FcγRIIB specific antibodies and methods of use thereof
US8044180B2 (en) * 2002-08-14 2011-10-25 Macrogenics, Inc. FcγRIIB specific antibodies and methods of use thereof
EP2364996B1 (en) 2002-09-27 2016-11-09 Xencor Inc. Optimized FC variants and methods for their generation
NZ568769A (en) 2002-10-17 2010-04-30 Genmab As Human monoclonal antibodies against CD20
DE10254601A1 (en) 2002-11-22 2004-06-03 Ganymed Pharmaceuticals Ag Gene products differentially expressed in tumors and their use
BRPI0316779B8 (en) * 2002-12-16 2023-02-28 Genentech Inc HUMAN ANTI-CD20 ANTIBODY OR ANTIGEN-BINDING FRAGMENT THEREOF, ITS USES, COMPOSITION, MANUFACTURED ARTICLE AND LIQUID FORMULATION
EP2368578A1 (en) * 2003-01-09 2011-09-28 Macrogenics, Inc. Identification and engineering of antibodies with variant Fc regions and methods of using same
US7960512B2 (en) 2003-01-09 2011-06-14 Macrogenics, Inc. Identification and engineering of antibodies with variant Fc regions and methods of using same
EP2272533A1 (en) 2003-01-13 2011-01-12 MacroGenics, Inc. Soluble FcyR fusion proteins and methods of use thereof
MXPA05008156A (en) * 2003-02-01 2005-09-30 Neuralab Ltd Active immunization to generate antibodies to soluble a-beta.
JP4928259B2 (en) 2003-03-04 2012-05-09 キリンホールディングス株式会社 Endothelial cell-specific antibody and use thereof
JP2006522811A (en) * 2003-04-09 2006-10-05 ジェネンテック・インコーポレーテッド Method for treating autoimmune disease in patients with inadequate response to TNFα inhibitors
TWI353991B (en) 2003-05-06 2011-12-11 Syntonix Pharmaceuticals Inc Immunoglobulin chimeric monomer-dimer hybrids
EP1626993B1 (en) * 2003-05-09 2015-03-11 Duke University Cd20-specific antibodies and methods of employing same
TWI374893B (en) * 2003-05-30 2012-10-21 Janssen Alzheimer Immunotherap Humanized antibodies that recognize beta amyloid peptide
JP2007526220A (en) * 2003-06-05 2007-09-13 ジェネンテック・インコーポレーテッド Combination therapy for B cell disease
US20050163775A1 (en) * 2003-06-05 2005-07-28 Genentech, Inc. Combination therapy for B cell disorders
US20050232931A1 (en) * 2003-06-13 2005-10-20 Oncomax Acquisition Corp. Preparation and application of anti-tumor bifunctional fusion proteins
US20040254108A1 (en) * 2003-06-13 2004-12-16 Jing Ma Preparation and application of anti-tumor bifunctional fusion proteins
WO2005003175A2 (en) * 2003-06-13 2005-01-13 Biogen Idec Ma Inc. Aglycosyl anti-cd154 (cd40 ligand) antibodies and uses thereof
CN104645327A (en) 2003-07-24 2015-05-27 依奈特制药公司 Methods and compositions for increasing the efficiency of therapeutic antibodies using NK cell potentiating compounds
US7754209B2 (en) 2003-07-26 2010-07-13 Trubion Pharmaceuticals Binding constructs and methods for use thereof
JP2007500844A (en) * 2003-07-29 2007-01-18 ジェネンテック・インコーポレーテッド Human anti-CD20 antibody assays and uses
EP1664116A4 (en) 2003-08-22 2009-06-03 Biogen Idec Inc Improved antibodies having altered effector function and methods for making the same
WO2005023302A2 (en) * 2003-08-29 2005-03-17 Genentech, Inc. Anti-cd20 therapy of ocular disorders
TWI333977B (en) * 2003-09-18 2010-12-01 Symphogen As Method for linking sequences of interest
RS53984B1 (en) 2003-12-10 2015-10-30 E. R. Squibb & Sons L.L.C. Ip-10 antibodies and their uses
SI1711207T1 (en) 2003-12-10 2013-07-31 Medarex, L.L.C Interferon alpha antibodies and their uses
KR20060107555A (en) * 2003-12-19 2006-10-13 제넨테크, 인크. Detection of cd20 in transplant rejection
CA2549122A1 (en) * 2003-12-19 2005-07-07 Genentech, Inc. Detection of cd20 in therapy of autoimmune diseases
US20090324604A1 (en) 2004-01-07 2009-12-31 Chiron Corporation M-csf specific monoclonal antibody and uses thereof
DK1716181T3 (en) 2004-02-19 2010-03-01 Genentech Inc CDR repaired antibodies
AR049021A1 (en) * 2004-04-16 2006-06-21 Genentech Inc TREATMENT OF DISORDERS WITH AN ANTIBODY THAT JOINS CD20
KR20070012408A (en) * 2004-04-16 2007-01-25 제넨테크, 인크. Method for augmenting b cell depletion
JP5367982B2 (en) * 2004-04-16 2013-12-11 マクロジェニクス，インコーポレーテッド FcγRIIB specific antibody and method of use thereof
MXPA06012674A (en) * 2004-05-05 2007-03-26 Genentech Inc Preventing autoimmune disease.
EP1761563A4 (en) * 2004-05-10 2008-05-14 Macrogenics Inc HUMANIZED Fc gamma RIIB SPECIFIC ANTIBODIES AND METHODS OF USE THEREOF
DE102004024617A1 (en) 2004-05-18 2005-12-29 Ganymed Pharmaceuticals Ag Differentially expressed in tumors gene products and their use
PL1753871T3 (en) 2004-05-28 2016-01-29 Agensys Inc Antibodies and related molecules that bind to psca proteins
KR20070029733A (en) 2004-06-04 2007-03-14 제넨테크, 인크. Method for treating multiple sclerosis
AU2005251764A1 (en) * 2004-06-04 2005-12-22 Genentech, Inc. Method for treating lupus
CA2571218C (en) 2004-06-17 2015-11-03 William D. Carlson Tdf-related compounds and analogs thereof
PL2662390T3 (en) 2004-06-21 2017-12-29 E. R. Squibb & Sons, L.L.C. Interferon alpha receptor 1 antibodies and their uses
ES2530340T3 (en) 2004-07-15 2015-03-02 Xencor Inc Optimized Fc variants
EP1781378A2 (en) * 2004-07-22 2007-05-09 Genentech, Inc. Method of treating sjögren's syndrome
WO2006033702A2 (en) 2004-07-26 2006-03-30 Biogen Idec Ma Inc. Anti-cd154 antibodies
CA2575607C (en) 2004-08-03 2017-07-11 Innate Pharma Therapeutic and diagnostic methods and compositions targeting 4ig-b7-h3 and its counterpart nk cell receptor
CA2573192C (en) 2004-08-04 2013-09-10 Applied Molecular Evolution Inc. Variant fc regions
US7659374B2 (en) * 2004-08-16 2010-02-09 Medimmune, Llc Eph receptor Fc variants with enhanced antibody dependent cell-mediated cytotoxicity activity
BRPI0515230A (en) * 2004-08-19 2008-07-15 Genentech Inc isolated polypeptides, antibodies and nucleic acids, compositions, expression vector, isolated host cells, method of producing an antibody, manufactured articles, methods of treatment and alleviating dysfunction, methods of making and selecting a polypeptide, antibody cd20 binding agent, isolated anti-her2 antibody and uses of an antibody
KR20070100228A (en) * 2004-10-05 2007-10-10 제넨테크, 인크. Method for treating vasculitis
JO3000B1 (en) 2004-10-20 2016-09-05 Genentech Inc Antibody Formulations.
EP1812068A4 (en) * 2004-10-29 2010-06-09 Medimmune Inc Methods of preventing and treating rsv infections and related conditions
AU2005335714B2 (en) 2004-11-10 2012-07-26 Macrogenics, Inc. Engineering Fc antibody regions to confer effector function
US20060222653A1 (en) * 2004-11-12 2006-10-05 Xencor, Inc. Antibodies operably linked to selected chemoattractants
US20070135620A1 (en) * 2004-11-12 2007-06-14 Xencor, Inc. Fc variants with altered binding to FcRn
PE20061401A1 (en) * 2004-12-15 2006-12-23 Neuralab Ltd Aß ANTIBODIES TO IMPROVE COGNITION
ES2396555T3 (en) * 2004-12-15 2013-02-22 Janssen Alzheimer Immunotherapy Antibodies that recognize beta amyloid peptide
WO2006066089A1 (en) * 2004-12-15 2006-06-22 Neuralab Limited Humanized amyloid beta antibodies for use in improving cognition
EP1674479A1 (en) * 2004-12-22 2006-06-28 Memorial Sloan-Kettering Cancer Center Modulation of Fc Gamma receptors for optimizing immunotherapy
WO2006076651A2 (en) * 2005-01-13 2006-07-20 Genentech, Inc. Treatment method
KR20130089280A (en) 2005-01-21 2013-08-09 제넨테크, 인크. Fixed dosing of her antibodies
DOP2006000029A (en) * 2005-02-07 2006-08-15 Genentech Inc ANTIBODY VARIANTS AND USES THEREOF. (VARIATIONS OF AN ANTIBODY AND USES OF THE SAME)
CA2597924C (en) 2005-02-15 2018-10-02 Duke University Anti-cd19 antibodies and uses in oncology
US20060263357A1 (en) 2005-05-05 2006-11-23 Tedder Thomas F Anti-CD19 antibody therapy for autoimmune disease
RU2404806C2 (en) 2005-02-23 2010-11-27 Дженентек, Инк. Extension of time to progression of disease or lifetime of oncologic patients with application of her dimerisation inhibitors
TW200714289A (en) * 2005-02-28 2007-04-16 Genentech Inc Treatment of bone disorders
US20060204505A1 (en) * 2005-03-08 2006-09-14 Sliwkowski Mark X Methods for identifying tumors responsive to treatment with HER dimerization inhibitors (HDIs)
NZ561883A (en) 2005-03-23 2010-11-26 Genmab As Antibodies against CD38 for treatment of multiple myeloma
RU2413735C2 (en) 2005-03-31 2011-03-10 Эдженсис, Инк. Antibodies and related molecules binding with proteins 161p2f10b
ES2971647T3 (en) 2005-04-15 2024-06-06 Macrogenics Inc Covalent diabodies and their uses
US9284375B2 (en) 2005-04-15 2016-03-15 Macrogenics, Inc. Covalent diabodies and uses thereof
AR053579A1 (en) * 2005-04-15 2007-05-09 Genentech Inc TREATMENT OF INTESTINAL INFLAMMATORY DISEASE (IBD)
US11254748B2 (en) 2005-04-15 2022-02-22 Macrogenics, Inc. Covalent diabodies and uses thereof
US9963510B2 (en) 2005-04-15 2018-05-08 Macrogenics, Inc. Covalent diabodies and uses thereof
WO2010080538A1 (en) 2008-12-19 2010-07-15 Macrogenics, Inc. Covalent diabodies and uses thereof
US20060240007A1 (en) * 2005-04-22 2006-10-26 Genentech, Inc. Method for treating dementia or Alzheimer's disease
CA2606102C (en) 2005-04-26 2014-09-30 Medimmune, Inc. Modulation of antibody effector function by hinge domain engineering
EP3530736A3 (en) 2005-05-09 2019-11-06 ONO Pharmaceutical Co., Ltd. Human monoclonal antibodies to programmed death 1 (pd-1) and methods for treating cancer using anti-pd-1 antibodies alone or in combination with other immunotherapeutics
CN101228189A (en) * 2005-05-09 2008-07-23 格黎卡特生物技术股份公司 Antigen binding molecules having modified FC regions and altered binding to FC receptors
US7601335B2 (en) * 2005-05-20 2009-10-13 Genentech, Inc. Pretreatment of a biological sample from an autoimmune disease subject
EP1893647A2 (en) 2005-06-23 2008-03-05 MedImmune, Inc. Antibody formulations having optimized aggregation and fragmentation profiles
JP2009500344A (en) * 2005-07-01 2009-01-08 メディミューン，エルエルシー An integrated approach to manufacturing multidomain protein therapeutics
WO2007005874A2 (en) 2005-07-01 2007-01-11 Medarex, Inc. Human monoclonal antibodies to programmed death ligand 1 (pd-l1)
WO2007008943A2 (en) 2005-07-08 2007-01-18 Xencor, Inc. Optimized anti-ep-cam antibodies
KR20080047540A (en) 2005-07-25 2008-05-29 트루비온 파마슈티칼스, 인코포레이티드 Single dose use of cd20-specific binding molecules
US20080279850A1 (en) * 2005-07-25 2008-11-13 Trubion Pharmaceuticals, Inc. B-Cell Reduction Using CD37-Specific and CD20-Specific Binding Molecules
ES2538036T3 (en) * 2005-07-28 2015-06-16 Novartis Ag Use of antibody to M-CSF
JP5457671B2 (en) * 2005-07-28 2014-04-02 ノバルティス アーゲー M-CSF specific monoclonal antibody and use thereof
PT2573114T (en) 2005-08-10 2016-07-13 Macrogenics Inc Identification and engineering of antibodies with variant fc regions and methods of using same
EP1762575A1 (en) 2005-09-12 2007-03-14 Ganymed Pharmaceuticals AG Identification of tumor-associated antigens for diagnosis and therapy
US8299212B2 (en) 2005-09-20 2012-10-30 Thrasos Therapeutics, Inc. TDF-related compounds and analogs thereof, analogs and bioactive fragments
CA2625619A1 (en) 2005-10-14 2007-04-26 Medimmune, Inc. Cell display of antibody libraries
PT2532679T (en) 2005-10-21 2017-07-18 Novartis Ag Human antibodies against il13 and therapeutic uses
US20070099246A1 (en) * 2005-11-03 2007-05-03 Sandy John D Antibodies, assays and kits to quantitate cartilage destruction
MY149159A (en) 2005-11-15 2013-07-31 Hoffmann La Roche Method for treating joint damage
JP6088723B2 (en) * 2005-11-23 2017-03-01 ジェネンテック， インコーポレイテッド Compositions and methods for B cell assays.
EP1790664A1 (en) 2005-11-24 2007-05-30 Ganymed Pharmaceuticals AG Monoclonal antibodies against claudin-18 for treatment of cancer
CN105859886A (en) 2005-12-02 2016-08-17 健泰科生物技术公司 Compositions and methods associated with antibodies that bind to IL-22 and IL-22R
US8383118B2 (en) 2005-12-08 2013-02-26 Medarex, Inc. Human monoclonal antibodies to fucosyl-GM1 and methods for using anti-fucosyl-GM1
EP2402373B1 (en) 2006-01-05 2016-10-19 Genentech, Inc. Anti-EphB4 Antibodies and Methods Using Same
AU2007207764B2 (en) 2006-01-12 2012-06-07 Alexion Pharmaceuticals, Inc. Antibodies to OX-2/CD200 and uses thereof
CA2638133A1 (en) 2006-01-20 2007-11-08 Genentech, Inc. Anti-ephrinb2 antibodies and methods using same
JP2009525986A (en) * 2006-02-03 2009-07-16 メディミューン，エルエルシー Protein preparation
EP2540741A1 (en) 2006-03-06 2013-01-02 Aeres Biomedical Limited Humanized anti-CD22 antibodies and their use in treatment of oncology, transplantation and autoimmune disease
EP1999470A4 (en) * 2006-03-10 2009-08-19 Macrogenics Inc Identification and engineering of antibodies with variant heavy chains and methods of using same
AR059851A1 (en) 2006-03-16 2008-04-30 Genentech Inc ANTIBODIES OF EGFL7 AND METHODS OF USE
WO2007111661A2 (en) 2006-03-20 2007-10-04 Xoma Technology Ltd. Human antibodies specific for gastrin materials and methods
RS54156B1 (en) 2006-03-21 2015-12-31 Genentech Inc. Combinatorial therapy involving alpha5beta1 antagonists
US8784810B2 (en) * 2006-04-18 2014-07-22 Janssen Alzheimer Immunotherapy Treatment of amyloidogenic diseases
WO2009017467A1 (en) 2007-07-27 2009-02-05 Elan Pharma International Limited Treatment of amyloidogenic diseases
ES2489646T3 (en) * 2006-05-26 2014-09-02 Macrogenics, Inc. Humanized antibodies specific to Fc gamma RIIB and its methods of use
PL2446904T3 (en) 2006-05-30 2015-10-30 Genentech Inc Anti-CD22 antibodies, their immunoconjugates and uses thereof
KR20090027227A (en) * 2006-06-06 2009-03-16 제넨테크, 인크. Anti-dll4 antibodies and methods using same
CL2007001624A1 (en) * 2006-06-06 2008-01-18 Genentech Inc Use of an antibody that binds to dll4 for the treatment of tumor, cancer or cell proliferation disorder.
EP2029173B1 (en) 2006-06-26 2016-07-20 MacroGenics, Inc. Fc riib-specific antibodies and methods of use thereof
WO2008002933A2 (en) 2006-06-26 2008-01-03 Macrogenics, Inc. Combination of fcgammariib antibodies and cd20-specific antibodies and methods of use thereof
AU2007275654A1 (en) 2006-07-19 2008-01-24 The Trustees Of The University Of Pennsylvania WSX-1/p28 as a target for anti-inflammatory responses
MX2009001341A (en) * 2006-08-04 2009-05-28 Novartis Ag Ephb3-specific antibody and uses thereof.
PT2059535E (en) 2006-08-18 2014-01-29 Novartis Ag Prlr-specific antibody and uses thereof
BRPI0714893A2 (en) 2006-09-05 2013-05-28 Medarex Inc isolated monoclonal antibody or antigen-binding portion thereof, antibody fragment, mimetic antibody, immunoconjugate, isolated nucleic acid molecule composition, expression vector, host cell, method for preparing an anti-bmp2 or anti-bmp4 antibody, Method for treating or preventing a disease associated with normal bone formation and ossification, hybridoma and method for preparing the antibody
PL2066349T3 (en) 2006-09-08 2012-09-28 Medimmune Llc Humanized anti-cd19 antibodies and their use in treatment of tumors, transplantation and autoimmune diseases
CA2662236A1 (en) 2006-09-12 2008-03-20 Genentech, Inc. Methods and compositions for the diagnosis and treatment of cancer
ME02269B (en) 2006-10-02 2016-04-28 Medarex Inc Human antibodies that bind cxcr4 and uses thereof
US20080112961A1 (en) * 2006-10-09 2008-05-15 Macrogenics, Inc. Identification and Engineering of Antibodies with Variant Fc Regions and Methods of Using Same
AU2007319605B2 (en) 2006-10-19 2011-02-17 Csl Limited Anti-IL-13R alpha 1 antibodies and their uses thereof
PL2068921T3 (en) 2006-10-19 2014-12-31 Csl Ltd High affinity antibody antagonists of interleukin-13 receptor alpha 1
AU2007308792B2 (en) 2006-10-27 2013-01-24 Genentech, Inc. Antibodies and immunoconjugates and uses therefor
US8618248B2 (en) 2006-10-31 2013-12-31 President And Fellows Of Harvard College Phosphopeptide compositions and anti-phosphopeptide antibody compositions and methods of detecting phosphorylated peptides
JP2010508816A (en) 2006-11-02 2010-03-25 アクセルロン ファーマ， インコーポレイテッド ALK1 receptor and ligand antagonist and use thereof
US8247537B2 (en) 2006-11-15 2012-08-21 Medarex, Inc. Human monoclonal antibodies to BTLA and methods of use
ATE555128T1 (en) * 2006-11-30 2012-05-15 Res Dev Foundation IMPROVED IMMUNOLOBULIN LIBRARIES
NZ578064A (en) 2006-12-01 2012-01-12 Medarex Inc Human antibodies that bind cd22 and uses thereof
KR101519672B1 (en) 2006-12-07 2015-05-29 노파르티스 아게 Antagonist antibodies against ephb3
WO2008140603A2 (en) 2006-12-08 2008-11-20 Macrogenics, Inc. METHODS FOR THE TREATMENT OF DISEASE USING IMMUNOGLOBULINS HAVING FC REGIONS WITH ALTERED AFFINITIES FOR FCγR ACTIVATING AND FCγR INHIBITING
CL2007003622A1 (en) 2006-12-13 2009-08-07 Medarex Inc Human anti-cd19 monoclonal antibody; composition comprising it; and tumor cell growth inhibition method.
AU2007333098A1 (en) 2006-12-14 2008-06-19 Medarex, Inc. Human antibodies that bind CD70 and uses thereof
US9127064B2 (en) 2006-12-21 2015-09-08 Novo Nordisk A/S Antibodies against human NKG2D and uses thereof
AR064464A1 (en) * 2006-12-22 2009-04-01 Genentech Inc ANTIBODIES ANTI - INSULINAL GROWTH FACTOR RECEIVER
US20100119526A1 (en) * 2007-01-26 2010-05-13 Bioinvent International Ab DLL4 Signaling Inhibitors and Uses Thereof
KR20090114443A (en) 2007-02-09 2009-11-03 제넨테크, 인크. Anti-Robo4 Antibodies and Uses Therefor
ATE521704T1 (en) * 2007-03-01 2011-09-15 Symphogen As METHOD FOR CLONING RELATED ANTIBODIES
DK2132573T3 (en) 2007-03-02 2014-07-14 Genentech Inc PREDICTION OF RESPONSE TO A HER DIMERIZATION INHIBITOR BASED ON LOW HER3 EXPRESSION
JP5311060B2 (en) 2007-03-08 2013-10-09 カロバイオス ファーマシューティカルズ インコーポレイティッド EphA3 antibody for the treatment of solid tumors
EP1970384A1 (en) 2007-03-14 2008-09-17 Ganymed Pharmaceuticals AG Monoclonal antibodies for treatment of cancer
MY166021A (en) 2007-03-22 2018-05-21 Biogen Ma Inc Binding proteins,including antibodies,antibody derivatives and antibody fragments,that specifically bind cd154 and uses thereof
WO2008118324A2 (en) 2007-03-26 2008-10-02 Macrogenics, Inc. Composition and method of treating cancer with an anti-uroplakin ib antibody
US8003097B2 (en) * 2007-04-18 2011-08-23 Janssen Alzheimer Immunotherapy Treatment of cerebral amyloid angiopathy
US20080292625A1 (en) * 2007-04-18 2008-11-27 Sally Schroeter Prevention and treatment of cerebral amyloid angiopathy
US20130195881A1 (en) 2007-04-27 2013-08-01 Sanjaya Singh Potent, stable and non-immunosuppressive anti-cd4 antibodies
KR20100017514A (en) 2007-05-07 2010-02-16 메디뮨 엘엘씨 Anti-icos antibodies and their use in treatment of oncology, transplantation and autoimmune disease
EP1997832A1 (en) 2007-05-29 2008-12-03 Ganymed Pharmaceuticals AG Monoclonal antibodies against Claudin-18 for treatment of cancer
EP1997830A1 (en) * 2007-06-01 2008-12-03 AIMM Therapeutics B.V. RSV specific binding molecules and means for producing them
PE20090321A1 (en) 2007-06-04 2009-04-20 Genentech Inc ANTI-NOTCH1 NRR ANTIBODIES, METHOD OF PREPARATION AND PHARMACEUTICAL COMPOSITION
DK2171090T3 (en) 2007-06-08 2013-06-10 Genentech Inc Gene expression markers for tumor resistance to HER2 inhibitor therapy
KR101599735B1 (en) 2007-06-21 2016-03-07 마크로제닉스, 인크. Covalent diabodies and uses thereof
US20110077383A1 (en) * 2007-07-03 2011-03-31 Medimmune, Llc Hinge domain engineering
KR102055873B1 (en) 2007-07-09 2019-12-13 제넨테크, 인크. Prevention of disulfide bond reduction during recombinant production of polypeptides
WO2009012268A1 (en) 2007-07-16 2009-01-22 Genentech, Inc. Anti-cd79b antibodies and immunoconjugates and methods of use
TW200918089A (en) 2007-07-16 2009-05-01 Genentech Inc Humanized anti-CD79b antibodies and immunoconjugates and methods of use
CA2694488A1 (en) 2007-07-31 2009-02-05 Medimmune, Llc Multispecific epitope binding proteins and uses thereof
SI2769729T1 (en) 2007-09-04 2019-06-28 Compugen Ltd. Polypeptides and polynucleotides, and uses thereof as a drug target for producing drugs and biologics
KR101680906B1 (en) 2007-09-26 2016-11-30 추가이 세이야쿠 가부시키가이샤 Modified antibody constant region
AR068767A1 (en) 2007-10-12 2009-12-02 Novartis Ag ANTIBODIES AGAINST SCLEROSTIN, COMPOSITIONS AND METHODS OF USE OF THESE ANTIBODIES TO TREAT A PATHOLOGICAL DISORDER MEDIATIONED BY SCLEROSTIN
ES2565202T3 (en) 2007-10-16 2016-04-01 Zymogenetics, Inc. Combination of transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI) and anti-CD20 agents for the treatment of autoimmune diseases
JO3076B1 (en) * 2007-10-17 2017-03-15 Janssen Alzheimer Immunotherap Immunotherapy regimes dependent on apoe status
PL2567709T3 (en) 2007-11-02 2018-06-29 Novartis Ag Molecules and methods for modulating low-density-lipoprotein receptor-related protein 6 (LRP6)
KR101867606B1 (en) 2007-11-07 2018-06-18 제넨테크, 인크. Compositions and methods for treatment of microbial disorders
TWI580694B (en) 2007-11-30 2017-05-01 建南德克公司 Anti-vegf antibodies
US8795667B2 (en) * 2007-12-19 2014-08-05 Macrogenics, Inc. Compositions for the prevention and treatment of smallpox
EP3211010A1 (en) 2007-12-21 2017-08-30 Medimmune Limited Binding members for interleukin-4 receptor alpha (il-4r) - 173
US8092804B2 (en) 2007-12-21 2012-01-10 Medimmune Limited Binding members for interleukin-4 receptor alpha (IL-4Rα)-173
US7914785B2 (en) 2008-01-02 2011-03-29 Bergen Teknologieverforing As B-cell depleting agents, like anti-CD20 antibodies or fragments thereof for the treatment of chronic fatigue syndrome
EP2077281A1 (en) 2008-01-02 2009-07-08 Bergen Teknologioverforing AS Anti-CD20 antibodies or fragments thereof for the treatment of chronic fatigue syndrome
TWI472339B (en) 2008-01-30 2015-02-11 Genentech Inc Composition comprising antibody that binds to domain ii of her2 and acidic variants thereof
ES2587392T3 (en) 2008-01-31 2016-10-24 Genentech, Inc. Anti-CD79b and immunoconjugate antibodies and methods of use
AU2009211340B2 (en) * 2008-02-04 2013-09-12 Lay Line Genomics S.P.A. Anti-TrkA antibodies and derivatives thereof
MX2010008570A (en) 2008-02-05 2010-08-30 Bristol Myers Squibb Co Alpha 5 - beta 1 antibodies and their uses.
MX2010008578A (en) * 2008-02-08 2010-11-10 Medimmune Llc Anti-ifnar1 antibodies with reduced fc ligand affinity.
KR102070761B1 (en) * 2008-03-31 2020-01-29 제넨테크, 인크. Compositions and methods for treating and diagnosing asthma
MX2010010737A (en) 2008-04-02 2010-12-20 Macrogenics Inc Her2/neu-specific antibodies and methods of using same.
ES2589912T3 (en) 2008-04-02 2016-11-17 Macrogenics, Inc. Specific antibodies for the BCR complex and procedures for their use
KR20210113446A (en) 2008-05-02 2021-09-15 악셀레론 파마 인코포레이티드 Methods and compositions based on alk1 antagonists for modulating angiogenesis and pericyte coverage
US20110081347A1 (en) * 2008-06-04 2011-04-07 Macrogenics, Inc. Antibodies with Altered Binding to FcRn and Methods of Using Same
US9176310B2 (en) 2008-07-25 2015-11-03 Sloan Kettering Institute For Cancer Research Rapid confocal microscopy to support surgical procedures
BRPI0916668B1 (en) 2008-08-05 2021-12-28 Novartis Ag ISOLATED MONOCLONAL ANTIBODY, PHARMACEUTICAL COMPOSITION, ISOLATED NUCLEIC ACID AND VECTOR
AR072999A1 (en) 2008-08-11 2010-10-06 Medarex Inc HUMAN ANTIBODIES THAT JOIN GEN 3 OF LYMPHOCYTARY ACTIVATION (LAG-3) AND THE USES OF THESE
AU2009287165A1 (en) * 2008-08-29 2010-03-04 Symphogen A/S Method for cloning avian-derived antibodies
TW201438738A (en) 2008-09-16 2014-10-16 Genentech Inc Methods for treating progressive multiple sclerosis
EP2166021A1 (en) * 2008-09-16 2010-03-24 Ganymed Pharmaceuticals AG Monoclonal antibodies for treatment of cancer
AU2009294415B2 (en) 2008-09-19 2015-09-24 Medimmune Llc Antibodies directed to DLL4 and uses thereof
CN102245208B (en) 2008-10-14 2016-03-16 霍夫曼-拉罗奇有限公司 immunoglobulin variants and uses thereof
US9067981B1 (en) 2008-10-30 2015-06-30 Janssen Sciences Ireland Uc Hybrid amyloid-beta antibodies
US8298533B2 (en) 2008-11-07 2012-10-30 Medimmune Limited Antibodies to IL-1R1
CN102438652B (en) 2008-11-12 2014-08-13 米迪缪尼有限公司 Antibody formulation
WO2010057047A1 (en) * 2008-11-13 2010-05-20 Trubion Pharmaceutics, Inc. Cd37 immunotherapeutic combination therapies and uses thereof
HUE024872T2 (en) 2008-11-22 2016-02-29 Hoffmann La Roche Use of anti-vegf antibody in combination with chemotherapy for treating breast cancer
EP2373687B1 (en) 2008-12-03 2016-10-26 Genmab A/S Igg4 antibody variants having modifications in the constant region
CN102300874B (en) 2008-12-08 2015-08-05 卡姆普根有限公司 TMEM154 polypeptide and polynucleotide and as the purposes of drug targets producing medicine and biological products
PT4209510T (en) 2008-12-09 2024-04-02 Hoffmann La Roche Anti-pd-l1 antibodies and their use to enhance t-cell function
US8775090B2 (en) 2008-12-12 2014-07-08 Medimmune, Llc Crystals and structure of a human IgG Fc variant with enhanced FcRn binding
WO2010075249A2 (en) 2008-12-22 2010-07-01 Genentech, Inc. A method for treating rheumatoid arthritis with b-cell antagonists
MX2011006870A (en) 2008-12-23 2011-07-19 Genentech Inc Immunoglobulin variants with altered binding to protein a.
AU2009331528A1 (en) 2008-12-23 2011-08-11 Astrazeneca Ab Targeted binding agents directed to alpha5beta1 and uses thereof
JP2012514458A (en) 2008-12-31 2012-06-28 バイオジェン・アイデック・エムエイ・インコーポレイテッド Anti-lymphotoxin antibody
US20110142836A1 (en) * 2009-01-02 2011-06-16 Olav Mella B-cell depleting agents for the treatment of chronic fatigue syndrome
WO2010094499A1 (en) 2009-02-20 2010-08-26 Ganymed Pharmaceuticals Ag Methods and compositions for diagnosis and treatment of cancer
EP2398829A2 (en) * 2009-02-23 2011-12-28 Glenmark Pharmaceuticals S.A. Humanized antibodies that bind to cd19 and their uses
WO2010100200A2 (en) * 2009-03-05 2010-09-10 Novartis Ag Lyophilised antibody formulation
WO2010102175A1 (en) 2009-03-05 2010-09-10 Medarex, Inc. Fully human antibodies specific to cadm1
EA201101241A1 (en) 2009-03-06 2012-04-30 Калобайос Фармасьютиклз, Инк. TREATMENT OF LEUKEMIA AND CHRONIC MYELOPROLIFERATIVE DISEASES BY ERP3 Antibodies
AR075896A1 (en) 2009-03-20 2011-05-04 Genentech Inc ANTI-HER ANTIBODIES (EPIDERMAL GROWTH FACTOR)
MX345909B (en) 2009-03-25 2017-02-22 Genentech Inc Anti-fgfr3 antibodies and methods using same.
AU2010229479B2 (en) 2009-03-25 2013-03-28 Genentech, Inc. Novel anti-alpha5beta1 antibodies and uses thereof
TW201038286A (en) 2009-04-01 2010-11-01 Genentech Inc Anti-FcRH5 antibodies and immunoconjugates and methods of use
PT2417156E (en) 2009-04-07 2015-04-29 Roche Glycart Ag Trivalent, bispecific antibodies
US20100297127A1 (en) 2009-04-08 2010-11-25 Ghilardi Nico P Use of il-27 antagonists to treat lupus
NZ595792A (en) 2009-04-20 2014-01-31 Oxford Biotherapeutics Ltd Antibodies specific to cadherin-17
US9062116B2 (en) 2009-04-23 2015-06-23 Infinity Pharmaceuticals, Inc. Anti-fatty acid amide hydrolase-2 antibodies and uses thereof
US8715657B2 (en) 2009-04-27 2014-05-06 Novartis Ag Therapeutic antibodies binding IL12Rβ1
CN104725512A (en) 2009-04-27 2015-06-24 诺华股份有限公司 Compositions and methods for increasing muscle growth
CA2761438C (en) 2009-05-15 2017-12-12 University Health Network Compositions and methods for treating hematologic cancers targeting the sirp.alpha.-cd47 interaction
US9676845B2 (en) 2009-06-16 2017-06-13 Hoffmann-La Roche, Inc. Bispecific antigen binding proteins
JP6059985B2 (en) 2009-06-18 2017-01-11 ファイザー・インク Anti-NOTCH-1 antibody
CA2766403A1 (en) 2009-07-13 2011-01-20 Genentech, Inc. Diagnostic methods and compositions for treatment of cancer
AR077595A1 (en) 2009-07-27 2011-09-07 Genentech Inc COMBINATION TREATMENTS
EP3199551A3 (en) 2009-07-31 2017-10-18 E. R. Squibb & Sons, L.L.C. Fully human antibodies to btla
RU2567803C2 (en) 2009-07-31 2015-11-10 Дженентек, Инк. INHIBITION OF DISSEMINATION OF TUMOUR USING Bv8 OR G-CSF ANTAGONISTS
RU2017138926A (en) 2009-08-11 2019-02-11 Дженентек, Инк. PRODUCTION OF PROTEINS IN CULTURAL MEDIA WITHOUT GLUTAMINE
JP2013501526A (en) 2009-08-14 2013-01-17 ジェネンテック， インコーポレイテッド Biological markers for monitoring patient response to VEGF antagonists
SG178419A1 (en) 2009-08-15 2012-04-27 Genentech Inc Anti-angiogenesis therapy for the treatment of previously treated breast cancer
WO2011021146A1 (en) 2009-08-20 2011-02-24 Pfizer Inc. Osteopontin antibodies
EP3222287A1 (en) 2009-08-24 2017-09-27 Amunix Operating Inc. Coagulation factor ix compositions and methods of making and using same
JP5887270B2 (en) 2009-09-02 2016-03-16 ジェネンテック， インコーポレイテッド Mutant SMOOTHENED AND METHOD OF USING THE SAME
IN2012DN02485A (en) 2009-09-03 2015-08-28 Hoffmann La Roche
WO2011029823A1 (en) 2009-09-09 2011-03-17 Novartis Ag Monoclonal antibody reactive with cd63 when expressed at the surface of degranulated mast cells
MX2012002862A (en) 2009-09-11 2012-08-15 Genentech Inc Method to identify a patient with an increased likelihood of responding to an anti-cancer agent.
US20120302737A1 (en) 2009-09-16 2012-11-29 Genentech, Inc. Coiled coil and/or tether containing protein complexes and uses thereof
CA2772670A1 (en) 2009-09-17 2011-03-24 F. Hoffmann-La Roche Ag Methods and compositions for diagnostics use in cancer patients
US8926976B2 (en) 2009-09-25 2015-01-06 Xoma Technology Ltd. Modulators
CA2772945A1 (en) 2009-09-25 2011-03-31 Xoma Technology Ltd. Screening methods
US8568726B2 (en) 2009-10-06 2013-10-29 Medimmune Limited RSV specific binding molecule
TR201804897T4 (en) 2009-10-07 2018-06-21 Macrogenics Inc POLYPEPTIDES CONTAINING FC REGION WITH ADVANCED EFFECTOR FUNCTION DUE TO CHANGES OF FUCOSILATION SIZE AND METHODS FOR THEIR USE
EP2470569A1 (en) 2009-10-13 2012-07-04 Oxford Biotherapeutics Ltd. Antibodies against epha10
CA2777068C (en) 2009-10-14 2020-05-26 Kalobios Pharmaceuticals, Inc. Antibodies to epha3
WO2011050188A1 (en) 2009-10-22 2011-04-28 Genentech, Inc. Anti-hepsin antibodies and methods using same
WO2011056502A1 (en) 2009-10-26 2011-05-12 Genentech, Inc. Bone morphogenetic protein receptor type ii compositions and methods of use
WO2011056494A1 (en) 2009-10-26 2011-05-12 Genentech, Inc. Activin receptor-like kinase-1 antagonist and vegfr3 antagonist combinations
WO2011056497A1 (en) 2009-10-26 2011-05-12 Genentech, Inc. Activin receptor type iib compositions and methods of use
KR102624939B1 (en) 2009-11-02 2024-01-16 유니버시티 오브 워싱톤 스루 이츠 센터 포 커머셜리제이션 Therapeutic Nuclease Compositions and Methods
KR101898739B1 (en) 2009-11-04 2018-09-13 머크 샤프 앤드 돔 코포레이션 Engineered anti-tslp antibody
CA2780143A1 (en) 2009-11-05 2011-05-12 Genentech, Inc. Methods and composition for secretion of heterologous polypeptides
KR102317098B1 (en) 2009-11-11 2021-10-25 가니메드 파마슈티칼스 게엠베하 Antibodies specific for claudin 6 (cldn6)
EP2322555A1 (en) 2009-11-11 2011-05-18 Ganymed Pharmaceuticals AG Antibodies specific for claudin 6 (CLDN6)
MX359551B (en) 2009-11-24 2018-10-02 Medimmune Ltd Targeted binding agents against b7-h1.
EP2327725A1 (en) 2009-11-26 2011-06-01 InflaRx GmbH Anti-C5a binding moieties with high blocking activity
US9428586B2 (en) 2009-12-01 2016-08-30 Compugen Ltd Heparanase splice variant
BR112012013330A2 (en) 2009-12-02 2017-03-28 Acceleron Pharma Inc compositions and methods for increasing fc fusion protein serum half life
IN2012DN05175A (en) 2009-12-09 2015-10-23 Bayer Pharma AG
ES2565208T3 (en) 2009-12-11 2016-04-01 F. Hoffmann-La Roche Ag Anti-VEGF-C antibodies and methods of use thereof
EP2516458A1 (en) 2009-12-22 2012-10-31 Novartis AG Tetravalent cd47-antibody constant region fusion protein for use in therapy
MY161868A (en) 2009-12-23 2017-05-15 Genentech Inc Anti-bv8 antibodies and uses thereof
AU2011203879A1 (en) 2010-01-11 2012-08-02 Alexion Pharmaceuticals, Inc. Biomarkers of immunomodulatory effects in humans treated with anti-CD200 antibodies
EA201201120A1 (en) 2010-02-08 2013-03-29 Эдженсис, Инк. ANTIBODY AND MEDICINE CONJUGATES (ADC) THAT ARE ASSOCIATED WITH PROTEINS 161P2F10B
CN105001334A (en) 2010-02-10 2015-10-28 伊缪诺金公司 CD20 antibodies and uses thereof
WO2011098449A1 (en) 2010-02-10 2011-08-18 Novartis Ag Methods and compounds for muscle growth
CN102892779B (en) 2010-02-18 2016-12-21 基因泰克公司 Neuregulin antagonist and the purposes in treatment cancer thereof
KR20180028561A (en) 2010-02-23 2018-03-16 제넨테크, 인크. Anti-angiogenesis therapy for the treatment of ovarian cancer
US8802091B2 (en) 2010-03-04 2014-08-12 Macrogenics, Inc. Antibodies reactive with B7-H3 and uses thereof
EA030226B1 (en) 2010-03-04 2018-07-31 Макродженикс, Инк. Antibodies reactive with b7-h3, immunologically active fragments thereof and uses thereof
EP2371864A1 (en) 2010-03-23 2011-10-05 Ganymed Pharmaceuticals AG Monoclonal antibodies for treatment of cancer
TW201138823A (en) 2010-03-24 2011-11-16 Genentech Inc Anti-LRP6 antibodies
CA3079122A1 (en) 2010-03-26 2011-09-29 Trustees Of Dartmouth College Vista regulatory t cell mediator protein, vista binding agents and use thereof
US20150231215A1 (en) 2012-06-22 2015-08-20 Randolph J. Noelle VISTA Antagonist and Methods of Use
US10745467B2 (en) 2010-03-26 2020-08-18 The Trustees Of Dartmouth College VISTA-Ig for treatment of autoimmune, allergic and inflammatory disorders
AR080793A1 (en) 2010-03-26 2012-05-09 Roche Glycart Ag BISPECIFIC ANTIBODIES
CA2794745A1 (en) 2010-03-29 2011-10-06 Zymeworks, Inc. Antibodies with enhanced or suppressed effector function
WO2011133931A1 (en) 2010-04-22 2011-10-27 Genentech, Inc. Use of il-27 antagonists for treating inflammatory bowel disease
SG185383A1 (en) 2010-04-30 2012-12-28 Alexion Pharma Inc Anti-c5a antibodies and methods for using the antibodies
US9290573B2 (en) 2010-05-06 2016-03-22 Novartis Ag Therapeutic low density lipoprotein-related protein 6 (LRP6) multivalent antibodies
NZ603829A (en) 2010-05-06 2015-03-27 Novartis Ag Compositions and methods of use for therapeutic low density lipoprotein -related protein 6 (lrp6) antibodies
WO2011146568A1 (en) 2010-05-19 2011-11-24 Genentech, Inc. Predicting response to a her inhibitor
WO2011147834A1 (en) 2010-05-26 2011-12-01 Roche Glycart Ag Antibodies against cd19 and uses thereof
WO2011153243A2 (en) 2010-06-02 2011-12-08 Genentech, Inc. Anti-angiogenesis therapy for treating gastric cancer
WO2011153224A2 (en) 2010-06-02 2011-12-08 Genentech, Inc. Diagnostic methods and compositions for treatment of cancer
AU2011261362B2 (en) 2010-06-03 2016-06-09 Genentech, Inc. Immuno-pet imaging of antibodies and immunoconjugates and uses therefor
JP5940061B2 (en) 2010-06-18 2016-06-29 ジェネンテック， インコーポレイテッド Anti-AXL antibodies and methods of use
WO2011161119A1 (en) 2010-06-22 2011-12-29 F. Hoffmann-La Roche Ag Antibodies against insulin-like growth factor i receptor and uses thereof
WO2011161189A1 (en) 2010-06-24 2011-12-29 F. Hoffmann-La Roche Ag Anti-hepsin antibodies and methods of use
EP2404936A1 (en) 2010-07-06 2012-01-11 Ganymed Pharmaceuticals AG Cancer therapy using CLDN6 target-directed antibodies in vivo
MX2013000301A (en) 2010-07-09 2013-05-09 Biogen Idec Hemophilia Inc Chimeric clotting factors.
KR20130120439A (en) 2010-07-09 2013-11-04 제넨테크, 인크. Anti-neuropilin antibodies and methods of use
EP2409993A1 (en) 2010-07-19 2012-01-25 International-Drug-Development-Biotech Anti-CD19 antibody having ADCC function with improved glycosylation profile
EP2409712A1 (en) 2010-07-19 2012-01-25 International-Drug-Development-Biotech Anti-CD19 antibody having ADCC and CDC functions and improved glycosylation profile
WO2012010582A1 (en) 2010-07-21 2012-01-26 Roche Glycart Ag Anti-cxcr5 antibodies and methods of use
US20130177500A1 (en) 2010-07-23 2013-07-11 Trustee Of Boston University Anti-despr inhibitors as therapeutics for inhibition of pathological angiogenesis and tumor cell invasiveness and for molecular imaging and targeted delivery
CA2807127C (en) 2010-08-02 2019-02-12 Leslie S. Johnson Covalent diabodies and uses thereof
BR112013002535A2 (en) 2010-08-03 2019-09-24 Hoffmann La Roche biomarkers of chronic lymphocytic leukemia (cll)
BR112013002532A2 (en) 2010-08-05 2016-05-31 Hoffmann La Roche anti-mhc antibody anti-viral cytokine fusion protein
CA2808154A1 (en) 2010-08-13 2012-02-16 Medimmmune Limited Monomeric polypeptides comprising variant fc regions and methods of use
SG187746A1 (en) 2010-08-13 2013-03-28 Roche Glycart Ag Anti-fap antibodies and methods of use
CN103168049B (en) 2010-08-13 2015-10-07 罗切格利卡特公司 Anti-tenascin-C A2 antibody and using method
WO2012022734A2 (en) 2010-08-16 2012-02-23 Medimmune Limited Anti-icam-1 antibodies and methods of use
DK2606070T3 (en) 2010-08-20 2017-03-27 Novartis Ag Antibodies for the epidermal growth factor receptor 3 (HER3)
EP2609111B1 (en) 2010-08-24 2017-11-01 F. Hoffmann-La Roche AG Bispecific antibodies comprising a disulfide stabilized-fv fragment
ES2553262T3 (en) 2010-08-25 2015-12-07 F. Hoffmann-La Roche Ag Antibodies against IL-18R1 and their uses
KR20130103734A (en) 2010-08-31 2013-09-24 제넨테크, 인크. Biomarkers and methods of treatment
WO2012028683A1 (en) 2010-09-02 2012-03-08 Novartis Ag Antibody gel system for sustained drug delivery
WO2012034039A2 (en) 2010-09-10 2012-03-15 Apexigen, Inc. Anti-il-1 beta antibodies and methods of use
WO2012035518A1 (en) 2010-09-17 2012-03-22 Compugen Ltd. Compositions and methods for treatment of drug resistant multiple myeloma
KR20130130709A (en) 2010-09-29 2013-12-02 어젠시스 인코포레이티드 Antibody drug conjugates (adc) that bind to 191p4d12 proteins
JP2013543384A (en) 2010-10-05 2013-12-05 ノバルティス アーゲー Anti-IL12Rbeta1 antibody and its use in the treatment of autoimmune and inflammatory diseases
DK2625197T3 (en) 2010-10-05 2016-10-03 Genentech Inc Smoothened MUTANT AND METHODS OF USING THE SAME
EP2635607B1 (en) 2010-11-05 2019-09-04 Zymeworks Inc. Stable heterodimeric antibody design with mutations in the fc domain
TW201300417A (en) 2010-11-10 2013-01-01 Genentech Inc Methods and compositions for neural disease immunotherapy
WO2012075581A1 (en) 2010-12-06 2012-06-14 Ym Biosciences Inc. Antibodies selective for cells presenting erbb2 at high density
BR112013015122A2 (en) 2010-12-15 2017-05-30 Wyeth Llc anti-notch1 antibodies
KR101615420B1 (en) 2010-12-16 2016-04-26 제넨테크, 인크. Diagnosis and treatments relating to th2 inhibition
EA201790664A1 (en) 2010-12-20 2017-07-31 Дженентек, Инк. ANTIBODIES AGAINST MEZOTELINE AND IMMUNOCONJUGATES
MX2013007168A (en) 2010-12-22 2013-11-04 Genentech Inc Anti-pcsk9 antibodies and methods of use.
JP5766296B2 (en) 2010-12-23 2015-08-19 エフ．ホフマン−ラ ロシュ アーゲーＦ． Ｈｏｆｆｍａｎｎ−Ｌａ Ｒｏｃｈｅ Ａｋｔｉｅｎｇｅｓｅｌｌｓｃｈａｆｔ Polypeptide-polynucleotide complexes and their use in targeted delivery of effector components
WO2012092539A2 (en) 2010-12-31 2012-07-05 Takeda Pharmaceutical Company Limited Antibodies to dll4 and uses thereof
EP2668209B8 (en) 2011-01-24 2021-08-11 Gilead Sciences, Inc. Antibodies selective for cells presenting egfr at high density
US9447187B2 (en) 2011-02-03 2016-09-20 Alexion Pharmaceuticals, Inc. Use of an anti-CD200 antibody for prolonging the survival of allografts
MX353143B (en) 2011-02-28 2017-12-20 Genentech Inc Biological markers and methods for predicting response to b-cell antagonists.
CN103403025B (en) 2011-02-28 2016-10-12 弗·哈夫曼-拉罗切有限公司 Monovalent antigen binding protein
ES2549638T3 (en) 2011-02-28 2015-10-30 F. Hoffmann-La Roche Ag Antigen binding proteins
EP2681243B1 (en) 2011-03-03 2018-09-05 Apexigen, Inc. Anti-il-6 receptor antibodies and methods of use
US8609818B2 (en) 2011-03-10 2013-12-17 Omeros Corporation Generation of anti-FN14 monoclonal antibodies by ex-vivo accelerated antibody evolution
JP2014516511A (en) 2011-04-07 2014-07-17 ジェネンテック， インコーポレイテッド Anti-FGFR4 antibody and method of use
PL2697257T3 (en) 2011-04-13 2017-04-28 Bristol-Myers Squibb Company Fc fusion proteins comprising novel linkers or arrangements
CN105601741A (en) 2011-04-15 2016-05-25 卡姆普根有限公司 Polypeptides and polynucleotides, and uses thereof for treatment of immune related disorders and cancer
AU2012245439B2 (en) 2011-04-20 2017-04-06 Acceleron Pharma, Inc. Endoglin polypeptides and uses thereof
KR101970025B1 (en) 2011-04-20 2019-04-17 메디뮨 엘엘씨 Antibodies and other molecules that bind b7-h1 and pd-1
WO2012146630A1 (en) 2011-04-29 2012-11-01 F. Hoffmann-La Roche Ag N-terminal acylated polypeptides, methods for their production and uses thereof
ES2709654T3 (en) 2011-04-29 2019-04-17 Apexigen Inc Anti-CD40 antibodies and methods of use
AU2012249360B2 (en) 2011-04-29 2015-12-24 University Of Washington Therapeutic nuclease compositions and methods
WO2012155019A1 (en) 2011-05-12 2012-11-15 Genentech, Inc. Multiple reaction monitoring lc-ms/ms method to detect therapeutic antibodies in animal samples using framework signature pepides
PL2707390T3 (en) 2011-05-13 2016-06-30 Ganymed Pharmaceuticals Ag Antibodies for treatment of cancer expressing claudin 6
WO2012156309A1 (en) 2011-05-13 2012-11-22 Millegen Antibodies against her3
MX339662B (en) 2011-05-16 2016-06-03 Genentech Inc Fgfr1 agonists and methods of use.
EP2710032B1 (en) 2011-05-20 2019-12-25 Momenta Pharmaceuticals, Inc. Modified glycoproteins
CA2836873C (en) 2011-05-21 2019-10-22 Macrogenics, Inc. Deimmunized serum-binding domains and their use for extending serum half-life
SG195072A1 (en) 2011-05-21 2013-12-30 Macrogenics Inc Cd3-binding molecules capable of binding to human and non-human cd3
CN106432506A (en) 2011-05-24 2017-02-22 泽恩格尼亚股份有限公司 Multivalent and monovalent multispecific complexes and their uses
WO2012167143A1 (en) 2011-06-03 2012-12-06 Xoma Technology Ltd. Antibodies specific for tgf-beta
WO2012170740A2 (en) 2011-06-07 2012-12-13 University Of Hawaii Biomarker of asbestos exposure and mesothelioma
WO2012170742A2 (en) 2011-06-07 2012-12-13 University Of Hawaii Treatment and prevention of cancer with hmgb1 antagonists
EP3527218A1 (en) 2011-06-10 2019-08-21 Bioverativ Therapeutics Inc. Pro-coagulant compounds and methods of use thereof
WO2012172495A1 (en) 2011-06-14 2012-12-20 Novartis Ag Compositions and methods for antibodies targeting tem8
CN103596984B (en) 2011-06-15 2016-04-13 霍夫曼-拉罗奇有限公司 The antibody of Anti-human EPO receptor and using method
KR20140030250A (en) 2011-06-16 2014-03-11 노파르티스 아게 Soluble proteins for use as therapeutics
EP2723376B1 (en) 2011-06-22 2018-12-05 INSERM (Institut National de la Santé et de la Recherche Médicale) Anti-axl antibodies and uses thereof
PE20141212A1 (en) 2011-06-22 2014-09-19 Hoffmann La Roche ELIMINATION OF TARGET CELLS BY VIRUS SPECIFIC CYTOTOXIC T CELLS USING COMPLEXES INCLUDING CLASS 1 MHC
CA2839480A1 (en) 2011-06-22 2012-12-27 Inserm (Institut National De La Sante Et De La Recherche Medicale) Anti-axl antibodies and uses thereof
CN103649121B (en) 2011-06-28 2016-10-19 牛津生物疗法有限公司 Antibody for ADP-ribosyl cyclase 2
SG194932A1 (en) 2011-06-30 2013-12-30 Genentech Inc Anti-c-met antibody formulations
WO2013001517A1 (en) 2011-06-30 2013-01-03 Compugen Ltd. Polypeptides and uses thereof for treatment of autoimmune disorders and infection
ES2692519T3 (en) 2011-07-01 2018-12-04 Novartis Ag Method to treat metabolic disorders
LT2731677T (en) 2011-07-11 2018-07-10 Glenmark Pharmaceuticals S.A. Antibodies that bind to ox40 and their uses
US9738707B2 (en) 2011-07-15 2017-08-22 Biogen Ma Inc. Heterodimeric Fc regions, binding molecules comprising same, and methods relating thereto
WO2013010955A1 (en) 2011-07-15 2013-01-24 Morphosys Ag Antibodies that are cross-reactive for macrophage migration inhibitory factor (mif) and d-dopachrome tautomerase (d-dt)
US20130022551A1 (en) 2011-07-22 2013-01-24 Trustees Of Boston University DEspR ANTAGONISTS AND AGONISTS AS THERAPEUTICS
EP3783028A1 (en) 2011-08-12 2021-02-24 Omeros Corporation Anti-fzd10 monoclonal antibodies and methods for their use
EP2756094B1 (en) 2011-08-15 2017-12-27 Medlmmune, LLC Anti-b7-h4 antibodies and their uses
KR20140068877A (en) 2011-08-17 2014-06-09 제넨테크, 인크. Inhibition of angiogenesis in refractory tumors
EP2744824A1 (en) 2011-08-17 2014-06-25 F.Hoffmann-La Roche Ag Neuregulin antibodies and uses thereof
CN103890006A (en) 2011-08-23 2014-06-25 罗切格利卡特公司 Anti-mcsp antibodies
GB201212550D0 (en) 2012-07-13 2012-08-29 Novartis Ag B cell assay
UY34317A (en) 2011-09-12 2013-02-28 Genzyme Corp T cell antireceptor antibody (alpha) / ß
WO2013039954A1 (en) 2011-09-14 2013-03-21 Sanofi Anti-gitr antibodies
EP2756300A1 (en) 2011-09-15 2014-07-23 F.Hoffmann-La Roche Ag Methods of promoting differentiation
CA2846630A1 (en) 2011-09-19 2013-03-28 Genentech, Inc. Combination treatments comprising c-met antagonists and b-raf antagonists
RS55822B1 (en) 2011-09-30 2017-08-31 Dana-Farber Cancer Institute Inc Therapeutic peptides
CA2849011A1 (en) 2011-10-05 2013-04-11 Genentech, Inc. Methods of treating liver conditions using notch2 antagonists
EP3653222A1 (en) 2011-10-14 2020-05-20 Novartis AG Antibodies and methods for wnt pathway-related diseases
PL2766393T3 (en) 2011-10-14 2018-11-30 F.Hoffmann-La Roche Ag ANTI-HtrA1 ANTIBODIES AND METHODS OF USE
RU2014119426A (en) 2011-10-15 2015-11-20 Дженентек, Инк. WAYS OF APPLICATION OF SCD1 ANTAGONISTS
WO2013059531A1 (en) 2011-10-20 2013-04-25 Genentech, Inc. Anti-gcgr antibodies and uses thereof
KR20140097205A (en) 2011-10-28 2014-08-06 제넨테크, 인크. Therapeutic combinations and methods of treating melanoma
EP2773373B1 (en) 2011-11-01 2018-08-22 Bionomics, Inc. Methods of blocking cancer stem cell growth
CA2853951A1 (en) 2011-11-01 2013-05-10 Bionomics, Inc. Antibodies and methods of treating cancer
AU2012332593B2 (en) 2011-11-01 2016-11-17 Bionomics, Inc. Anti-GPR49 antibodies
WO2013067057A1 (en) 2011-11-01 2013-05-10 Bionomics, Inc. Anti-gpr49 antibodies
KR102027603B1 (en) 2011-11-02 2019-10-01 아펙시젠, 인코포레이티드 Anti-kdr antibodies and methods of use
RU2675319C2 (en) 2011-11-04 2018-12-18 Займворкс Инк. STABLE HETERODIMERIC ANTIBODY DESIGN WITH MUTATIONS IN Fc DOMAIN
CN103906533A (en) 2011-11-07 2014-07-02 米迪缪尼有限公司 Multispecific and multivalent binding proteins and uses thereof
JP2014533700A (en) 2011-11-21 2014-12-15 ジェネンテック， インコーポレイテッド Purification of anti-c-MET antibody
CN104302669A (en) 2011-11-23 2015-01-21 伊格尼卡生物治疗公司 Anti-CD98 antibodies and methods of use thereof
AR088941A1 (en) 2011-11-23 2014-07-16 Bayer Ip Gmbh ANTI-FGFR2 ANTIBODIES AND THEIR USES
EP3712173B1 (en) 2011-12-05 2023-07-12 X-Body, Inc. Pdgf receptor beta binding polypeptides
EA201491107A1 (en) 2011-12-05 2014-11-28 Новартис Аг ANTIBODIES TO THE RECEPTOR EPIDERMAL GROWTH FACTOR 3 (HER3), DIRECTED TO DOMAIN II HER3
UY34487A (en) 2011-12-05 2013-07-31 Novartis Ag ANTIBODIES FOR EPIDERMAL GROWTH FACTOR RECEIVER 3 (HER3)
WO2013083497A1 (en) 2011-12-06 2013-06-13 F. Hoffmann-La Roche Ag Antibody formulation
RU2685867C2 (en) 2011-12-15 2019-04-23 Алтернатив Инновейтив Текнолоджиз Ллц Hybrid proteins and protein conjugates based on heat shock protein-70 (hsp70) and methods for use thereof (versions)
WO2013096291A2 (en) 2011-12-20 2013-06-27 Medimmune, Llc Modified polypeptides for bispecific antibody scaffolds
MX2014007665A (en) 2011-12-21 2015-03-05 Novartis Ag Compositions and methods for antibodies targeting factor p.
AR089434A1 (en) 2011-12-23 2014-08-20 Genentech Inc PROCEDURE TO PREPARE FORMULATIONS WITH HIGH CONCENTRATION OF PROTEINS
WO2013101771A2 (en) 2011-12-30 2013-07-04 Genentech, Inc. Compositions and method for treating autoimmune diseases
EP3663314A1 (en) 2012-01-09 2020-06-10 The Scripps Research Institute Humanized antibodies with ultralong cdr3s
CN104520321A (en) 2012-01-09 2015-04-15 斯克利普斯研究所 Ultralong complementarity determining regions and uses thereof
RS59670B1 (en) 2012-01-12 2020-01-31 Bioverativ Therapeutics Inc Chimeric factor viii polypeptides and uses thereof
WO2013106765A1 (en) 2012-01-13 2013-07-18 Genentech, Inc. Biological markers for identifying patients for treatment with vegf antagonists
MX2014008699A (en) 2012-01-18 2014-11-21 Genentech Inc Methods of using fgf19 modulators.
CA2862422A1 (en) 2012-01-18 2013-07-25 Genentech, Inc. Anti-lrp5 antibodies and methods of use
CA2863066A1 (en) 2012-01-31 2013-08-08 Genentech, Inc. Anti-ig-e m1' antibodies and methods using same
WO2013114367A2 (en) 2012-02-01 2013-08-08 Compugen Ltd. C10rf32 antibodies, and uses thereof for treatment of cancer
BR112014019579A2 (en) 2012-02-10 2019-10-15 Genentech, Inc SINGLE CHAIN ANTIBODY, POLYNUCLEOTIDE, VECTOR, HOST CELL, METHOD OF PRODUCTION OF A SINGLE CHAIN ANTIBODY, HETEROMULTYMER AND METHOD OF PRODUCTION
CN104718220A (en) 2012-02-11 2015-06-17 霍夫曼-拉罗奇有限公司 R-spondin translocations and methods using the same
MY188897A (en) 2012-02-15 2022-01-12 Bioverativ Therapeutics Inc Factor viii compositions and methods of making and using same
EP3549953A1 (en) 2012-02-15 2019-10-09 Bioverativ Therapeutics Inc. Recombinant factor viii proteins
EP2814587B1 (en) 2012-02-15 2018-05-02 F.Hoffmann-La Roche Ag Fc-receptor based affinity chromatography
EP2831115A1 (en) 2012-03-27 2015-02-04 F. Hoffmann-La Roche AG Diagnosis and treatments relating to her3 inhibitors
RS57413B1 (en) 2012-03-28 2018-09-28 Sanofi Sa Antibodies to bradykinin b1 receptor ligands
AR090549A1 (en) 2012-03-30 2014-11-19 Genentech Inc ANTI-LGR5 AND IMMUNOCATE PLAYERS
MX2014011582A (en) 2012-03-30 2014-11-21 Genentech Inc Diagnostic methods and compositions for treatment of cancer.
EP2834270B1 (en) 2012-04-05 2019-10-30 AC Immune S.A. Humanized tau antibody
EP2841452B1 (en) 2012-04-25 2023-04-12 Momenta Pharmaceuticals, Inc. Modified glycoproteins
AR090903A1 (en) 2012-05-01 2014-12-17 Genentech Inc ANTI-PMEL ANTIBODIES AND IMMUNOCADES17
EP2847216A1 (en) 2012-05-07 2015-03-18 Sanofi Methods for preventing biofilm formation
WO2013167153A1 (en) 2012-05-09 2013-11-14 Ganymed Pharmaceuticals Ag Antibodies useful in cancer diagnosis
WO2013170191A1 (en) 2012-05-11 2013-11-14 Genentech, Inc. Methods of using antagonists of nad biosynthesis from nicotinamide
JP6294311B2 (en) 2012-05-23 2018-03-14 ジェネンテック， インコーポレイテッド How to select a treatment
WO2013175276A1 (en) 2012-05-23 2013-11-28 Argen-X B.V Il-6 binding molecules
EP2855520B1 (en) 2012-06-04 2018-09-26 Novartis AG Site-specific labeling methods and molecules produced thereby
CA2875246A1 (en) 2012-06-08 2013-12-12 Biogen Idec Ma Inc. Procoagulant compounds
EA201492163A1 (en) 2012-06-08 2015-08-31 Гленмарк Фармасьютикалс С.А. HUMANIZED ANTIBODIES TO TRKA, CONTAINING AMINO ACID SUBSTITUTES
WO2013185114A2 (en) 2012-06-08 2013-12-12 Biogen Idec Ma Inc. Chimeric clotting factors
CN104364266A (en) 2012-06-15 2015-02-18 霍夫曼-拉罗奇有限公司 Anti-PCSK9 antibodies, formulations, dosing, and methods of use
EP2863954A1 (en) 2012-06-21 2015-04-29 Indiana University Research and Technology Corporation Incretin receptor ligand polypeptide fc-region fusion polypeptides and conjugates with altered fc-effector function
US9890215B2 (en) 2012-06-22 2018-02-13 King's College London Vista modulators for diagnosis and treatment of cancer
WO2014004586A1 (en) 2012-06-25 2014-01-03 Zymeworks Inc. Process and methods for efficient manufacturing of highly pure asymmetric antibodies in mammalian cells
EP2867253B1 (en) 2012-06-27 2016-09-14 F. Hoffmann-La Roche AG Method for selection and production of tailor-made highly selective and multi-specific targeting entities containing at least two different binding entities and uses thereof
JP6309002B2 (en) 2012-06-27 2018-04-11 エフ．ホフマン−ラ ロシュ アーゲーＦ． Ｈｏｆｆｍａｎｎ−Ｌａ Ｒｏｃｈｅ Ａｋｔｉｅｎｇｅｓｅｌｌｓｃｈａｆｔ Methods and uses for making antibody Fc region conjugates comprising at least one binding entity that specifically binds to a target
AR091649A1 (en) 2012-07-02 2015-02-18 Bristol Myers Squibb Co OPTIMIZATION OF ANTIBODIES THAT FIX THE LYMPHOCYTE ACTIVATION GEN 3 (LAG-3) AND ITS USES
HUE030858T2 (en) 2012-07-04 2017-06-28 Hoffmann La Roche Covalently linked antigen-antibody conjugates
RU2630664C2 (en) 2012-07-04 2017-09-11 Ф. Хоффманн-Ля Рош Аг Theophylline antibodies and methods for their application
CA2872192A1 (en) 2012-07-04 2014-01-09 F. Hoffmann-La Roche Ag Anti-biotin antibodies and methods of use
CN104428416B (en) 2012-07-05 2019-01-29 弗·哈夫曼-拉罗切有限公司 Expression and excretory system
WO2014008480A2 (en) 2012-07-06 2014-01-09 Biogen Idec Ma Inc. Cell line expressing single chain factor viii polypeptides and uses thereof
EA201590171A1 (en) 2012-07-09 2015-09-30 Дженентек, Инк. IMMUNOCONGATES CONTAINING ANTIBODIES TO CD79b
JP6297550B2 (en) 2012-07-09 2018-03-20 ジェネンテック， インコーポレイテッド Immune complex comprising anti-CD79B antibody
PE20150615A1 (en) 2012-07-09 2015-05-28 Genentech Inc IMMUNOCONJUGATES INCLUDING AN ANTI-CD22 ANTIBODY LINKED TO A PYRROLOBENZODIAZEPINE
AU2013288931A1 (en) 2012-07-09 2014-12-11 Genentech, Inc. Immunoconjugates comprising anti-CD22 antibodies
SG10201913893XA (en) 2012-07-11 2020-03-30 Bioverativ Therapeutics Inc Factor viii complex with xten and von willebrand factor protein, and uses thereof
EP3381943B1 (en) 2012-07-25 2022-03-16 Celldex Therapeutics, Inc. Anti-kit antibodies and uses thereof
JP6307077B2 (en) 2012-08-02 2018-04-04 エフ．ホフマン−ラ ロシュ アーゲーＦ． Ｈｏｆｆｍａｎｎ−Ｌａ Ｒｏｃｈｅ Ａｋｔｉｅｎｇｅｓｅｌｌｓｃｈａｆｔ Method for producing soluble FcR as an Fc fusion with an inactive immunoglobulin Fc region and use thereof
SG10201701424QA (en) 2012-08-23 2017-04-27 Agensys Inc Antibody drug conjugates (adc) that bind to 158p1d7 proteins
EP2892558B1 (en) 2012-09-07 2019-04-10 The Trustees Of Dartmouth College Vista modulators for diagnosis and treatment of cancer
JOP20200308A1 (en) 2012-09-07 2017-06-16 Novartis Ag IL-18 binding molecules
US9790268B2 (en) 2012-09-12 2017-10-17 Genzyme Corporation Fc containing polypeptides with altered glycosylation and reduced effector function
RS64317B1 (en) 2012-09-12 2023-08-31 Genzyme Corp Fc containing polypeptides with altered glycosylation and reduced effector function
WO2014059028A1 (en) 2012-10-09 2014-04-17 Igenica, Inc. Anti-c16orf54 antibodies and methods of use thereof
WO2014070934A1 (en) 2012-10-30 2014-05-08 Apexigen, Inc. Anti-cd40 antibodies and methods of use
EP2914621B1 (en) 2012-11-05 2023-06-07 Foundation Medicine, Inc. Novel ntrk1 fusion molecules and uses thereof
CA2884431A1 (en) 2012-11-08 2014-05-15 F. Hoffmann-La Roche Ag Her3 antigen binding proteins binding to the beta-hairpin of her3
MA38176A1 (en) 2012-11-13 2017-06-30 Genentech Inc Novel anti-haemagglutinin antibody, useful for the treatment, inhibition or prevention of viral influenza infection
EP2733153A1 (en) 2012-11-15 2014-05-21 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for the preparation of immunoconjugates and uses thereof
US9914785B2 (en) 2012-11-28 2018-03-13 Zymeworks Inc. Engineered immunoglobulin heavy chain-light chain pairs and uses thereof
WO2014084859A1 (en) 2012-11-30 2014-06-05 Novartis Ag Molecules and methods for modulating tmem16a activities
EP3851454A1 (en) 2012-12-05 2021-07-21 Novartis AG Compositions and methods for antibodies targeting epo
PT2928923T (en) 2012-12-10 2020-03-27 Biogen Ma Inc Anti-blood dendritic cell antigen 2 antibodies and uses thereof
EP2934576A1 (en) 2012-12-19 2015-10-28 The Johns Hopkins University Anti-human b7-h4 antibodies and their uses
CA2889788A1 (en) 2012-12-21 2014-06-26 F. Hoffmann-La Roche Ag Disulfide-linked multivalent mhc class i comprising multi-function proteins
KR20150100715A (en) 2012-12-21 2015-09-02 앰플리뮨, 인크. Anti-h7cr antibodies
CA2896894A1 (en) 2013-01-02 2014-07-10 Glenmark Pharmaceuticals S.A. Antibodies that bind to tl1a and their uses
CA2898326C (en) 2013-01-18 2022-05-17 Foundation Medicine, Inc. Methods of treating cholangiocarcinoma
WO2014116749A1 (en) 2013-01-23 2014-07-31 Genentech, Inc. Anti-hcv antibodies and methods of using thereof
US10568975B2 (en) 2013-02-05 2020-02-25 The Johns Hopkins University Nanoparticles for magnetic resonance imaging tracking and methods of making and using thereof
EP3620468A1 (en) 2013-02-05 2020-03-11 EngMab Sàrl Method for the selection of antibodies against bcma
EP2762496A1 (en) 2013-02-05 2014-08-06 EngMab AG Method for the selection of antibodies against BCMA
WO2014124258A2 (en) 2013-02-08 2014-08-14 Irm Llc Specific sites for modifying antibodies to make immunoconjugates
MX2015010146A (en) 2013-02-08 2016-05-31 Novartis Ag Specific sites for modifying antibodies to make immunoconjugates.
SG11201505330QA (en) 2013-02-08 2015-08-28 Novartis Ag Anti-il-17a antibodies and their use in treating autoimmune and inflammatory disorders
ES2870802T3 (en) 2013-02-12 2021-10-27 Bristol Myers Squibb Co High pH Protein Refolding Methods
US10183967B2 (en) 2013-02-12 2019-01-22 Bristol-Myers Squibb Company Tangential flow filtration based protein refolding methods
SI3889173T1 (en) 2013-02-15 2023-11-30 Bioverativ Therapeutics Inc. Optimized factor viii gene
WO2014128235A1 (en) 2013-02-22 2014-08-28 F. Hoffmann-La Roche Ag Methods of treating cancer and preventing drug resistance
AR094873A1 (en) 2013-02-25 2015-09-02 Genentech Inc METHODS AND COMPOSITIONS TO DETECT AND TREAT MUTANTS OF AKT RESISTANT TO DRUGS
KR20150123811A (en) 2013-02-26 2015-11-04 로슈 글리카트 아게 Anti-mcsp antibodies
US9487587B2 (en) 2013-03-05 2016-11-08 Macrogenics, Inc. Bispecific molecules that are immunoreactive with immune effector cells of a companion animal that express an activating receptor and cells that express B7-H3 and uses thereof
MX2015011428A (en) 2013-03-06 2016-02-03 Genentech Inc Methods of treating and preventing cancer drug resistance.
KR20240023184A (en) 2013-03-11 2024-02-20 젠자임 코포레이션 Hyperglycosylated binding polypeptides
US9498532B2 (en) 2013-03-13 2016-11-22 Novartis Ag Antibody drug conjugates
US9562099B2 (en) 2013-03-14 2017-02-07 Genentech, Inc. Anti-B7-H4 antibodies and immunoconjugates
HUE055845T2 (en) 2013-03-14 2021-12-28 Macrogenics Inc Bispecific molecules that are immunoreactive with immune effector cells that express an activating receptor
WO2014153030A2 (en) 2013-03-14 2014-09-25 Genentech, Inc. Methods of treating cancer and preventing cancer drug resistance
CA2902865A1 (en) 2013-03-14 2014-10-02 Genentech, Inc. Anti-b7-h4 antibodies and immunoconjugates
EP3611189A1 (en) 2013-03-14 2020-02-19 Novartis AG Antibodies against notch 3
MX2015010854A (en) 2013-03-14 2016-07-20 Genentech Inc Combinations of a mek inhibitor compound with an her3/egfr inhibitor compound and methods of use.
AR095517A1 (en) 2013-03-15 2015-10-21 Genentech Inc ANTIBODIES AGAINST THE CHEMIOATRAYENT RECEIVER EXPRESSED IN T HELPER 2 CELLS (ANTI-CRTh2) AND METHODS OF USE
EA201890895A1 (en) 2013-03-15 2019-02-28 Зинджения, Инк. MULTIVALENT AND MONOVALENT MULTIS-SPECIFIC COMPLEXES AND THEIR APPLICATION
JP6456356B2 (en) 2013-03-15 2019-01-23 ジェネンテック， インコーポレイテッド IL-22 polypeptides and IL-22 Fc fusion proteins and methods of use
NZ629816A (en) 2013-03-15 2017-07-28 Dana Farber Cancer Inst Inc Therapeutic peptides
AU2014228938B2 (en) 2013-03-15 2019-05-02 Bioverativ Therapeutics Inc. Factor IX polypeptide formulations
EP2968537A1 (en) 2013-03-15 2016-01-20 Genentech, Inc. Methods of treating cancer and preventing cancer drug resistance
EP3514178A1 (en) 2013-03-15 2019-07-24 Novartis AG Antibody drug conjugates
US10035859B2 (en) 2013-03-15 2018-07-31 Biogen Ma Inc. Anti-alpha V beta 6 antibodies and uses thereof
PE20151750A1 (en) 2013-03-15 2015-12-07 Genentech Inc COMPOSITIONS AND METHODS FOR THE DIAGNOSIS AND TREATMENT OF HEPATIC CANCER
WO2014150877A2 (en) 2013-03-15 2014-09-25 Ac Immune S.A. Anti-tau antibodies and methods of use
US20160017041A1 (en) 2013-03-15 2016-01-21 Biogen Ma Inc. Treatment and prevention of acute kidney injury using anti-alpha v beta 5 antibodies
US10035860B2 (en) 2013-03-15 2018-07-31 Biogen Ma Inc. Anti-alpha V beta 6 antibodies and uses thereof
KR102158924B1 (en) 2013-03-15 2020-09-22 제넨테크, 인크. Biomarkers and methods of treating pd-1 and pd-l1 related conditions
CA2908653A1 (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
CA2904806C (en) 2013-04-29 2021-11-23 F. Hoffmann-La Roche Ag Human fcrn-binding modified antibodies and methods of use
ES2871383T3 (en) 2013-04-29 2021-10-28 Hoffmann La Roche Modified Asymmetric Antibodies Binding to the Fc Receptor and Procedures for Use
US9708406B2 (en) 2013-05-20 2017-07-18 Genentech, Inc. Anti-transferrin receptor antibodies and methods of use
BR112015029395A2 (en) 2013-05-24 2017-09-19 Medimmune Llc ANTI-B7-H5 ANTIBODIES AND THEIR USES
US10053510B2 (en) 2013-05-24 2018-08-21 Promis Neurosciences Inc. FasR antibodies and methods of use
US11098105B2 (en) 2013-05-31 2021-08-24 Zymeworks Inc. Heteromultimers with reduced or silenced effector function
PE20160716A1 (en) 2013-06-14 2016-08-11 Bayer Pharma AG ANTI-TWEAKR ANTIBODY AND ITS USES
AR096601A1 (en) 2013-06-21 2016-01-20 Novartis Ag ANTIBODIES OF LEXINED OXIDATED LDL RECEIVER 1 AND METHODS OF USE
US9562101B2 (en) 2013-06-21 2017-02-07 Novartis Ag Lectin-like oxidized LDL receptor 1 antibodies and methods of use
CN105814074B (en) 2013-07-18 2020-04-21 图鲁斯生物科学有限责任公司 Humanized antibodies with ultralong complementarity determining regions
WO2015017146A2 (en) 2013-07-18 2015-02-05 Fabrus, Inc. Antibodies with ultralong complementarity determining regions
WO2015014376A1 (en) 2013-07-31 2015-02-05 Biontech Ag Diagnosis and therapy of cancer involving cancer stem cells
EP3027220A1 (en) 2013-08-01 2016-06-08 Agensys, Inc. Antibody drug conjugates (adc) that bind to cd37 proteins
PT3027651T (en) 2013-08-01 2019-05-31 Five Prime Therapeutics Inc Afucosylated anti-fgfr2iiib antibodies
EP3875106A1 (en) 2013-08-08 2021-09-08 Bioverativ Therapeutics Inc. Purification of chimeric fviii molecules
UA116479C2 (en) 2013-08-09 2018-03-26 Макродженікс, Інк. Bi-specific monovalent fc diabodies that are capable of binding cd32b and cd79b and uses thereof
US11384149B2 (en) 2013-08-09 2022-07-12 Macrogenics, Inc. Bi-specific monovalent Fc diabodies that are capable of binding CD32B and CD79b and uses thereof
KR20160042438A (en) 2013-08-12 2016-04-19 제넨테크, 인크. Compositions and method for treating complement-associated conditions
JP6696899B2 (en) 2013-08-13 2020-05-20 サノフイＳａｎｏｆｉ Antibody against plasminogen activator inhibitor-1 (PAI-1) and use thereof
TW201722994A (en) 2013-08-13 2017-07-01 賽諾菲公司 Antibodies to Plasminogen Activator Inhibitor-1 (PAI-1) and uses thereof
WO2015023891A2 (en) 2013-08-14 2015-02-19 Biogen Idec Ma Inc. Factor viii-xten fusions and uses thereof
SG10201801063TA (en) 2013-08-14 2018-04-27 Novartis Ag Methods of treating sporadic inclusion body myositis
EP3036320B2 (en) 2013-08-19 2024-04-03 Biogen MA Inc. Control of protein glycosylation by culture medium supplementation and cell culture process parameters
EP3036262A4 (en) 2013-08-22 2017-03-01 Acceleron Pharma Inc. Tgf-beta receptor type ii variants and uses thereof
EP2839842A1 (en) 2013-08-23 2015-02-25 MacroGenics, Inc. Bi-specific monovalent diabodies that are capable of binding CD123 and CD3 and uses thereof
EP2840091A1 (en) 2013-08-23 2015-02-25 MacroGenics, Inc. Bi-specific diabodies that are capable of binding gpA33 and CD3 and uses thereof
PL3044234T3 (en) 2013-09-13 2020-10-19 Beigene Switzerland Gmbh Anti-pd1 antibodies and their use as therapeutics and diagnostics
CA2922889A1 (en) 2013-09-17 2015-03-26 Genentech, Inc. Methods of using anti-lgr5 antibodies
CN104436157A (en) 2013-09-23 2015-03-25 恩金生物有限公司 Influenza vaccine and therapy
ES2900425T3 (en) 2013-09-25 2022-03-16 Bioverativ Therapeutics Inc Column viral inactivation methods
EP3049437A1 (en) 2013-09-27 2016-08-03 F. Hoffmann-La Roche AG Thermus thermophilus slyd fkbp domain specific antibodies
WO2015050959A1 (en) 2013-10-01 2015-04-09 Yale University Anti-kit antibodies and methods of use thereof
ES2837392T3 (en) 2013-10-02 2021-06-30 Medimmune Llc Neutralizing anti-influenza A antibodies and their uses
CA2922912A1 (en) 2013-10-11 2015-04-16 F. Hoffmann-La Roche Ag Multispecific domain exchanged common variable light chain antibodies
WO2015054670A1 (en) 2013-10-11 2015-04-16 Genentech, Inc. Nsp4 inhibitors and methods of use
WO2015057939A1 (en) 2013-10-18 2015-04-23 Biogen Idec Ma Inc. Anti-s1p4 antibodies and uses thereof
RU2016114074A (en) 2013-10-18 2017-11-23 Дженентек, Инк. ANTI-RSPO ANTIBODIES AND METHODS OF APPLICATION
EP3060685B1 (en) 2013-10-23 2019-05-01 F. Hoffmann-La Roche AG Method of predicting the response of an asthma patient to therapy
KR20220013459A (en) 2013-10-25 2022-02-04 악셀레론 파마 인코포레이티드 Endoglin peptides to treat fibrotic diseases
SI3063275T1 (en) 2013-10-31 2020-02-28 Resolve Therapeutics, Llc Therapeutic nuclease-albumin fusions and methods
RU2704228C2 (en) 2013-11-07 2019-10-24 Инсерм (Институт Насьональ Де Ла Сант Эт Де Ла Решерш Медикаль) Allosteric antibodies against human her3 non competitive for neuregulin and use thereof
CN104623637A (en) 2013-11-07 2015-05-20 健能隆医药技术（上海）有限公司 Application of IL-22 dimer in preparation of intravenous injection drugs
EP3065769A4 (en) 2013-11-08 2017-05-31 Biogen MA Inc. Procoagulant fusion compound
SG10202007189VA (en) 2013-11-21 2020-09-29 Hoffmann La Roche ANTI-alpha-SYNUCLEIN ANTIBODIES AND METHODS OF USE
US10947319B2 (en) 2013-11-27 2021-03-16 Zymeworks Inc. Bispecific antigen-binding constructs targeting HER2
BR112016012358A2 (en) 2013-12-06 2017-09-26 Dana Farber Cancer Inst Inc therapeutic peptides
CN113861293A (en) 2013-12-09 2021-12-31 爱乐科斯公司 anti-Siglec-8 antibodies and methods of use thereof
CN105814084B (en) 2013-12-13 2019-09-24 基因泰克公司 Anti-CD 33 antibody and immunoconjugates
US8980273B1 (en) 2014-07-15 2015-03-17 Kymab Limited Method of treating atopic dermatitis or asthma using antibody to IL4RA
WO2015095410A1 (en) 2013-12-17 2015-06-25 Genentech, Inc. Methods of treating cancer using pd-1 axis binding antagonists and an anti-cd20 antibody
TWI747385B (en) 2013-12-17 2021-11-21 美商建南德克公司 Anti-cd3 antibodies and methods of use
US20150190506A1 (en) 2013-12-17 2015-07-09 Genentech, Inc. Combination therapy comprising ox40 binding agonists and pd-1 axis binding antagonists
US8986691B1 (en) 2014-07-15 2015-03-24 Kymab Limited Method of treating atopic dermatitis or asthma using antibody to IL4RA
JP6841656B2 (en) 2013-12-17 2021-03-10 ジェネンテック， インコーポレイテッド Treatment of Cancer Using PD-1 Axle Antagonist and Taxane
DK3083680T3 (en) 2013-12-20 2020-03-16 Hoffmann La Roche Humanized anti-Tau (pS422) antibodies and methods for use
TWI728373B (en) 2013-12-23 2021-05-21 美商建南德克公司 Antibodies and methods of use
US11014987B2 (en) 2013-12-24 2021-05-25 Janssen Pharmaceutics Nv Anti-vista antibodies and fragments, uses thereof, and methods of identifying same
EA201691323A1 (en) 2013-12-24 2016-11-30 Янссен Фармасьютика Нв ANTI-VISTA ANTIBODIES AND FRAGMENTS
WO2015100299A1 (en) 2013-12-24 2015-07-02 Argen-X N.V. Fcrn antagonists and methods of use
RU2694981C2 (en) 2014-01-03 2019-07-18 Ф. Хоффманн-Ля Рош Аг Covalently linked conjugates chelicar-antibody against chelicar and use thereof
BR112016013849A2 (en) 2014-01-03 2017-10-10 Hoffmann La Roche bispecific antihapten / blood-brain barrier receptor conjugates, their uses, and pharmaceutical formulation
MX2016008191A (en) 2014-01-03 2017-11-16 Hoffmann La Roche Covalently linked polypeptide toxin-antibody conjugates.
EP3092251B1 (en) 2014-01-06 2021-01-20 F. Hoffmann-La Roche AG Monovalent blood brain barrier shuttle modules
CN117106095A (en) 2014-01-10 2023-11-24 比奥贝拉蒂治疗公司 Factor VIII chimeric proteins and uses thereof
MX2016008539A (en) 2014-01-15 2016-09-26 Hoffmann La Roche Fc-region variants with modified fcrn- and maintained protein a-binding properties.
KR20160111469A (en) 2014-01-24 2016-09-26 제넨테크, 인크. Methods of using anti-steap1 antibodies and immunoconjugates
ES2808340T3 (en) 2014-01-24 2021-02-26 Ngm Biopharmaceuticals Inc Antibodies that bind to the beta klotho 2 domain and procedures for using them
AU2015214264B2 (en) 2014-02-04 2018-12-20 Curis, Inc. Mutant Smoothened and methods of using the same
CN106456729A (en) 2014-02-08 2017-02-22 豪夫迈·罗氏有限公司 Methods of treating alzheimer's disease
EP3718563A1 (en) 2014-02-08 2020-10-07 F. Hoffmann-La Roche AG Methods of treating alzheimer's disease
CA3045124A1 (en) 2014-02-12 2015-08-20 Yvonne CHINN Anti-jagged1 antibodies and methods of use
KR20160124165A (en) 2014-02-21 2016-10-26 제넨테크, 인크. Anti-il-13/il-17 bispecific antibodies and uses thereof
JP6825909B2 (en) 2014-02-28 2021-02-03 アラコス インコーポレイテッド Methods and Compositions for Treating SIGLEC-8 Related Diseases
WO2015134671A1 (en) 2014-03-04 2015-09-11 Targeson, Inc. Molecular imaging contrast agents and uses thereof
NZ711451A (en) 2014-03-07 2016-05-27 Alexion Pharma Inc Anti-c5 antibodies having improved pharmacokinetics
EA201691827A1 (en) 2014-03-12 2017-01-30 Новартис Аг SPECIFIC PLOTS FOR MODIFICATION OF ANTIBODIES WITH THE PURPOSE OF OBTAINING IMMUNOCONJUGATES
CA3124243A1 (en) 2014-03-14 2015-09-17 Dana-Farber Cancer Institute, Inc. Vaccine compositions and methods for restoring nkg2d pathway function against cancers
PL3116999T3 (en) 2014-03-14 2021-12-27 F.Hoffmann-La Roche Ag Methods and compositions for secretion of heterologous polypeptides
DK3129067T3 (en) 2014-03-19 2023-03-27 Genzyme Corp SITE-SPECIFIC GLYCOMODIFICATION OF TARGETING MOBILITIES
WO2015140591A1 (en) 2014-03-21 2015-09-24 Nordlandssykehuset Hf Anti-cd14 antibodies and uses thereof
EP3119812B1 (en) 2014-03-21 2020-04-29 X-Body, Inc. Bi-specific antigen-binding polypeptides
BR112016021383A2 (en) 2014-03-24 2017-10-03 Genentech Inc METHOD TO IDENTIFY A PATIENT WITH CANCER WHO IS LIKE OR LESS LIKELY TO RESPOND TO TREATMENT WITH A CMET ANTAGONIST, METHOD TO IDENTIFY A PATIENT WITH PREVIOUSLY TREATED CANCER, METHOD TO DETERMINE THE EXPRESSION OF THE HGF BIOMARKER, ANTI-C-MET ANTAGONIST AND ITS USE, DIAGNOSTIC KIT AND ITS PREPARATION METHOD
JP6637439B2 (en) 2014-03-31 2020-01-29 ジェネンテック， インコーポレイテッド Anti-OX40 antibody and method of use
WO2015153514A1 (en) 2014-03-31 2015-10-08 Genentech, Inc. Combination therapy comprising anti-angiogenesis agents and ox40 binding agonists
WO2015153916A1 (en) 2014-04-04 2015-10-08 Bionomics, Inc. Humanized antibodies that bind lgr5
ES2879804T3 (en) 2014-04-21 2021-11-23 D&D Pharmatech Inc TRAIL receptor agonists for the treatment of fibrotic diseases
TW201622746A (en) 2014-04-24 2016-07-01 諾華公司 Methods of improving or accelerating physical recovery after surgery for hip fracture
WO2015164615A1 (en) 2014-04-24 2015-10-29 University Of Oslo Anti-gluten antibodies and uses thereof
RU2728703C2 (en) 2014-05-02 2020-07-31 Рисерч Инститьют Эт Нэшнуайд Чилдрен`С Хоспитал Compositions and methods for anti-lyst immunomodulation
MX2016015162A (en) 2014-05-22 2017-03-03 Genentech Inc Anti-gpc3 antibodies and immunoconjugates.
JP2017524371A (en) 2014-05-23 2017-08-31 ジェネンテック， インコーポレイテッド MIT biomarkers and methods of use
SI3148579T1 (en) 2014-05-28 2021-07-30 Agenus Inc. Anti-gitr antibodies and methods of use thereof
EA035419B9 (en) 2014-05-29 2020-08-07 Мэкроудженикс, Инк. Tri-specific binding molecules and methods of use thereof
SG11201609959UA (en) 2014-05-30 2016-12-29 Henlius Biotech Co Ltd Anti-epidermal growth factor receptor (egfr) antibodies
PL3151921T3 (en) 2014-06-06 2020-02-28 Bristol-Myers Squibb Company Antibodies against glucocorticoid-induced tumor necrosis factor receptor (gitr) and uses thereof
RU2016148616A (en) 2014-06-11 2018-07-18 Дженентек, Инк. ANTI-LGR5 ANTIBODIES AND THEIR APPLICATION
JP6997619B2 (en) 2014-06-11 2022-01-17 キャシー・エイ・グリーン Use of VISTA agonists and VISTA antagonists for suppression or enhancement of humoral immunity
JP2017517552A (en) 2014-06-13 2017-06-29 ジェネンテック， インコーポレイテッド Treatment and prevention of anticancer drug resistance
US20170291939A1 (en) 2014-06-25 2017-10-12 Novartis Ag Antibodies specific for il-17a fused to hyaluronan binding peptide tags
AR100978A1 (en) 2014-06-26 2016-11-16 Hoffmann La Roche ANTI-Tau HUMANIZED ANTIBODY BRAIN LAUNCHERS (pS422) AND USES OF THE SAME
RU2705299C2 (en) 2014-06-26 2019-11-06 Ф. Хоффманн-Ля Рош Аг Antibodies against 5-bromo-2'-deoxyuridine and methods of use
US11008561B2 (en) 2014-06-30 2021-05-18 Bioverativ Therapeutics Inc. Optimized factor IX gene
WO2016000619A1 (en) 2014-07-03 2016-01-07 Beigene, Ltd. Anti-pd-l1 antibodies and their use as therapeutics and diagnostics
EP3166974A1 (en) 2014-07-11 2017-05-17 Genentech, Inc. Anti-pd-l1 antibodies and diagnostic uses thereof
US9914774B2 (en) 2014-07-11 2018-03-13 Genentech, Inc. Notch pathway inhibition
AU2015289805B2 (en) 2014-07-15 2020-06-25 Humabs Biomed Sa Neutralizing anti-influenza B antibodies and uses thereof
JP6986965B2 (en) 2014-07-22 2021-12-22 アポロミクス インコーポレイテッド Anti-PD-1 antibody
WO2016019126A1 (en) 2014-07-30 2016-02-04 The Research Foundation For The State University Of New York System and method for delivering genetic material or protein to cells
CN112999429B (en) 2014-08-01 2022-07-22 麻省理工学院 Modified alginates for anti-fibrotic materials and uses
US10786578B2 (en) 2014-08-05 2020-09-29 Novartis Ag CKIT antibody drug conjugates
CN118388646A (en) 2014-08-05 2024-07-26 中美冠科生物技术（太仓）有限公司 Anti-PD-L1 antibodies
WO2016020882A2 (en) 2014-08-07 2016-02-11 Novartis Ag Angiopoetin-like 4 (angptl4) antibodies and methods of use
ES2906216T3 (en) 2014-08-07 2022-04-13 Novartis Ag Angiopoietin-like 4 antibodies and methods of use
CA2954599A1 (en) 2014-08-12 2016-02-18 Novartis Ag Anti-cdh6 antibody drug conjugates
JO3663B1 (en) 2014-08-19 2020-08-27 Merck Sharp & Dohme Anti-lag3 antibodies and antigen-binding fragments
CN107148430B (en) 2014-08-19 2021-08-27 默沙东公司 anti-TIGIT antibody
KR102615681B1 (en) 2014-08-28 2023-12-18 바이오아트라, 인코퍼레이티드 Conditionally active chimeric antigen receptors for modified t-cells
EP3567056A1 (en) 2014-09-10 2019-11-13 F. Hoffmann-La Roche AG Galactoengineered immunoglobulin 1 antibodies
RU2017107502A (en) 2014-09-12 2018-10-12 Дженентек, Инк. ANTIBODIES AND CONJUGATES DESIGNED BY THE INTRODUCTION OF CYSTEINE
CR20170131A (en) 2014-09-12 2017-07-19 Genentech Inc ANTI-HER2 AND IMMUNOCUSED ANTIBODIES
TW201625689A (en) 2014-09-12 2016-07-16 建南德克公司 Anti-B7-H4 antibodies and immunoconjugates
BR112017004802A2 (en) 2014-09-12 2017-12-12 Genentech Inc anti-cll-1 and immunoconjugate antibodies
CA2957148A1 (en) 2014-09-17 2016-03-24 Genentech, Inc. Immunoconjugates comprising anti-her2 antibodies and pyrrolobenzodiazepines
HUE049175T2 (en) 2014-09-23 2020-09-28 Hoffmann La Roche Method of using anti-cd79b immunoconjugates
TN2017000109A1 (en) 2014-09-26 2018-07-04 Bayer Pharma AG Stabilized adrenomedullin derivatives and use thereof
WO2016054101A1 (en) 2014-09-29 2016-04-07 Duke University Bispecific molecules comprising an hiv-1 envelope targeting arm
MA41044A (en) 2014-10-08 2017-08-15 Novartis Ag COMPOSITIONS AND METHODS OF USE FOR INCREASED IMMUNE RESPONSE AND CANCER TREATMENT
PT3204425T (en) 2014-10-09 2020-12-18 Genzyme Corp Glycoengineered antibody drug conjugates
CN108064282A (en) 2014-10-14 2018-05-22 哈洛齐梅公司 Adenosine deaminase -2 (ADA2), its variant composition and use its method
US9732148B2 (en) 2014-10-16 2017-08-15 Genentech, Inc. Anti-α-synuclein antibodies and methods of use
MA41480A (en) 2014-10-17 2017-12-19 Glenmark Pharmaceuticals Sa ANTIBODIES BOUND TO CCR6 AND THEIR USES
MA41685A (en) 2014-10-17 2017-08-22 Biogen Ma Inc COPPER SUPPLEMENT FOR THE REGULATION OF GLYCOSYLATION IN A MAMMAL CELL CULTURE PROCESS
WO2016070001A1 (en) 2014-10-31 2016-05-06 Jounce Therapeutics, Inc. Methods of treating conditions with antibodies that bind b7-h4
US20160160290A1 (en) 2014-11-03 2016-06-09 Genentech, Inc. Methods and biomarkers for predicting efficacy and evaluation of an ox40 agonist treatment
SG11201703448QA (en) 2014-11-03 2017-05-30 Genentech Inc Assays for detecting t cell immune subsets and methods of use thereof
CA2961439A1 (en) 2014-11-05 2016-05-12 Genentech, Inc. Anti-fgfr2/3 antibodies and methods using same
AU2015342961B2 (en) 2014-11-05 2021-08-12 Genentech, Inc. Methods of producing two chain proteins in bacteria
MX2017005925A (en) 2014-11-05 2017-11-08 Genentech Inc Methods of producing two chain proteins in bacteria.
JP2017534633A (en) 2014-11-06 2017-11-24 ジェネンテック， インコーポレイテッド Combination therapy comprising an OX40 binding agonist and a TIGIT inhibitor
MX2017005150A (en) 2014-11-06 2017-08-08 Hoffmann La Roche Fc-region variants with modified fcrn- and protein a-binding properties.
WO2016073157A1 (en) 2014-11-06 2016-05-12 Genentech, Inc. Anti-ang2 antibodies and methods of use thereof
KR20170078677A (en) 2014-11-06 2017-07-07 에프. 호프만-라 로슈 아게 Fc-region variants with modified fcrn-binding and methods of use
TWI705976B (en) 2014-11-10 2020-10-01 美商建南德克公司 Anti-interleukin-33 antibodies and uses thereof
WO2016077369A1 (en) 2014-11-10 2016-05-19 Genentech, Inc. Animal model for nephropathy and agents for treating the same
EA201791050A1 (en) 2014-11-14 2017-09-29 Новартис Аг CONJUGATES OF ANTIBODIES AND MEDICINES
JP2017537090A (en) 2014-11-17 2017-12-14 ジェネンテック， インコーポレイテッド Combination therapy comprising OX40 binding agonist and PD-1 axis binding antagonist
CN107250158B (en) 2014-11-19 2022-03-25 基因泰克公司 Anti-transferrin receptor/anti-BACE 1 multispecific antibodies and methods of use
EP3845565A3 (en) 2014-11-19 2021-09-08 Genentech, Inc. Antibodies against bace1 and use thereof for neural disease immunotherapy
EP3221362B1 (en) 2014-11-19 2019-07-24 F.Hoffmann-La Roche Ag Anti-transferrin receptor antibodies and methods of use
EP3786182A1 (en) 2014-11-19 2021-03-03 Axon Neuroscience SE Humanized tau antibodies in alzheimer's disease
SI3221346T1 (en) 2014-11-21 2020-11-30 Bristol-Myers Squibb Company Antibodies comprising modified heavy constant regions
MX2017006624A (en) 2014-11-21 2017-08-21 Bristol Myers Squibb Co Antibodies against cd73 and uses thereof.
CN107001482B (en) 2014-12-03 2021-06-15 豪夫迈·罗氏有限公司 Multispecific antibodies
CA2969730A1 (en) 2014-12-05 2016-06-09 Immunext, Inc. Identification of vsig8 as the putative vista receptor and its use thereof to produce vista/vsig8 modulators
HUE045216T2 (en) 2014-12-05 2019-12-30 Hoffmann La Roche Anti-cd79b antibodies and methods of use
EP3229836B1 (en) 2014-12-09 2019-11-13 Institut National de la Sante et de la Recherche Medicale (INSERM) Human monoclonal antibodies against axl
JP2018502840A (en) 2014-12-10 2018-02-01 ジェネンテック， インコーポレイテッド Blood brain barrier receptor antibodies and methods of use
UY36449A (en) 2014-12-19 2016-07-29 Novartis Ag COMPOSITIONS AND METHODS FOR ANTIBODIES DIRECTED TO BMP6
UA127961C2 (en) 2014-12-19 2024-02-28 Чугей Сейяку Кабусікі Кайся ANTI-MYOSTATIN ANTIBODIES, POLYPEPTIDES CONTAINING VARIANT Fc REGIONS, AND METHODS OF USE
EP3233921B1 (en) 2014-12-19 2021-09-29 Chugai Seiyaku Kabushiki Kaisha Anti-c5 antibodies and methods of use
PT3233912T (en) 2014-12-19 2021-08-09 Regenesance B V Antibodies that bind human c6 and uses thereof
TWI708786B (en) 2014-12-23 2020-11-01 美商必治妥美雅史谷比公司 Antibodies to tigit
WO2016111947A2 (en) 2015-01-05 2016-07-14 Jounce Therapeutics, Inc. Antibodies that inhibit tim-3:lilrb2 interactions and uses thereof
GB2557389B (en) 2015-01-14 2020-12-23 Brigham & Womens Hospital Inc Treatment of cancer with anti-lap monoclonal antibodies
WO2016117346A1 (en) 2015-01-22 2016-07-28 Chugai Seiyaku Kabushiki Kaisha A combination of two or more anti-c5 antibodies and methods of use
SG11201706024YA (en) 2015-01-26 2017-08-30 Macrogenics Inc Multivalent molecules comprising dr5-binding domains
WO2016123125A1 (en) 2015-01-27 2016-08-04 The Johns Hopkins University Hypotonic hydrogel formulations for enhanced transport of active agents at mucosal surfaces
WO2016123329A2 (en) 2015-01-28 2016-08-04 Genentech, Inc. Gene expression markers and treatment of multiple sclerosis
JP2018512597A (en) 2015-02-04 2018-05-17 ジェネンテック， インコーポレイテッド Mutant smoothened and method of using the same
JP2018510842A (en) 2015-02-05 2018-04-19 中外製薬株式会社 Antibodies comprising ion concentration dependent antigen binding domains, Fc region variants, antibodies that bind to IL-8, and uses thereof
KR102669294B1 (en) 2015-02-19 2024-05-23 컴퓨젠 엘티디. Anti-PVRIG antibodies and methods of use
US10550173B2 (en) 2015-02-19 2020-02-04 Compugen, Ltd. PVRIG polypeptides and methods of treatment
WO2016135041A1 (en) 2015-02-26 2016-09-01 INSERM (Institut National de la Santé et de la Recherche Médicale) Fusion proteins and antibodies comprising thereof for promoting apoptosis
EP3265491A1 (en) 2015-03-03 2018-01-10 Xoma (Us) Llc Treatment of post-prandial hyperinsulinemia and hypoglycemia after bariatric surgery
SG10201908259WA (en) 2015-03-09 2019-10-30 Argenx Bvba Methods of reducing serum levels of fc-containing agents using fcrn antagonsits
MX2017011486A (en) 2015-03-16 2018-06-15 Genentech Inc Methods of detecting and quantifying il-13 and uses in diagnosing and treating th2-associated diseases.
WO2016146833A1 (en) 2015-03-19 2016-09-22 F. Hoffmann-La Roche Ag Biomarkers for nad(+)-diphthamide adp ribosyltransferase resistance
AU2016236270B2 (en) 2015-03-23 2022-01-27 Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts Anti-CEACAM6 antibodies and uses thereof
HUE049938T2 (en) 2015-03-23 2020-11-30 Jounce Therapeutics Inc Antibodies to icos
JP6903587B2 (en) 2015-04-03 2021-07-14 ユーリカ セラピューティックス， インコーポレイテッド Constructs targeting AFP peptide / MHC complexes and their use
US10167334B2 (en) 2015-04-03 2019-01-01 Xoma Technology Ltd. Treatment of cancer using anti-TGF-BETA and PD-1 antibodies
AU2016246695A1 (en) 2015-04-07 2017-10-26 Genentech, Inc. Antigen binding complex having agonistic activity and methods of use
DK3283508T3 (en) 2015-04-17 2021-05-31 Alpine Immune Sciences Inc Immunomodulatory Proteins with Tunable Affinities
WO2016172551A2 (en) 2015-04-24 2016-10-27 Genentech, Inc. Methods of identifying bacteria comprising binding polypeptides
JP2018520642A (en) 2015-05-01 2018-08-02 ジェネンテック， インコーポレイテッド Mask anti-CD3 antibody and method of use thereof
WO2016179194A1 (en) 2015-05-04 2016-11-10 Jounce Therapeutics, Inc. Lilra3 and method of using the same
EP3292152A1 (en) 2015-05-07 2018-03-14 Agenus Inc. Anti-ox40 antibodies and methods of use thereof
JP6963508B2 (en) 2015-05-11 2021-11-10 ジェネンテック， インコーポレイテッド Compositions and Methods for Treating Lupus Nephritis
AU2016262074A1 (en) 2015-05-12 2017-11-09 Genentech, Inc. Therapeutic and diagnostic methods for cancer
JP6770533B2 (en) 2015-05-22 2020-10-14 アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル A human monoclonal antibody fragment that inhibits both the catalytic activity of Cath-D and its binding to the LRP1 receptor.
WO2016196228A1 (en) 2015-05-29 2016-12-08 Bristol-Myers Squibb Company Antibodies against ox40 and uses thereof
PL3303619T3 (en) 2015-05-29 2020-10-05 F. Hoffmann-La Roche Ag Pd-l1 promoter methylation in cancer
WO2016196343A1 (en) 2015-05-29 2016-12-08 Genentech, Inc. Humanized anti-ebola virus glycoprotein antibodies and methods of use
PL3303632T5 (en) 2015-05-29 2023-07-03 F. Hoffmann-La Roche Ag Therapeutic and diagnostic methods for cancer
RS60441B1 (en) 2015-05-30 2020-07-31 Molecular Templates Inc De-immunized, shiga toxin a subunit scaffolds and cell-targeting molecules comprising the same
US10442854B2 (en) 2015-06-01 2019-10-15 Medimmune, Llc Neutralizing anti-influenza binding molecules and uses thereof
JP2018516933A (en) 2015-06-02 2018-06-28 ジェネンテック， インコーポレイテッド Compositions and methods for treating neurological disorders using anti-IL-34 antibodies
SG11201707815RA (en) 2015-06-05 2017-12-28 Novartis Ag Antibodies targeting bone morphogenetic protein 9 (bmp9) and methods therefor
CA2986942A1 (en) 2015-06-05 2016-12-08 Genentech, Inc. Anti-tau antibodies and methods of use
KR20180011839A (en) 2015-06-08 2018-02-02 제넨테크, 인크. Treatment of Cancer Using Anti-OX40 Antibody
TWI773646B (en) 2015-06-08 2022-08-11 美商宏觀基因股份有限公司 Lag-3-binding molecules and methods of use thereof
CA2988420A1 (en) 2015-06-08 2016-12-15 Genentech, Inc. Methods of treating cancer using anti-ox40 antibodies and pd-1 axis binding antagonists
TW201709929A (en) 2015-06-12 2017-03-16 宏觀基因股份有限公司 Combination therapy for the treatment of cancer
AR104987A1 (en) 2015-06-15 2017-08-30 Genentech Inc ANTIBODY-DRUG IMMUNOCUJADOS UNITED BY NON-PEPTIDIC LINKERS
EP3310811B1 (en) 2015-06-16 2021-06-16 Genentech, Inc. Anti-cd3 antibodies and methods of use
CN107847568B (en) 2015-06-16 2022-12-20 豪夫迈·罗氏有限公司 anti-CLL-1 antibodies and methods of use
CN107849132B (en) 2015-06-16 2022-03-08 豪夫迈·罗氏有限公司 Humanized and affinity matured antibodies to FcRH5 and methods of use
MX2017016353A (en) 2015-06-17 2018-05-02 Genentech Inc Methods of treating locally advanced or metastatic breast cancers using pd-1 axis binding antagonists and taxanes.
EP3310813A1 (en) 2015-06-17 2018-04-25 Novartis AG Antibody drug conjugates
AU2016278239B9 (en) 2015-06-17 2022-08-11 Allakos Inc. Methods and compositions for treating fibrotic diseases
WO2016205531A2 (en) 2015-06-17 2016-12-22 Genentech, Inc. Anti-her2 antibodies and methods of use
MX2017016647A (en) 2015-06-24 2019-04-25 Janssen Pharmaceutica Nv Anti-vista antibodies and fragments.
CA2985718A1 (en) 2015-06-24 2016-12-29 F. Hoffmann-La Roche Ag Anti-transferrin receptor antibodies with tailored affinity
AR105089A1 (en) 2015-06-24 2017-09-06 Hoffmann La Roche ANTI-TAU ANTIBODIES (pS422) HUMANIZED AND USED METHODS
JOP20200312A1 (en) 2015-06-26 2017-06-16 Novartis Ag Factor xi antibodies and methods of use
ES2898065T3 (en) 2015-06-29 2022-03-03 Ventana Med Syst Inc Materials and Procedures for Performing Histochemical Assays for Human Proepiregulin and Amphiregulin
BR112017027736A2 (en) 2015-06-29 2018-10-09 Genentech Inc anti-cd20 type ii antibody for use in organ transplantation
WO2017005847A1 (en) 2015-07-07 2017-01-12 INSERM (Institut National de la Santé et de la Recherche Médicale) Antibodies having specificity to myosin 18a and uses thereof
CA2991980A1 (en) 2015-07-13 2017-01-19 Compugen Ltd. Hide1 compositions and methods
JP6937737B2 (en) * 2015-07-24 2021-09-22 グリックニック インコーポレイテッド Fusion protein of human protein fragments for making higher multimerized immunoglobulin FC compositions with improved complement fixation
TWI833183B (en) 2015-07-30 2024-02-21 美商宏觀基因股份有限公司 Pd-1-binding molecules and methods of use thereof
HUE048939T2 (en) 2015-08-03 2020-09-28 Engmab Sarl Monoclonal antibodies against human b cell maturation antigen (bcma)
KR20180035852A (en) 2015-08-03 2018-04-06 노파르티스 아게 Methods for treating FGF21-associated disorders
TWI741992B (en) 2015-08-03 2021-10-11 美商百歐維拉提夫治療公司 Factor ix fusion proteins and methods of making and using same
US9884900B2 (en) 2015-08-04 2018-02-06 Acceleron Pharma Inc. Methods for treating Janus kinase-associated disorders by administering soluble transforming growth factor beta type II receptor
US10253101B2 (en) 2015-08-06 2019-04-09 Xoma (Us) Llc Antibody fragments against the insulin receptor and uses thereof to treat hypoglycemia
CN105384825B (en) 2015-08-11 2018-06-01 南京传奇生物科技有限公司 A kind of bispecific chimeric antigen receptor and its application based on single domain antibody
EP3932953A1 (en) 2015-08-28 2022-01-05 F. Hoffmann-La Roche AG Anti-hypusine antibodies and uses thereof
US10323091B2 (en) 2015-09-01 2019-06-18 Agenus Inc. Anti-PD-1 antibodies and methods of use thereof
CN114507281A (en) 2015-09-09 2022-05-17 诺华股份有限公司 Thymic Stromal Lymphopoietin (TSLP) -binding molecules and methods of using the same
AU2016320748B2 (en) 2015-09-09 2019-05-02 Novartis Ag Thymic stromal lymphopoietin (TSLP)-binding antibodies and methods of using the antibodies
US20190022092A1 (en) 2015-09-15 2019-01-24 Acerta Pharma B.V. Therapeutic Combinations of a BTK Inhibitor and a GITR Binding Molecule, a 4-1BB Agonist, or an OX40 Agonist
US9862760B2 (en) 2015-09-16 2018-01-09 Novartis Ag Polyomavirus neutralizing antibodies
JP6904947B2 (en) 2015-09-22 2021-07-21 スプリング バイオサイエンス コーポレーション Anti-OX40 antibody and its diagnostic applications
KR20180053315A (en) 2015-09-23 2018-05-21 제넨테크, 인크. Optimized variants of anti-VEGF antibodies
BR112018005931A2 (en) 2015-09-24 2018-10-09 Abvitro Llc hiv antibody compositions and methods of use
IL298355A (en) 2015-09-25 2023-01-01 Genentech Inc Anti-tigit antibodies and methods of use
BR112018006237A2 (en) 2015-09-29 2018-10-09 Celgene Corp pd-1 binding proteins and methods of using them
AR106188A1 (en) 2015-10-01 2017-12-20 Hoffmann La Roche ANTI-CD19 HUMANIZED HUMAN ANTIBODIES AND METHODS OF USE
EP3356406A1 (en) 2015-10-02 2018-08-08 H. Hoffnabb-La Roche Ag Bispecific anti-human cd20/human transferrin receptor antibodies and methods of use
EP3150636A1 (en) 2015-10-02 2017-04-05 F. Hoffmann-La Roche AG Tetravalent multispecific antibodies
AR106189A1 (en) 2015-10-02 2017-12-20 Hoffmann La Roche BIESPECTIFIC ANTIBODIES AGAINST HUMAN A-b AND THE HUMAN TRANSFERRINE RECEIVER AND METHODS OF USE
MA43345A (en) 2015-10-02 2018-08-08 Hoffmann La Roche PYRROLOBENZODIAZEPINE ANTIBODY-DRUG CONJUGATES AND METHODS OF USE
US11891440B2 (en) * 2015-10-05 2024-02-06 Circle33 Llc Antibodies with improved stability to intestinal digestion, polynucleotides thereof and methods of use thereof to treat disease
CN108431038A (en) 2015-10-06 2018-08-21 豪夫迈·罗氏有限公司 The method for treating multiple sclerosis
MA43354A (en) 2015-10-16 2018-08-22 Genentech Inc CONJUGATE DRUG CONJUGATES WITH CLOUDY DISULPHIDE
WO2017066714A1 (en) 2015-10-16 2017-04-20 Compugen Ltd. Anti-vsig1 antibodies and drug conjugates
MA45326A (en) 2015-10-20 2018-08-29 Genentech Inc CALICHEAMICIN-ANTIBODY-DRUG CONJUGATES AND METHODS OF USE
US10604577B2 (en) 2015-10-22 2020-03-31 Allakos Inc. Methods and compositions for treating systemic mastocytosis
AU2016342269A1 (en) 2015-10-22 2018-03-29 Jounce Therapeutics, Inc. Gene signatures for determining icos expression
KR20180069065A (en) 2015-10-29 2018-06-22 에프. 호프만-라 로슈 아게 Anti-Mutant FC-Site Antibodies and Methods of Use
EP3184547A1 (en) 2015-10-29 2017-06-28 F. Hoffmann-La Roche AG Anti-tpbg antibodies and methods of use
JO3555B1 (en) 2015-10-29 2020-07-05 Merck Sharp & Dohme Antibody neutralizing human respiratory syncytial virus
MA44334A (en) 2015-10-29 2018-09-05 Novartis Ag ANTIBODY CONJUGATES INCLUDING A TOLL-TYPE RECEPTOR AGONIST
US10421821B2 (en) 2015-10-30 2019-09-24 Genentech, Inc. Anti-HtrA1 antibodies and methods of use thereof
WO2017075173A2 (en) 2015-10-30 2017-05-04 Genentech, Inc. Anti-factor d antibodies and conjugates
RU2018120104A (en) 2015-11-01 2019-12-02 Массачусетс Инститьют Оф Текнолоджи MODIFIED ALGINATES AS ANTI-FIBROUS MATERIALS AND THEIR APPLICATION
WO2017079768A1 (en) 2015-11-08 2017-05-11 Genentech, Inc. Methods of screening for multispecific antibodies
US11213586B2 (en) 2015-11-19 2022-01-04 Bristol-Myers Squibb Company Antibodies against glucocorticoid-induced tumor necrosis factor receptor (GITR)
CN108602885B (en) 2015-11-30 2022-05-24 百时美施贵宝公司 Anti-human IP-10 antibodies and uses thereof
EP3383908A1 (en) 2015-12-02 2018-10-10 Stsciences, Inc. Antibodies specific to glycosylated btla (b- and t- lymphocyte attenuator)
JP7089470B2 (en) 2015-12-02 2022-06-22 アジェナス インコーポレイテッド Antibodies and how to use them
KR20180086246A (en) 2015-12-02 2018-07-30 주식회사 에스티큐브앤컴퍼니 Antibodies and molecules that immunospecifically bind to BTN1A1 and their therapeutic uses
PE20181358A1 (en) 2015-12-04 2018-08-22 Novartis Ag COMPOSITIONS OF ANTIBODY GRAFTED WITH CYTOKINE AND METHODS FOR ITS USE IN IMMUNOREGULATION
EP3389714A4 (en) 2015-12-14 2019-11-13 MacroGenics, Inc. Bispecific molecules having immunoreactivity with pd-1 and ctla-4, and methods of use thereof
EP3399989B1 (en) 2015-12-16 2023-08-09 Merck Sharp & Dohme LLC Anti-lag3 antibodies and antigen-binding fragments
EA038551B1 (en) 2015-12-17 2021-09-14 Дзе Джонс Хопкинс Юниверсити Method for treating or preventing systemic sclerosis
WO2017103895A1 (en) 2015-12-18 2017-06-22 Novartis Ag Antibodies targeting cd32b and methods of use thereof
HUE065073T2 (en) 2015-12-18 2024-04-28 Chugai Pharmaceutical Co Ltd Anti-c5 antibodies and methods of use
KR20180097615A (en) 2016-01-08 2018-08-31 에프. 호프만-라 로슈 아게 Methods for the treatment of CEA-positive cancers using PD-1 axis-binding antagonists and anti-CEA / anti-CD3 bispecific antibodies
BR112018014762A2 (en) 2016-01-20 2018-12-26 Genentech Inc method of treating (early) alzheimer's disease
WO2017129763A1 (en) 2016-01-28 2017-08-03 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods and pharmaceutical compositions for the treatment of signet ring cell gastric cancer
JP6902040B2 (en) 2016-01-28 2021-07-14 アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル How to Increase the Efficacy of Immune Checkpoint Inhibitors
EP4035681A1 (en) 2016-01-28 2022-08-03 Institut National de la Santé et de la Recherche Médicale (INSERM) Methods and pharmaceutical composition for the treatment of cancer
SG10201913278PA (en) 2016-02-01 2020-02-27 Bioverativ Therapeutics Inc Optimized factor viii genes
EP3411396A1 (en) 2016-02-04 2018-12-12 Curis, Inc. Mutant smoothened and methods of using the same
BR112018016461A2 (en) 2016-02-12 2019-10-01 Janssen Pharmaceutica Nv antibodies and anti-sight fragments, their uses and their identification methods
WO2017144668A1 (en) 2016-02-26 2017-08-31 INSERM (Institut National de la Santé et de la Recherche Médicale) Antibodies having specificity for btla and uses thereof
CN109196121B (en) 2016-02-29 2022-01-04 基因泰克公司 Methods for treatment and diagnosis of cancer
CA3016187A1 (en) 2016-03-04 2017-09-08 Bristol-Myers Squibb Company Combination therapy with anti-cd73 antibodies
WO2017159699A1 (en) 2016-03-15 2017-09-21 Chugai Seiyaku Kabushiki Kaisha Methods of treating cancers using pd-1 axis binding antagonists and anti-gpc3 antibodies
MX2018011219A (en) 2016-03-16 2019-01-10 Merrimack Pharmaceuticals Inc Engineered trail for cancer therapy.
AU2017239038B2 (en) 2016-03-22 2024-06-27 Bionomics Inc Administration of an anti-LGR5 monoclonal antibody
KR102438140B1 (en) 2016-03-22 2022-08-31 엥스띠뛰 나씨오날 드 라 쌍떼 에 드 라 흐쉐르슈 메디깔 Humanized anti-claudin-1 antibodies and uses thereof
WO2017165734A1 (en) 2016-03-25 2017-09-28 Genentech, Inc. Multiplexed total antibody and antibody-conjugated drug quantification assay
MA45328A (en) 2016-04-01 2019-02-06 Avidity Biosciences Llc NUCLEIC ACID-POLYPEPTIDE COMPOSITIONS AND USES THEREOF
EP3439687A1 (en) 2016-04-07 2019-02-13 The Johns Hopkins University Compositions and methods for treating pancreatitis and pain with death receptor agonists
WO2017180864A1 (en) 2016-04-14 2017-10-19 Genentech, Inc. Anti-rspo3 antibodies and methods of use
SG11201808783XA (en) 2016-04-15 2018-11-29 Alpine Immune Sciences Inc Cd80 variant immunomodulatory proteins and uses thereof
EP3443350B1 (en) 2016-04-15 2020-12-09 H. Hoffnabb-La Roche Ag Methods for monitoring and treating cancer
WO2017181148A2 (en) 2016-04-15 2017-10-19 Alpine Immune Sciences, Inc. Icos ligand variant immunomodulatory proteins and uses thereof
WO2017181079A2 (en) 2016-04-15 2017-10-19 Genentech, Inc. Methods for monitoring and treating cancer
WO2017180842A1 (en) 2016-04-15 2017-10-19 Bioatla, Llc Anti-axl antibodies, antibody fragments and their immunoconjugates and uses thereof
EP3448412A4 (en) 2016-04-15 2020-03-25 Immunext Inc. Anti-human vista antibodies and use thereof
ES2930351T3 (en) 2016-04-15 2022-12-09 Evive Biotechnology Shanghai Ltd An IL-22 dimer for use in the treatment of necrotizing enterocolitis
JP7002467B2 (en) 2016-04-15 2022-01-20 マクロジェニクス，インコーポレーテッド New B7-H3 binding molecule, its antibody drug conjugate, and how to use it
US11514331B2 (en) 2016-04-27 2022-11-29 Massachusetts Institute Of Technology Sequence-controlled polymer random access memory storage
WO2017189870A1 (en) 2016-04-27 2017-11-02 Massachusetts Institute Of Technology Stable nanoscale nucleic acid assemblies and methods thereof
WO2017189724A1 (en) 2016-04-27 2017-11-02 Novartis Ag Antibodies against growth differentiation factor 15 and uses thereof
UA123323C2 (en) 2016-05-02 2021-03-17 Ф. Хоффманн-Ля Рош Аг The contorsbody - a single chain target binder
SG11201809793UA (en) 2016-05-09 2018-12-28 Bristol Myers Squibb Co Tl1a antibodies and uses thereof
US20190151346A1 (en) 2016-05-10 2019-05-23 INSERM (Institute National de la Santé et de la Recherche Médicale) Combinations therapies for the treatment of cancer
WO2017194441A1 (en) 2016-05-11 2017-11-16 F. Hoffmann-La Roche Ag Modified anti-tenascin antibodies and methods of use
IL262404B2 (en) 2016-05-13 2024-04-01 Bioatla Llc Anti-ror2 antibodies, antibody fragments, their immunoconjugates and uses thereof
ES2858151T3 (en) 2016-05-20 2021-09-29 Hoffmann La Roche PROTAC-Antibody Conjugates and Procedures for Use
EP3463452A1 (en) 2016-05-24 2019-04-10 Institut National de la Sante et de la Recherche Medicale (INSERM) Methods and pharmaceutical compositions for the treatment of non small cell lung cancer (nsclc) that coexists with chronic obstructive pulmonary disease (copd)
TW201802121A (en) 2016-05-25 2018-01-16 諾華公司 Reversal binding agents for anti-factor XI/XIa antibodies and uses thereof
US20170370906A1 (en) 2016-05-27 2017-12-28 Genentech, Inc. Bioanalytical analysis of site-specific antibody drug conjugates
TWI781934B (en) 2016-05-27 2022-11-01 美商艾吉納斯公司 Anti-tim-3 antibodies and methods of use thereof
WO2017214024A1 (en) 2016-06-06 2017-12-14 Genentech, Inc. Silvestrol antibody-drug conjugates and methods of use
CN110381988A (en) 2016-06-15 2019-10-25 诺华股份有限公司 Use the method for the inhibitor for treating disease of Bone Morphogenetic Protein 6 (BMP6)
CN116143918A (en) 2016-06-24 2023-05-23 豪夫迈·罗氏有限公司 Anti-polyubiquitin multispecific antibodies
JP7308034B2 (en) 2016-07-01 2023-07-13 リゾルブ セラピューティクス， エルエルシー Optimized double nuclease fusions and methods
EP3478717B1 (en) 2016-07-04 2022-01-05 F. Hoffmann-La Roche AG Novel antibody format
EP3481393B1 (en) 2016-07-05 2021-04-14 Beigene, Ltd. Combination of a pd-1 antagonist and a raf inhibitor for treating cancer
UY37325A (en) 2016-07-14 2018-01-31 Bristol Myers Squibb Company Una Corporacion Del Estado De Delaware MONOCLONAL ANTIBODIES THAT LINK TO TIM3 TO STIMULATE IMMUNE RESPONSES AND COMPOSITIONS CONTAINING THEM
WO2018014260A1 (en) 2016-07-20 2018-01-25 Nanjing Legend Biotech Co., Ltd. Multispecific antigen binding proteins and methods of use thereof
MA45715A (en) 2016-07-25 2019-05-29 Biogen Ma Inc ANTI-HSPA5 ANTIBODIES (GRP78) AND THEIR USES
US11471488B2 (en) 2016-07-28 2022-10-18 Alpine Immune Sciences, Inc. CD155 variant immunomodulatory proteins and uses thereof
US11834490B2 (en) 2016-07-28 2023-12-05 Alpine Immune Sciences, Inc. CD112 variant immunomodulatory proteins and uses thereof
CA3032120A1 (en) 2016-07-28 2018-02-01 Alpine Immune Sciences, Inc. Cd155 variant immunomodulatory proteins and uses thereof
US20190185578A1 (en) 2016-07-29 2019-06-20 Chugai Seiyaku Kabushiki Kaisha Bispecific antibody exhibiting increased alternative fviii-cofactor-function activity
BR112019001693A2 (en) 2016-07-29 2019-07-02 Ct Hospitalier Universitaire Toulouse antibodies targeting tumor-associated macrophages and their uses
NL2017267B1 (en) 2016-07-29 2018-02-01 Aduro Biotech Holdings Europe B V Anti-pd-1 antibodies
WO2018026748A1 (en) 2016-08-01 2018-02-08 Xoma (Us) Llc Parathyroid hormone receptor 1 (pth1r) antibodies and uses thereof
NL2017270B1 (en) 2016-08-02 2018-02-09 Aduro Biotech Holdings Europe B V New anti-hCTLA-4 antibodies
EP3494141A4 (en) 2016-08-03 2020-04-08 Bio-Techne Corporation Identification of vsig3/vista as a novel immune checkpoint and use thereof for immunotherapy
RU2757394C2 (en) 2016-08-03 2021-10-14 Нексткьюр, Инк. Compositions and methods for modulating the transmission of a lair signal
WO2018027204A1 (en) 2016-08-05 2018-02-08 Genentech, Inc. Multivalent and multiepitopic anitibodies having agonistic activity and methods of use
EA201990222A1 (en) 2016-08-05 2019-07-31 МЕДИММЬЮН, ЭлЭлСи ANTIBODIES TO O2 AND WAYS OF THEIR APPLICATION
TWI831965B (en) 2016-08-05 2024-02-11 日商中外製藥股份有限公司 Compositions for the treatment or prevention of IL-8 related diseases
IL264674B2 (en) 2016-08-05 2023-09-01 Allakos Inc Anti-siglec-7 antibodies for the treatment of cancer
JP7250674B2 (en) 2016-08-08 2023-04-03 エフ・ホフマン－ラ・ロシュ・アクチェンゲゼルシャフト CANCER TREATMENT AND DIAGNOSTIC METHOD
JP6993403B2 (en) 2016-08-10 2022-03-03 アジュ ユニバーシティー インダストリー－アカデミック コーオペレイション ファウンデーション Heterodimer Fc fusion cytokine and pharmaceutical composition containing it
EP3496763A1 (en) 2016-08-11 2019-06-19 Genentech, Inc. Pyrrolobenzodiazepine prodrugs and antibody conjugates thereof
EP4353747A3 (en) 2016-08-19 2024-06-26 BeiGene Switzerland GmbH Combination of zanubrutinib with an anti-cd20 or an anti-pd-1 antibody for use in treating cancer
WO2018044970A1 (en) 2016-08-31 2018-03-08 University Of Rochester Human monoclonal antibodies to human endogenous retrovirus k envelope (herv-k) and uses thereof
WO2018041989A1 (en) 2016-09-02 2018-03-08 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for diagnosing and treating refractory celiac disease type 2
EP3512883A1 (en) 2016-09-13 2019-07-24 Humanigen, Inc. Epha3 antibodies for the treatment of pulmonary fibrosis
KR102469700B1 (en) 2016-09-15 2022-11-23 우니베르지테트 스튜트가르트 Antigen binding protein to HER3
SG10201607778XA (en) 2016-09-16 2018-04-27 Chugai Pharmaceutical Co Ltd Anti-Dengue Virus Antibodies, Polypeptides Containing Variant Fc Regions, And Methods Of Use
EP3512885B1 (en) 2016-09-16 2024-02-21 Shanghai Henlius Biotech, Inc. Anti-pd-1 antibodies
SG11201901950TA (en) 2016-09-19 2019-04-29 Celgene Corp Methods of treating immune disorders using pd-1 binding proteins
EP3515932B1 (en) 2016-09-19 2023-11-22 F. Hoffmann-La Roche AG Complement factor based affinity chromatography
US10766958B2 (en) 2016-09-19 2020-09-08 Celgene Corporation Methods of treating vitiligo using PD-1 binding antibodies
RU2759334C2 (en) 2016-09-21 2021-11-12 Нексткьюр, Инк. Antibodies against siglec-15 and their application methods
EP4360714A3 (en) 2016-09-21 2024-07-24 Nextcure, Inc. Antibodies for siglec-15 and methods of use thereof
CA3031589A1 (en) 2016-09-23 2018-03-29 Genentech, Inc. Uses of il-13 antagonists for treating atopic dermatitis
JOP20190055A1 (en) 2016-09-26 2019-03-24 Merck Sharp & Dohme Anti-cd27 antibodies
EP3518969A2 (en) 2016-09-28 2019-08-07 Xoma (Us) Llc Antibodies that bind interleukin-2 and uses thereof
WO2018065501A1 (en) 2016-10-05 2018-04-12 F. Hoffmann-La Roche Ag Methods for preparing antibody drug conjugates
CA3038712A1 (en) 2016-10-06 2018-04-12 Genentech, Inc. Therapeutic and diagnostic methods for cancer
WO2018068201A1 (en) 2016-10-11 2018-04-19 Nanjing Legend Biotech Co., Ltd. Single-domain antibodies and variants thereof against ctla-4
TWI773694B (en) 2016-10-11 2022-08-11 美商艾吉納斯公司 Anti-lag-3 antibodies and methods of use thereof
KR20190064636A (en) 2016-10-19 2019-06-10 메디뮨 엘엘씨 Anti-O1 antibodies and uses thereof
CN110121508A (en) 2016-10-25 2019-08-13 法国国家健康和医学研究院 Monoclonal antibody in conjunction with CD160 transmembrane isofonns
US11555076B2 (en) 2016-10-29 2023-01-17 Genentech, Inc. Anti-MIC antibodies and methods of use
TWI788307B (en) 2016-10-31 2023-01-01 美商艾歐凡斯生物治療公司 Engineered artificial antigen presenting cells for tumor infiltrating lymphocyte expansion
SG11201903857UA (en) 2016-11-02 2019-05-30 Jounce Therapeutics Inc Antibodies to pd-1 and uses thereof
WO2018083204A1 (en) 2016-11-02 2018-05-11 Engmab Sàrl Bispecific antibody against bcma and cd3 and an immunological drug for combined use in treating multiple myeloma
EP3538546A1 (en) 2016-11-14 2019-09-18 Novartis AG Compositions, methods, and therapeutic uses related to fusogenic protein minion
AU2017361081A1 (en) 2016-11-15 2019-05-23 Genentech, Inc. Dosing for treatment with anti-CD20/anti-CD3 bispecific antibodies
JOP20190100A1 (en) 2016-11-19 2019-05-01 Potenza Therapeutics Inc Anti-gitr antigen-binding proteins and methods of use thereof
EP3468991A1 (en) 2016-11-21 2019-04-17 cureab GmbH Anti-gp73 antibodies and immunoconjugates
AU2017363309A1 (en) 2016-11-23 2019-07-11 Bioverativ Therapeutics Inc. Mono- and bispecific antibodies binding to coagulation factor IX and coagulation factor X
KR20240069834A (en) 2016-12-02 2024-05-20 바이오버라티브 테라퓨틱스 인크. Methods of treating hemophilic arthropathy using chimeric clotting factors
KR20190091292A (en) 2016-12-02 2019-08-05 바이오버라티브 테라퓨틱스 인크. How to Induce Immune Resistance to Coagulation Factors
EP3551655A2 (en) 2016-12-07 2019-10-16 Genentech, Inc. Anti-tau antibodies and methods of their use
SG10201911351SA (en) 2016-12-07 2020-02-27 Genentech Inc Anti-tau antibodies and methods of use
EP3551660B1 (en) 2016-12-07 2023-09-13 Agenus Inc. Anti-ctla-4 antibodies and methods of use thereof
KR102603681B1 (en) 2016-12-07 2023-11-17 아게누스 인코포레이티드 Antibodies and methods of using them
WO2018111890A1 (en) 2016-12-12 2018-06-21 Genentech, Inc. Methods of treating cancer using anti-pd-l1 antibodies and antiandrogens
WO2018114748A1 (en) 2016-12-20 2018-06-28 F. Hoffmann-La Roche Ag Combination therapy of anti-cd20/anti-cd3 bispecific antibodies and 4-1bb (cd137) agonists
JOP20190134A1 (en) 2016-12-23 2019-06-02 Potenza Therapeutics Inc Anti-neuropilin antigen-binding proteins and methods of use thereof
IL267538B2 (en) 2016-12-23 2024-05-01 Novartis Ag Anti-factor xi/xia antibodies for use in preventing, treating, managing or reducing the risk of a thromboembolic disorder or stroke in a subject
TW201825515A (en) 2017-01-04 2018-07-16 美商伊繆諾金公司 Met antibodies and immunoconjugates and uses thereof
US11357841B2 (en) 2017-01-06 2022-06-14 Iovance Biotherapeutics, Inc. Expansion of tumor infiltrating lymphocytes with potassium channel agonists and therapeutic uses thereof
JP2020505330A (en) 2017-01-06 2020-02-20 アビディティー バイオサイエンシーズ エルエルシー Nucleic acid polypeptide compositions and methods for inducing exon skipping
MX2019007963A (en) 2017-01-06 2019-10-21 Iovance Biotherapeutics Inc Expansion of tumor infiltrating lymphocytes (tils) with tumor necrosis factor receptor superfamily (tnfrsf) agonists and therapeutic combinations of tils and tnfrsf agonists.
US20180244785A1 (en) 2017-01-09 2018-08-30 Merrimack Pharmaceuticals, Inc. Anti-fgfr antibodies and methods of use
TW201831517A (en) 2017-01-12 2018-09-01 美商優瑞科生物技術公司 Constructs targeting histone h3 peptide/mhc complexes and uses thereof
TWI774726B (en) 2017-01-25 2022-08-21 英屬開曼群島商百濟神州有限公司 Crystalline forms of (s)-7-(1-(but-2-ynoyl)piperidin-4-yl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide, preparation, and uses thereof
CR20220563A (en) 2017-01-31 2022-12-07 Chugai Pharmaceutical Co Ltd A pharmaceutical composition for use in the treatment or prevention of a c5-related disease and a method for treating or preventing a c5-related disease
JOP20190187A1 (en) 2017-02-03 2019-08-01 Novartis Ag Anti-ccr7 antibody drug conjugates
CN110520443B (en) 2017-02-08 2023-07-25 诺华股份有限公司 FGF21 mimetic antibodies and uses thereof
AU2018217816A1 (en) 2017-02-10 2019-08-15 Genentech, Inc. Anti-tryptase antibodies, compositions thereof, and uses thereof
US11142570B2 (en) 2017-02-17 2021-10-12 Bristol-Myers Squibb Company Antibodies to alpha-synuclein and uses thereof
WO2018156180A1 (en) 2017-02-24 2018-08-30 Kindred Biosciences, Inc. Anti-il31 antibodies for veterinary use
KR20190134631A (en) 2017-03-01 2019-12-04 제넨테크, 인크. How to diagnose and treat cancer
WO2018158398A1 (en) 2017-03-02 2018-09-07 INSERM (Institut National de la Santé et de la Recherche Médicale) Antibodies having specificity to nectin-4 and uses thereof
US20200087403A1 (en) 2017-03-15 2020-03-19 INSERM (Institut National de la Santé et de la Recherche Médicale) Pharmaceutical compositions for the treatment of thrombosis in patients suffering from a myeloproliferative neoplasm
SG11201907769XA (en) 2017-03-16 2019-09-27 Alpine Immune Sciences Inc Cd80 variant immunomodulatory proteins and uses thereof
CN110809581A (en) 2017-03-16 2020-02-18 高山免疫科学股份有限公司 PD-L2 variant immunomodulatory proteins and uses thereof
HRP20231382T1 (en) 2017-03-16 2024-02-16 Alpine Immune Sciences, Inc. Pd-l1 variant immunomodulatory proteins and uses thereof
KR20190138636A (en) 2017-03-22 2019-12-13 제넨테크, 인크. Optimized Antibody Compositions for the Treatment of Eye Disorders
KR102628323B1 (en) 2017-03-24 2024-01-22 노바르티스 아게 How to prevent and treat heart disease
WO2018183175A1 (en) 2017-03-28 2018-10-04 Genentech, Inc. Methods of treating neurodegenerative diseases
JOP20190203A1 (en) 2017-03-30 2019-09-03 Potenza Therapeutics Inc Anti-tigit antigen-binding proteins and methods of use thereof
WO2018185618A1 (en) 2017-04-03 2018-10-11 Novartis Ag Anti-cdh6 antibody drug conjugates and anti-gitr antibody combinations and methods of treatment
US20200024351A1 (en) 2017-04-03 2020-01-23 Jounce Therapeutics, Inc. Compositions and Methods for the Treatment of Cancer
CR20190434A (en) 2017-04-05 2019-11-01 Hoffmann La Roche Anti-lag3 antibodies
TWI796329B (en) 2017-04-07 2023-03-21 美商默沙東有限責任公司 Anti-ilt4 antibodies and antigen-binding fragments
JP7297672B2 (en) 2017-04-13 2023-06-26 アジェナス インコーポレイテッド ANTI-CD137 ANTIBODY AND METHODS OF USE THEREOF
CA3058134A1 (en) 2017-04-13 2018-10-18 Aduro Biotech Holdings, Europe B.V. Anti-sirp alpha antibodies
CN118453852A (en) 2017-04-14 2024-08-09 埃克塞里艾克西斯公司 AMHRII-binding compounds for preventing or treating cancer
CN110945128B (en) 2017-04-14 2023-11-03 代表亚利桑那大学的亚利桑那董事会 Compositions and methods for treating pulmonary fibrosis
CN110944665B (en) 2017-04-14 2024-04-19 埃克塞里艾克西斯公司 AMHRII-binding compounds for the prevention or treatment of lung cancer
TW201839400A (en) 2017-04-14 2018-11-01 美商建南德克公司 Diagnostic and therapeutic methods for cancer
US20190078160A1 (en) 2017-04-21 2019-03-14 Genentech, Inc. Use of klk5 antagonists for treatment of a disease
WO2018200586A1 (en) 2017-04-26 2018-11-01 Eureka Therapeutics, Inc. Constructs specifically recognizing glypican 3 and uses thereof
JP2020517695A (en) 2017-04-27 2020-06-18 テサロ， インコーポレイテッド Antibody drugs against lymphocyte activation gene-3 (LAG-3) and their use
AR111651A1 (en) 2017-04-28 2019-08-07 Novartis Ag CONJUGATES OF ANTIBODIES THAT INCLUDE TOLL TYPE RECEIVER AGONISTS AND COMBINATION THERAPIES
FI3618863T3 (en) 2017-05-01 2023-09-01 Agenus Inc Anti-tigit antibodies and methods of use thereof
PT3628049T (en) 2017-05-04 2023-07-03 Acceleron Pharma Inc Tgf-beta receptor type ii fusion proteins and uses thereof
EP3618868A4 (en) 2017-05-05 2021-02-24 Allakos Inc. Methods and compositions for treating allergic ocular diseases
CA3062874A1 (en) 2017-05-10 2018-11-16 Iovance Biotherapeutics, Inc. Expansion of tumor infiltrating lymphocytes from liquid tumors and therapeutic uses thereof
WO2018209125A1 (en) 2017-05-10 2018-11-15 Fred Hutchinson Cancer Research Center Epstein barr virus antibodies, vaccines, and uses of the same
EP3625251A1 (en) 2017-05-15 2020-03-25 University Of Rochester Broadly neutralizing anti-influenza monoclonal antibody and uses thereof
CN111107868A (en) 2017-05-24 2020-05-05 诺华股份有限公司 Antibody cytokine transplantation proteins and methods of use
JOP20190271A1 (en) 2017-05-24 2019-11-21 Novartis Ag Antibody-cytokine engrafted proteins and methods of use for immune related disorders
BR112019024556A2 (en) 2017-05-24 2020-06-23 Novartis Ag GRAFT PROTEINS WITH ANTIBODY CYTOKIN AND METHODS FOR USE IN CANCER TREATMENT
WO2018215937A1 (en) 2017-05-24 2018-11-29 Novartis Ag Interleukin-7 antibody cytokine engrafted proteins and methods of use in the treatment of cancer
KR20220167342A (en) 2017-05-25 2022-12-20 브리스톨-마이어스 스큅 컴퍼니 Antibodies comprising modified heavy constant regions
EP3630834A1 (en) 2017-05-31 2020-04-08 STCube & Co., Inc. Methods of treating cancer using antibodies and molecules that immunospecifically bind to btn1a1
EP3630835A1 (en) 2017-05-31 2020-04-08 STCube & Co., Inc. Antibodies and molecules that immunospecifically bind to btn1a1 and the therapeutic uses thereof
WO2018220446A1 (en) 2017-06-01 2018-12-06 Compugen Ltd. Triple combination antibody therapies
WO2018226671A1 (en) 2017-06-06 2018-12-13 Stcube & Co., Inc. Methods of treating cancer using antibodies and molecules that bind to btn1a1 or btn1a1-ligands
UY37758A (en) 2017-06-12 2019-01-31 Novartis Ag METHOD OF MANUFACTURING OF BIESPECTIFIC ANTIBODIES, BISPECTIFIC ANTIBODIES AND THERAPEUTIC USE OF SUCH ANTIBODIES
WO2018229706A1 (en) 2017-06-16 2018-12-20 Novartis Ag Combination therapy for the treatment of cancer
WO2018229715A1 (en) 2017-06-16 2018-12-20 Novartis Ag Compositions comprising anti-cd32b antibodies and methods of use thereof
WO2018236889A2 (en) 2017-06-19 2018-12-27 Massachusetts Institute Of Technology Automated methods for scalable, parallelized enzymatic biopolymer synthesis and modification using microfluidic devices
US20190062428A1 (en) 2017-06-19 2019-02-28 Surface Oncology, Inc. Combination of anti-cd47 antibodies and cell death-inducing agents, and uses thereof
CR20190532A (en) 2017-06-20 2020-01-10 Amgen Inc Method of treating or ameliorating metabolic disorders using binding proteins for gastric inhibitory peptide receptor (gipr) in combination with glp-1 agonists
WO2018234843A1 (en) 2017-06-22 2018-12-27 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods and pharmaceutical compositions for the treatment of fibrosis with agents capable of inhibiting the activation of mucosal-associated invariant t (mait) cells
EP3645569A4 (en) 2017-06-26 2021-03-24 BeiGene, Ltd. Immunotherapy for hepatocellular carcinoma
US20200181271A1 (en) 2017-06-28 2020-06-11 Novartis Ag Methods for preventing and treating urinary incontinence
WO2019011918A1 (en) 2017-07-10 2019-01-17 International - Drug - Development - Biotech Treatment of b cell malignancies using afucosylated pro-apoptotic anti-cd19 antibodies in combination with anti cd20 antibodies or chemotherapeutics
MX2020000342A (en) 2017-07-11 2020-08-17 Compass Therapeutics Llc Agonist antibodies that bind human cd137 and uses thereof.
KR20240006698A (en) 2017-07-21 2024-01-15 제넨테크, 인크. Therapeutic and diagnostic methods for cancer
GB201711809D0 (en) 2017-07-21 2017-09-06 Governors Of The Univ Of Alberta Antisense oligonucleotide
EP3658173A1 (en) 2017-07-25 2020-06-03 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods and pharmaceutical compositions for modulating monocytopoiesis
PL3658589T3 (en) 2017-07-26 2024-03-18 Forty Seven, Inc. Anti-sirp-alpha antibodies and related methods
CN111315411B (en) 2017-07-27 2023-02-28 瑞颂医药公司 High concentration anti-C5 antibody formulations
SG11202000764RA (en) 2017-08-09 2020-02-27 Bioverativ Therapeutics Inc Nucleic acid molecules and uses thereof
KR20200057701A (en) * 2017-08-15 2020-05-26 킨드레드 바이오사이언시스, 인코포레이티드 Veterinary IGG FC variant
EP3684811A2 (en) 2017-08-17 2020-07-29 Massachusetts Institute of Technology Multiple specificity binders of cxc chemokines and uses thereof
BR112020003670A2 (en) 2017-08-22 2020-09-01 Sanabio, Llc soluble interferon receptors and their uses
JP7437301B2 (en) 2017-08-25 2024-02-22 ファイヴ プライム セラピューティクス インク B7-H4 antibody and its usage
US11208495B2 (en) 2017-08-28 2021-12-28 Angiex, Inc. Anti-TM4SF1 antibodies and methods of using same
WO2019051164A1 (en) 2017-09-07 2019-03-14 Augusta University Research Institute, Inc. Antibodies to programmed cell death protein 1
JP7382922B2 (en) 2017-09-20 2023-11-17 中外製薬株式会社 Dosing regimen for combination therapy using PD-1 system binding antagonists and GPC3 targeting agents
EP3684471A1 (en) 2017-09-20 2020-07-29 Institut National de la Sante et de la Recherche Medicale (INSERM) Methods and pharmaceutical compositions for modulating autophagy
KR20200133323A (en) 2017-09-21 2020-11-27 임체크 테라퓨틱스 에스에이에스 Antibodies with specificity for BTN2 and uses thereof
US11230601B2 (en) 2017-10-10 2022-01-25 Tilos Therapeutics, Inc. Methods of using anti-lap antibodies
EP3694870A1 (en) 2017-10-10 2020-08-19 Alpine Immune Sciences, Inc. Ctla-4 variant immunomodulatory proteins and uses thereof
CA3078974A1 (en) 2017-10-12 2019-04-18 Immunowake Inc. Vegfr-antibody light chain fusion protein
JP7438106B2 (en) 2017-10-14 2024-02-26 シートムエックス セラピューティクス，インコーポレイテッド Antibodies, activatable antibodies, bispecific antibodies, and bispecific activatable antibodies and methods of use thereof
EA202090974A1 (en) 2017-10-18 2020-08-05 Элпайн Иммьюн Сайенсиз, Инк. VARIANT IMMUNOMODULATING PROTEINS OF ICOS LIGAND AND ACCOMPANYING COMPOSITIONS AND METHODS
CA3078157A1 (en) 2017-10-20 2019-04-25 F.Hoffmann-La Roche Ag Method for generating multispecific antibodies from monospecific antibodies
US20210040205A1 (en) 2017-10-25 2021-02-11 Novartis Ag Antibodies targeting cd32b and methods of use thereof
JP7438942B2 (en) 2017-10-30 2024-02-27 エフ． ホフマン－ラ ロシュ アーゲー Methods for in vivo generation of multispecific antibodies from monospecific antibodies
WO2019089753A2 (en) 2017-10-31 2019-05-09 Compass Therapeutics Llc Cd137 antibodies and pd-1 antagonists and uses thereof
KR20200075860A (en) 2017-11-06 2020-06-26 제넨테크, 인크. How to diagnose and treat cancer
US10940171B2 (en) 2017-11-10 2021-03-09 Massachusetts Institute Of Technology Microbial production of pure single stranded nucleic acids
WO2019100052A2 (en) 2017-11-20 2019-05-23 Compass Therapeutics Llc Cd137 antibodies and tumor antigen-targeting antibodies and uses thereof
BR112020010016A2 (en) 2017-11-22 2020-11-10 Novartis Ag reversal binding agents for anti-factor xi / xia antibodies and their uses
CA3083118A1 (en) 2017-11-22 2019-05-31 Iovance Biotherapeutics, Inc. Expansion of peripheral blood lymphocytes (pbls) from peripheral blood
WO2019101995A1 (en) 2017-11-27 2019-05-31 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods and pharmaceutical compositions for cardiac regeneration
CN111801334B (en) 2017-11-29 2023-06-09 百济神州瑞士有限责任公司 Treatment of indolent or invasive B-cell lymphomas using combinations comprising BTK inhibitors
US11433132B2 (en) 2017-12-01 2022-09-06 Novartis Ag Polyomavirus neutralizing antibodies
MX2020005662A (en) 2017-12-01 2020-08-20 Pfizer Anti-cxcr5 antibodies and compositions and uses thereof.
CN118667812A (en) 2017-12-06 2024-09-20 艾维迪提生物科学公司 Compositions and methods for treating muscular atrophy and tonic muscular dystrophy
MA51032A (en) 2017-12-08 2021-03-17 Argenx Bvba USE OF FCRN ANTAGONISTS FOR THE TREATMENT OF GENERALIZED SEVERE MYASTHENIA
JP2021508104A (en) 2017-12-15 2021-02-25 アイオバンス バイオセラピューティクス，インコーポレイテッド Systems and methods for determining beneficial administration of tumor-infiltrating lymphocytes and their use, as well as beneficial administration of tumor-infiltrating lymphocytes and their use.
US11773171B2 (en) 2017-12-19 2023-10-03 Surrozen Operating, Inc. WNT surrogate molecules and uses thereof
WO2019126399A1 (en) 2017-12-19 2019-06-27 Surrozen, Inc. Anti-frizzled antibodies and methods of use
WO2019126401A1 (en) 2017-12-19 2019-06-27 Surrozen, Inc. Anti-lrp5/6 antibodies and methods of use
WO2019126536A1 (en) 2017-12-20 2019-06-27 Alexion Pharmaceuticals Inc. Humanized anti-cd200 antibodies and uses thereof
US20210087267A1 (en) 2017-12-20 2021-03-25 Alexion Pharmaceuticals, Inc. Liquid formulations of anti-cd200 antibodies
AU2018389111A1 (en) 2017-12-22 2020-06-18 Jounce Therapeutics, Inc. Antibodies to LILRB2
WO2019126472A1 (en) 2017-12-22 2019-06-27 Genentech, Inc. Use of pilra binding agents for treatment of a disease
SG11202005273XA (en) 2017-12-28 2020-07-29 Nanjing Legend Biotech Co Ltd Antibodies and variants thereof against pd-l1
JP7369127B2 (en) 2017-12-28 2023-10-25 ナンジン レジェンド バイオテック カンパニー，リミテッド Single domain antibodies against TIGIT and variants thereof
JP7436365B2 (en) 2017-12-29 2024-02-21 エフ． ホフマン－ラ ロシュ アーゲー Anti-VEGF antibodies and methods of use
SG11202006148UA (en) 2018-01-03 2020-07-29 Alpine Immune Sciences Inc Multi-domain immunomodulatory proteins and methods of use thereof
CA3087537A1 (en) 2018-01-04 2019-07-11 Jan-willem THEUNISSEN Anti-tissue factor antibodies, antibody-drug conjugates, and related methods
CN111886247A (en) 2018-01-05 2020-11-03 Ac免疫有限公司 Misfolded TDP-43 binding molecules
KR20200108870A (en) 2018-01-12 2020-09-21 브리스톨-마이어스 스큅 컴퍼니 Antibodies to TIM3 and uses thereof
CA3084518A1 (en) 2018-01-15 2019-07-18 Nanjing Legend Biotech Co., Ltd. Single-domain antibodies and variants thereof against pd-1
US20200339686A1 (en) 2018-01-16 2020-10-29 Lakepharma, Inc. Bispecific antibody that binds cd3 and another target
WO2019143837A1 (en) 2018-01-17 2019-07-25 Apexigen, Inc. Anti-pd-l1 antibodies and methods of use
WO2019147670A1 (en) 2018-01-23 2019-08-01 Nextcure, Inc. B7-h4 antibodies and methods of use thereof
US20210054064A1 (en) 2018-01-24 2021-02-25 INSERM (Institut National de la Santé et de la Recherche Médicale Antagonists of il-33 for use in methods for preventing ischemia reperfusion injusry in an organ
EP3743437A1 (en) 2018-01-26 2020-12-02 F. Hoffmann-La Roche AG Il-22 fc fusion proteins and methods of use
MX2020007628A (en) 2018-01-26 2020-10-12 Genentech Inc Compositions and methods of use.
US20210038744A1 (en) 2018-02-01 2021-02-11 Bioverativ Therapeutics Inc. Use of lentiviral vectors expressing factor viii
WO2019148412A1 (en) 2018-02-01 2019-08-08 Merck Sharp & Dohme Corp. Anti-pd-1/lag3 bispecific antibodies
CN111601825B (en) 2018-02-01 2022-11-29 信达生物制药(苏州)有限公司 Fully human anti-B Cell Maturation Antigen (BCMA) single-chain antibody and application thereof
EP3746477A1 (en) 2018-02-02 2020-12-09 Bio-Techne Corporation Compounds that modulate the interaction of vista and vsig3 and methods of making and using
MX2020008289A (en) 2018-02-08 2020-09-25 Genentech Inc Bispecific antigen-binding molecules and methods of use.
AU2019218128A1 (en) 2018-02-09 2020-09-17 Genentech, Inc. Therapeutic and diagnostic methods for mast cell-mediated inflammatory diseases
US20210137930A1 (en) 2018-02-13 2021-05-13 Iovance Biotherapeutics, Inc. Expansion of tumor infiltrating lymphocytes (tils) with adenosine a2a receptor antagonists and therapeutic combinations of tils and adenosine a2a receptor antagonists
WO2019158645A1 (en) 2018-02-14 2019-08-22 Abba Therapeutics Ag Anti-human pd-l2 antibodies
AU2019226085A1 (en) 2018-02-21 2020-09-17 Genentech, Inc. Dosing for treatment with IL-22 Fc fusion proteins
US20200399376A1 (en) 2018-02-26 2020-12-24 Genentech, Inc. Dosing for treatment with anti-tigit and anti-pd-l1 antagonist antibodies
WO2019169229A1 (en) 2018-03-01 2019-09-06 Nextcure, Inc. Klrg1 binding compositions and methods of use thereof
CA3091801A1 (en) 2018-03-02 2019-09-06 Five Prime Therapeutics, Inc. B7-h4 antibodies and methods of use thereof
NL2020520B1 (en) 2018-03-02 2019-09-12 Labo Bio Medical Invest B V Multispecific binding molecules for the prevention, treatment and diagnosis of neurodegenerative disorders
AU2019236372B2 (en) 2018-03-13 2024-06-20 F. Hoffmann-La Roche Ag Therapeutic combination of 4-1 BB agonists with anti-CD20 antibodies
CN112262155A (en) 2018-03-14 2021-01-22 表面肿瘤学公司 Antibodies that bind CD39 and uses thereof
US20200040103A1 (en) 2018-03-14 2020-02-06 Genentech, Inc. Anti-klk5 antibodies and methods of use
WO2019177543A1 (en) 2018-03-15 2019-09-19 Chugai Seiyaku Kabushiki Kaisha Anti-dengue virus antibodies having cross-reactivity to zika virus and methods of use
SG11202008593PA (en) 2018-03-21 2020-10-29 Five Prime Therapeutics Inc ANTIBODIES BINDING TO VISTA AT ACIDIC pH
CA3093740A1 (en) 2018-03-22 2019-09-26 Surface Oncology, Inc. Anti-il-27 antibodies and uses thereof
CN111886256A (en) 2018-03-23 2020-11-03 百时美施贵宝公司 anti-MICA and/or MICB antibodies and uses thereof
CN112154153A (en) 2018-03-28 2020-12-29 百时美施贵宝公司 Interleukin-2/interleukin-2 receptor alpha fusion proteins and methods of use
CN111936625A (en) 2018-03-29 2020-11-13 豪夫迈·罗氏有限公司 Modulating lactogenic activity in mammalian cells
KR20200136465A (en) 2018-03-29 2020-12-07 화이자 인코포레이티드 LFA3 variants and compositions and uses thereof
TW202003567A (en) 2018-03-30 2020-01-16 大陸商南京傳奇生物科技有限公司 Single-domain antibodies against LAG-3 and uses thereof
KR20200139219A (en) 2018-04-02 2020-12-11 브리스톨-마이어스 스큅 컴퍼니 Anti-TREM-1 antibodies and uses thereof
TW202011029A (en) 2018-04-04 2020-03-16 美商建南德克公司 Methods for detecting and quantifying FGF21
WO2019193375A1 (en) 2018-04-04 2019-10-10 INSERM (Institut National de la Santé et de la Recherche Médicale) Use of fzd7 inhibitors for the treatment of retinal neovascularization
EP3552631A1 (en) 2018-04-10 2019-10-16 Inatherys Antibody-drug conjugates and their uses for the treatment of cancer
WO2019200357A1 (en) 2018-04-12 2019-10-17 Surface Oncology, Inc. Biomarker for cd47 targeting therapeutics and uses therefor
EP3773721A4 (en) 2018-04-13 2022-01-26 Genentech, Inc. Stable anti-cd79b immunoconjugate formulations
CN112585165A (en) 2018-04-25 2021-03-30 普罗米修斯生物科学公司 Optimized anti-TL 1A antibodies
WO2019207030A1 (en) 2018-04-26 2019-10-31 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for predicting a response with an immune checkpoint inhibitor in a patient suffering from a lung cancer
WO2019213384A1 (en) 2018-05-03 2019-11-07 University Of Rochester Anti-influenza neuraminidase monoclonal antibodies and uses thereof
CN110464842B (en) 2018-05-11 2022-10-14 信达生物制药(苏州)有限公司 Formulations comprising anti-PCSK 9 antibodies and uses thereof
BR112020022164A2 (en) 2018-05-18 2021-02-02 Bioverativ Therapeutics Inc. methods of treating hemophilia a
WO2019226658A1 (en) 2018-05-21 2019-11-28 Compass Therapeutics Llc Multispecific antigen-binding compositions and methods of use
CA3099308A1 (en) 2018-05-21 2019-11-28 Compass Therapeutics Llc Compositions and methods for enhancing the killing of target cells by nk cells
UY38247A (en) 2018-05-30 2019-12-31 Novartis Ag ANTIBODIES AGAINST ENTPD2, COMBINATION THERAPIES AND METHODS OF USE OF ANTIBODIES AND COMBINATION THERAPIES
JP7490574B2 (en) 2018-05-31 2024-05-27 ノバルティス アーゲー Hepatitis B antibody
SG11202010579XA (en) 2018-06-01 2020-12-30 Novartis Ag Binding molecules against bcma and uses thereof
KR20210056288A (en) 2018-06-01 2021-05-18 타유 후아시아 바이오테크 메디컬 그룹 컴퍼니 리미티드 Compositions for treating diseases or conditions and uses thereof
EP3805400A4 (en) 2018-06-04 2022-06-29 Chugai Seiyaku Kabushiki Kaisha Antigen-binding molecule showing changed half-life in cytoplasm
JP7457661B2 (en) 2018-06-04 2024-03-28 バイオジェン・エムエイ・インコーポレイテッド Anti-VLA-4 antibodies with reduced effector function
US20210155710A1 (en) 2018-06-05 2021-05-27 Amgen Inc. Modulating antibody dependent cellular phagocytosis
US12065476B2 (en) 2018-06-15 2024-08-20 Alpine Immune Sciences, Inc. PD-1 variant immunomodulatory proteins and uses thereof
JP7511308B2 (en) 2018-06-18 2024-07-05 ユーリカ セラピューティックス， インコーポレイテッド Prostate-specific membrane antigen (psma) targeting constructs and uses thereof
AU2019288728A1 (en) 2018-06-23 2021-01-14 Genentech, Inc. Methods of treating lung cancer with a pd-1 axis binding antagonist, a platinum agent, and a topoisomerase ii inhibitor
BR112020026512A2 (en) 2018-07-03 2021-04-06 Bristol-Myers Squibb Company FGF-21 FORMULATIONS
JP7411627B2 (en) 2018-07-09 2024-01-11 ファイヴ プライム セラピューティクス インク Antibody that binds to ILT4
WO2020014306A1 (en) 2018-07-10 2020-01-16 Immunogen, Inc. Met antibodies and immunoconjugates and uses thereof
CA3104536A1 (en) 2018-07-11 2020-01-16 Bristol-Myers Squibb Company Antibodies binding to vista at acidic ph
US20200171146A1 (en) 2018-07-18 2020-06-04 Genentech, Inc. Methods of treating lung cancer with a pd-1 axis binding antagonist, an antimetabolite, and a platinum agent
AU2019306628A1 (en) 2018-07-20 2021-02-11 Surface Oncology, LLC Anti-CD112R compositions and methods
US11370837B2 (en) 2018-07-25 2022-06-28 Innovent Biologics (Suzhou) Co., Ltd. Anti-TIGIT antibody and use thereof
AU2019315213B2 (en) 2018-08-01 2024-05-02 Chugai Seiyaku Kabushiki Kaisha A pharmaceutical composition for use in the treatment or prevention of a C5-related disease and a method for treating or preventing a C5-related disease
CN118459590A (en) 2018-08-01 2024-08-09 感应检查疗法公司 Anti-BTN 3A antibodies and their use in the treatment of cancer or infectious disorders
WO2020027330A1 (en) 2018-08-03 2020-02-06 中外製薬株式会社 Antigen-binding molecule containing two antigen-binding domains that are linked to each other
MA50586A (en) 2018-08-09 2020-09-16 Regeneron Pharma METHODS FOR EVALUATING THE BINDING AFFINITY OF AN ANTIBODY VARIANT TO THE NEONATAL FC RECEPTOR
MX2021001599A (en) 2018-08-09 2021-07-02 Bioverativ Therapeutics Inc Nucleic acid molecules and uses thereof for non-viral gene therapy.
US20210309746A1 (en) 2018-08-09 2021-10-07 Compass Therapeutics Llc Antibodies that bind cd277 and uses thereof
WO2020033925A2 (en) 2018-08-09 2020-02-13 Compass Therapeutics Llc Antibodies that bind cd277 and uses thereof
WO2020033923A1 (en) 2018-08-09 2020-02-13 Compass Therapeutics Llc Antigen binding agents that bind cd277 and uses thereof
WO2020033827A1 (en) 2018-08-10 2020-02-13 Genentech, Inc. Cell culture strategies for modulating protein glycosylation
SG11202003531WA (en) 2018-08-10 2020-05-28 Chugai Pharmaceutical Co Ltd Anti-cd137 antigen-binding molecule and utilization thereof
TW202021618A (en) 2018-08-17 2020-06-16 美商２３與我有限公司 Anti-il1rap antibodies and methods of use thereof
TW202031273A (en) 2018-08-31 2020-09-01 美商艾歐凡斯生物治療公司 Treatment of nsclc patients refractory for anti-pd-1 antibody
GB201814281D0 (en) 2018-09-03 2018-10-17 Femtogenix Ltd Cytotoxic agents
WO2020051507A1 (en) 2018-09-06 2020-03-12 The Broad Institute, Inc. Nucleic acid assemblies for use in targeted delivery
SG11202100888WA (en) 2018-09-07 2021-02-25 Pfizer Anti-avb8 antibodies and compositions and uses thereof
WO2020053742A2 (en) 2018-09-10 2020-03-19 Novartis Ag Anti-hla-hbv peptide antibodies
AU2019339895A1 (en) 2018-09-11 2021-03-11 Amgen Inc. Methods of modulating antibody-dependent cell-mediated cytotoxicity
US20220098613A1 (en) 2018-09-12 2022-03-31 Fred Hutchinson Cancer Research Center Reducing cd33 expression to selectively protect therapeutic cells
KR20210089146A (en) 2018-09-19 2021-07-15 알파인 이뮨 사이언시즈, 인코포레이티드 Methods and uses of variant CD80 proteins and related constructs
CA3111401A1 (en) 2018-09-19 2020-03-26 Genentech, Inc. Therapeutic and diagnostic methods for bladder cancer
JP7475336B2 (en) 2018-09-21 2024-04-26 ジェネンテック， インコーポレイテッド Diagnostic methods for triple-negative breast cancer
EP3856772A1 (en) 2018-09-25 2021-08-04 Institut National de la Santé et de la Recherche Médicale (INSERM) Use of antagonists of th17 cytokines for the treatment of bronchial remodeling in patients suffering from allergic asthma
KR20210069058A (en) 2018-09-26 2021-06-10 지앙수 헨그루이 메디슨 컴퍼니 리미티드 Anti-OX40 antibody, antigen-binding fragment thereof and pharmaceutical use
US20220002370A1 (en) 2018-09-27 2022-01-06 Xilio Development, Inc. Masked cytokine polypeptides
WO2020070062A1 (en) 2018-10-01 2020-04-09 INSERM (Institut National de la Santé et de la Recherche Médicale) Use of tim-3 inhibitors for the treatment of exacerbations in patients suffering from severe asthma
US20210340278A1 (en) 2018-10-04 2021-11-04 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods and pharmaceutical composition for the treatment of mucosal inflammatory diseases
JP2022512648A (en) 2018-10-09 2022-02-07 アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル Use of αV-integrin (CD51) inhibitors for the treatment of myocardial fibrosis
CN113164780A (en) 2018-10-10 2021-07-23 泰洛斯治疗公司 anti-LAP antibody variants and uses thereof
UY38407A (en) 2018-10-15 2020-05-29 Novartis Ag TREM2 STABILIZING ANTIBODIES
WO2020081493A1 (en) 2018-10-16 2020-04-23 Molecular Templates, Inc. Pd-l1 binding proteins
WO2020081767A1 (en) 2018-10-18 2020-04-23 Genentech, Inc. Diagnostic and therapeutic methods for sarcomatoid kidney cancer
WO2020086758A1 (en) 2018-10-23 2020-04-30 Dragonfly Therapeutics, Inc. Heterodimeric fc-fused proteins
EP3873532A1 (en) 2018-10-31 2021-09-08 Novartis AG Dc-sign antibody drug conjugates
WO2020089437A1 (en) 2018-10-31 2020-05-07 Engmab Sàrl Combination therapy
US20210395390A1 (en) 2018-10-31 2021-12-23 Bayer Aktiengesellschaft Reversal agents for neutralizing the therapeutic activity of anti-fxia antibodies
TW202031899A (en) 2018-11-05 2020-09-01 美商建南德克公司 Methods of producing two chain proteins in prokaryotic host cells
TW202039831A (en) 2018-11-05 2020-11-01 美商艾歐凡斯生物治療公司 Treatment of nsclc patients refractory for anti-pd-1 antibody
BR112021009373A2 (en) 2018-11-16 2021-08-17 Memorial Sloan Kettering Cancer Center antibodies to mucin-16 and methods of using them
TWI779253B (en) 2018-11-27 2022-10-01 大陸商信達生物製藥（蘇州）有限公司 ANTI-IL-23p19 ANTIBODY AND USE THEREOF
JP2022513653A (en) 2018-11-28 2022-02-09 ブリストル－マイヤーズ スクイブ カンパニー Antibodies containing modified heavy chain constant regions
KR20210135987A (en) 2018-11-30 2021-11-16 알파인 이뮨 사이언시즈, 인코포레이티드 CD86 variant immunomodulatory protein and uses thereof
JP2022511502A (en) 2018-12-05 2022-01-31 ジェネンテック， インコーポレイテッド Diagnostic Methods and Diagnostic Compositions for Cancer Immunotherapy
US20220041719A1 (en) 2018-12-05 2022-02-10 Morphosys Ag Multispecific antigen-binding molecules
WO2020117257A1 (en) 2018-12-06 2020-06-11 Genentech, Inc. Combination therapy of diffuse large b-cell lymphoma comprising an anti-cd79b immunoconjugates, an alkylating agent and an anti-cd20 antibody
US20220098310A1 (en) 2018-12-06 2022-03-31 Alexion Pharmaceuticals, Inc. Anti-alk2 antibodies and uses thereof
JP2022513198A (en) 2018-12-10 2022-02-07 ジェネンテック， インコーポレイテッド Photocrosslinkable peptide for site-specific conjugation to Fc-containing proteins
WO2020127509A1 (en) 2018-12-18 2020-06-25 Catapult Therapeutics B.V. The use of anti-ccr7 mabs for the prevention or treatment of graft-versus-host disease (gvhd)
TW202039554A (en) 2018-12-19 2020-11-01 瑞士商諾華公司 Anti-tnf-alpha antibodies
AR117453A1 (en) 2018-12-20 2021-08-04 Genentech Inc CF OF MODIFIED ANTIBODIES AND METHODS TO USE THEM
AR117327A1 (en) 2018-12-20 2021-07-28 23Andme Inc ANTI-CD96 ANTIBODIES AND METHODS OF USE OF THEM
WO2020132231A1 (en) 2018-12-21 2020-06-25 Genentech, Inc. Methods of producing polypeptides using a cell line resistant to apoptosis
CN113195531A (en) 2018-12-21 2021-07-30 豪夫迈·罗氏有限公司 Antibodies that bind to VEGF and IL-1beta and methods of use thereof
EA202191656A1 (en) 2018-12-21 2021-10-26 23Эндми, Инк. ANTI-IL-36 ANTIBODIES AND METHODS OF THEIR APPLICATION
PE20211296A1 (en) 2018-12-21 2021-07-20 Novartis Ag ANTI-PMEL17 ANTIBODIES AND CONJUGATES THEREOF
CN111349162A (en) 2018-12-21 2020-06-30 神州细胞工程有限公司 Humanized anti-PD-1 antibodies and uses thereof
AU2019416216A1 (en) 2018-12-26 2021-07-08 City Of Hope Activatable masked anti-CTLA4 binding proteins
BR112021011642A2 (en) * 2018-12-27 2021-11-03 Kindred Biosciences Inc igg fc variants for veterinary use
CN113272327A (en) 2018-12-30 2021-08-17 豪夫迈·罗氏有限公司 Anti-rabbit CD19 antibodies and methods of use thereof
BR112021013157A8 (en) 2019-01-03 2022-12-06 Inst Nat Sante Rech Med USES OF AN NRP-1 INHIBITOR, USE OF A COMBINATION, USE OF A MULTIESPECIFIC ANTIBODY, EX VIVO METHOD TO PREDICT, USE OF AN INHIBITOR, MULTIESPECIFIC ANTIBODY, MODIFIED CELL POPULATION, EX VIVO METHOD OF PRODUCTION AND USE OF A POPULATION OF T CELLS
SG11202106686PA (en) 2019-01-04 2021-07-29 Resolve Therapeutics Llc Treatment of sjogren's disease with nuclease fusion proteins
CA3124837A1 (en) 2019-01-14 2020-07-23 Genentech, Inc. Methods of treating cancer with a pd-1 axis binding antagonist and an rna vaccine
WO2020148207A1 (en) 2019-01-14 2020-07-23 INSERM (Institut National de la Santé et de la Recherche Médicale) Human monoclonal antibodies binding to hla-a2
JP7539897B2 (en) 2019-01-16 2024-08-26 コンパス セラピューティクス リミテッド ライアビリティ カンパニー Preparations of antibodies that bind to human CD137 and uses thereof
CA3127236A1 (en) 2019-01-22 2020-07-30 Bristol-Myers Squibb Company Antibodies against il-7r alpha subunit and uses thereof
BR112021014276A2 (en) 2019-01-22 2021-09-28 Genentech, Inc. ISOLATED IGA ANTIBODIES, ISOLATED IGG-IGA FUSION MOLECULES, ISOLATED NUCLEIC ACID, HOST CELL, METHOD FOR PRODUCING AN ANTIBODY, TO TREAT AN INDIVIDUAL, TO INCREASE THE EXPRESSION OF DIMERS, TRIMERS OR TETRAMERS, TO INCREASE THE PRODUCTION OF POLYMERS, TO INCREASE The production of dimers, to increase the production of a polymer, to decrease the production of polymers, to increase the transient expression of an antibody, to express dimers of fusion molecules, to express dimers, trimers or tetramers, to purify an antibody, TO PURIFY AN OLIOMER STATE OF AN ANTIBODY, PHARMACEUTICAL COMPOSITION AND USE OF THE ANTIBODY
JP2022523475A (en) 2019-01-23 2022-04-25 ジェネンテック， インコーポレイテッド How to produce multimeric proteins in eukaryotic host cells
WO2020154595A1 (en) 2019-01-24 2020-07-30 Massachusetts Institute Of Technology Nucleic acid nanostructure platform for antigen presentation and vaccine formulations formed therefrom
US20220089770A1 (en) 2019-01-24 2022-03-24 Chugai Seiyaku Kabushiki Kaisha Novel cancer antigens and antibodies of said antigens
GB201901197D0 (en) 2019-01-29 2019-03-20 Femtogenix Ltd G-A Crosslinking cytotoxic agents
EP3931220A1 (en) 2019-02-27 2022-01-05 F. Hoffmann-La Roche AG Dosing for treatment with anti-tigit and anti-cd20 or anti-cd38 antibodies
JP2022522473A (en) 2019-03-01 2022-04-19 アイオバンス バイオセラピューティクス，インコーポレイテッド Expansion culture of tumor-infiltrating lymphocytes from humoral tumors and their therapeutic use
CA3126728A1 (en) 2019-03-08 2020-09-17 Genentech, Inc. Methods for detecting and quantifying membrane-associated proteins on extracellular vesicles
US20220135682A1 (en) 2019-03-11 2022-05-05 Jounce Therapeutics, Inc. Anti-ICOS Antibodies for the Treatment of Cancer
SG11202109424RA (en) 2019-03-14 2021-09-29 Genentech Inc Treatment of cancer with her2xcd3 bispecific antibodies in combination with anti-her2 mab
JP2022525937A (en) 2019-03-20 2022-05-20 イムチェック セラピューティクス エスエーエス Antibodies with specificity for BTN2 and their use
JP2022527345A (en) 2019-04-03 2022-06-01 オレガ・バイオテック Combined therapy based on PD1 and IL-17B inhibitors
EP3947458A1 (en) 2019-04-03 2022-02-09 Genzyme Corporation Anti-alpha beta tcr binding polypeptides with reduced fragmentation
EA202192400A1 (en) 2019-04-08 2022-01-25 Биоген Ма Инк. ANTIBODIES AGAINST INTEGRIN AND THEIR USE
GB2589049C (en) 2019-04-11 2024-02-21 argenx BV Anti-IgE antibodies
WO2020214690A1 (en) 2019-04-15 2020-10-22 Qwixel Therapeutics Fusion protein composition(s) comprising targeted masked type i interferons (ifna and ifnb) and an antibody against tumor antigen, for use in the treatment of cancer
WO2020214867A1 (en) 2019-04-17 2020-10-22 Alpine Immune Sciences, Inc. Methods and uses of variant icos ligand (icosl) fusion proteins
TW202043291A (en) 2019-04-19 2020-12-01 美商建南德克公司 Anti-mertk antibodies and methods of use
EP3962947A2 (en) 2019-05-03 2022-03-09 F. Hoffmann-La Roche AG Methods of treating cancer with an anti-pd-l1 antibody
TW202108178A (en) 2019-05-14 2021-03-01 美商建南德克公司 METHODS OF USING ANTI-CD79b IMMUNOCONJUGATES TO TREAT FOLLICULAR LYMPHOMA
EP3972997A1 (en) 2019-05-20 2022-03-30 Institut National de la Santé et de la Recherche Médicale (INSERM) Novel anti-cd25 antibodies
KR20220010527A (en) 2019-05-20 2022-01-25 노파르티스 아게 Antibody drug conjugates having a linker comprising a hydrophilic group
EP3972649A2 (en) 2019-05-20 2022-03-30 Novartis AG Mcl-1 inhibitor antibody-drug conjugates and methods of use
JOP20210309A1 (en) 2019-05-21 2023-01-30 Novartis Ag Cd19 binding molecules and uses thereof
EP3972634A2 (en) 2019-05-21 2022-03-30 Novartis AG Trispecific binding molecules against bcma and uses thereof
WO2020236797A1 (en) 2019-05-21 2020-11-26 Novartis Ag Variant cd58 domains and uses thereof
EP3972692A1 (en) 2019-05-23 2022-03-30 AC Immune SA Anti-tdp-43 binding molecules and uses thereof
KR20220019018A (en) 2019-06-07 2022-02-15 아르제넥스 비브이비에이 Pharmaceutical formulations of FcRn inhibitors suitable for subcutaneous administration
MX2021015007A (en) 2019-06-10 2022-01-31 Chugai Pharmaceutical Co Ltd Anti-t cell antigen-binding molecule to be used in combination with cytokine inhibitor.
WO2020250159A1 (en) 2019-06-12 2020-12-17 Novartis Ag Natriuretic peptide receptor 1 antibodies and methods of use
CA3143584A1 (en) 2019-06-18 2020-12-24 Bayer Aktiengesellschaft Adrenomedullin-analogues for long-term stabilization and their use
US11905532B2 (en) 2019-06-25 2024-02-20 Massachusetts Institute Of Technology Compositions and methods for molecular memory storage and retrieval
BR112021023173A2 (en) 2019-07-10 2022-01-04 Chugai Pharmaceutical Co Ltd Claudin-6 binding molecules and uses thereof
WO2021005232A1 (en) 2019-07-11 2021-01-14 Umc Utrecht Holding B.V. Intranasal administration of neutralising antiviral antibodies
AR119382A1 (en) 2019-07-12 2021-12-15 Hoffmann La Roche PRE-TARGETING ANTIBODIES AND METHODS OF USE
JPWO2021010326A1 (en) 2019-07-12 2021-01-21
CN114144435B (en) 2019-07-15 2024-06-25 百时美施贵宝公司 Antibodies against human TREM-1 and uses thereof
CN114174536A (en) 2019-07-15 2022-03-11 百时美施贵宝公司 anti-TREM-1 antibodies and uses thereof
KR20220034807A (en) 2019-07-16 2022-03-18 인쎄름 (엥스띠뛰 나씨오날 드 라 쌍떼 에 드 라 흐쉐르슈 메디깔) Antibodies having specificity for CD38 and uses thereof
BR112022001017A2 (en) 2019-07-19 2022-04-12 Sinocelltech Ltd Humanized anti-vegf monoclonal antibody
WO2021013065A1 (en) 2019-07-19 2021-01-28 神州细胞工程有限公司 Humanized anti-vegf fab antibody fragment and use thereof
US20220306734A1 (en) 2019-07-24 2022-09-29 H. Lundbeck A/S Anti-mglur5 antibodies and uses thereof
WO2021016598A1 (en) 2019-07-25 2021-01-28 Immunowake Inc. Methods of measuring cell-mediated killing by effectors
WO2021018035A1 (en) 2019-07-26 2021-02-04 神州细胞工程有限公司 Humanized anti-il17a antibody and use thereof
CN112300279A (en) 2019-07-26 2021-02-02 上海复宏汉霖生物技术股份有限公司 Methods and compositions directed to anti-CD 73 antibodies and variants
JP2022542505A (en) 2019-07-29 2022-10-04 コンピュジェン リミテッド ANTI-PVRIG ANTIBODY FORMULATIONS AND THEIR USE
AU2020322165A1 (en) 2019-07-31 2022-01-06 F. Hoffmann-La Roche Ag Dosage and administration regimen for the treatment or prevention of C5-related diseases by the use of the anti-C5 antibody Crovalimab
WO2021019033A1 (en) 2019-07-31 2021-02-04 F. Hoffmann-La Roche Ag Dosage and administration regimen for the treatment or prevention of c5-related diseases by the use of the anti-c5 antibody crovalimab
CN114641490B (en) 2019-08-06 2023-06-06 新旭生技股份有限公司 Antibodies that bind to pathological TAU species and uses thereof
CN114867751A (en) 2019-08-12 2022-08-05 阿帕特夫研究和发展有限公司 4-1BB and OX40 binding proteins and related compositions and methods, anti-4-1 BB antibodies, anti-OX 40 antibodies
TW202122420A (en) 2019-08-30 2021-06-16 美商艾吉納斯公司 Anti-cd96 antibodies and methods of use thereof
JP2022547071A (en) 2019-09-04 2022-11-10 インサーム（インスティテュ ナシオナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシェ メディカル） Antibodies specific to IL20-RB and their use for the treatment of acute exacerbations of chronic obstructive pulmonary disease
WO2021050645A1 (en) 2019-09-12 2021-03-18 Genentech, Inc. Compositions and methods of treating lupus nephritis
WO2021055329A1 (en) 2019-09-16 2021-03-25 Surface Oncology, Inc. Anti-cd39 antibody compositions and methods
US11918649B2 (en) 2019-09-18 2024-03-05 Molecular Templates, Inc. PD-L1-binding molecules comprising Shiga toxin a subunit scaffolds
TW202124446A (en) 2019-09-18 2021-07-01 瑞士商諾華公司 Combination therapies with entpd2 antibodies
PE20221906A1 (en) 2019-09-18 2022-12-23 Genentech Inc ANTI-KLK7 ANTIBODIES, ANTI-KLK5 ANTIBODIES, ANTI-KLK5/KLK7 MULTISPECIFIC ANTIBODIES AND METHODS OF USE
JP2022548881A (en) 2019-09-18 2022-11-22 ノバルティス アーゲー ENTPD2 Antibodies, Combination Therapy and Methods of Using Antibodies and Combination Therapy
US20230192860A1 (en) 2019-09-19 2023-06-22 Bristol-Myers Squibb Company Antibodies Binding to Vista at Acidic pH
KR20220066295A (en) 2019-09-20 2022-05-24 제넨테크, 인크. Dosing of Anti-Tryptase Antibodies
CA3151078A1 (en) 2019-09-25 2021-04-01 Surface Oncology, Inc. Anti-il-27 antibodies and uses thereof
US20220411511A1 (en) 2019-09-26 2022-12-29 Stcube & Co. Antibodies specific to glycosylated ctla-4 and methods of use thereof
JP2022550067A (en) 2019-09-27 2022-11-30 ヤンセン バイオテツク，インコーポレーテツド Anti-CEACAM antibody and use thereof
AU2020351734A1 (en) 2019-09-27 2022-04-14 Genentech, Inc. Dosing for treatment with anti-TIGIT and anti-PD-L1 antagonist antibodies
JP2022549932A (en) 2019-09-30 2022-11-29 バイオベラティブ セラピューティクス インコーポレイテッド Lentiviral vector preparation
EP3800201A1 (en) 2019-10-01 2021-04-07 INSERM (Institut National de la Santé et de la Recherche Médicale) Cd28h stimulation enhances nk cell killing activities
JP2022550434A (en) 2019-10-04 2022-12-01 ティーエーイー ライフ サイエンシーズ Antibody compositions containing Fc mutations and site-specific conjugation properties
JP2022552282A (en) 2019-10-09 2022-12-15 エスティーキューブ アンド カンパニー Antibodies specific for glycosylated LAG3 and methods of use thereof
BR112022007216A2 (en) 2019-10-18 2022-08-23 Genentech Inc METHODS FOR TREATMENT OF DIFFUSE LYMPHOMA, KIT AND IMMUNOCONJUGATE
WO2021080682A1 (en) 2019-10-24 2021-04-29 Massachusetts Institute Of Technology Monoclonal antibodies that bind human cd161 and uses thereof
BR112022007720A2 (en) 2019-10-24 2022-08-23 Prometheus Biosciences Inc HUMANIZED ANTIBODIES TO TNF-LIKE BINDING 1A (TL1A) AND THEIR USES
US20230002785A1 (en) 2019-10-28 2023-01-05 Georgia Tech Research Corporation Mrna-encoded antibodies for contraception
US20240148901A1 (en) 2019-10-28 2024-05-09 Georgia Tech Research Corporation Compositions and methods for prophylaxis of hiv
JP2023500651A (en) 2019-11-04 2023-01-10 コンピュジェン リミテッド Combination therapy with anti-PVRIG antibody formulation and anti-PD-1 antibody
JP2022554270A (en) 2019-11-05 2022-12-28 ジョンス セラピューティクス， インコーポレイテッド Methods of treating cancer with anti-PD-1 antibodies
MX2022005400A (en) 2019-11-06 2022-05-24 Genentech Inc Diagnostic and therapeutic methods for treatment of hematologic cancers.
US20230040928A1 (en) 2019-12-09 2023-02-09 INSERM (Institut National de la Santé et de la Recherche Médicale) Antibodies having specificity to her4 and uses thereof
AR120741A1 (en) 2019-12-13 2022-03-16 Genentech Inc ANTI-LY6G6D ANTIBODIES AND METHODS OF USE
WO2021123173A1 (en) 2019-12-20 2021-06-24 F. Hoffmann-La Roche Ag Il-37 fusion proteins and uses thereof
KR102645629B1 (en) 2019-12-27 2024-03-07 추가이 세이야쿠 가부시키가이샤 Anti-CTLA-4 antibodies and their uses
CN113045655A (en) 2019-12-27 2021-06-29 高诚生物医药(香港)有限公司 anti-OX 40 antibodies and uses thereof
CA3160204A1 (en) 2020-01-06 2021-07-15 Vaccinex, Inc. Anti-ccr8 antibodies and uses thereof
US20210236596A1 (en) 2020-01-08 2021-08-05 argenx BV Methods for treating pemphigus disorders
CN110818795B (en) 2020-01-10 2020-04-24 上海复宏汉霖生物技术股份有限公司 anti-TIGIT antibodies and methods of use
CN115190814A (en) 2020-01-24 2022-10-14 辉瑞公司 anti-E-selectin antibodies, compositions, and methods of use
WO2022050954A1 (en) 2020-09-04 2022-03-10 Genentech, Inc. Dosing for treatment with anti-tigit and anti-pd-l1 antagonist antibodies
WO2021194481A1 (en) 2020-03-24 2021-09-30 Genentech, Inc. Dosing for treatment with anti-tigit and anti-pd-l1 antagonist antibodies
EP4097143A1 (en) 2020-01-31 2022-12-07 The Cleveland Clinic Foundation Anti-müllerian hormone receptor 2 antibodies and methods of use
IL294859A (en) 2020-01-31 2022-09-01 Genentech Inc Methods of inducing neoepitope-specific t cells with a pd-1 axis binding antagonist and an rna vaccine
EP4100426A1 (en) 2020-02-06 2022-12-14 Bristol-Myers Squibb Company Il-10 and uses thereof
TW202144395A (en) 2020-02-12 2021-12-01 日商中外製藥股份有限公司 Anti-CD137 antigen-binding molecule for use in cancer treatment
WO2021163064A2 (en) 2020-02-14 2021-08-19 Jounce Therapeutics, Inc. Antibodies and fusion proteins that bind to ccr8 and uses thereof
BR112022016491A2 (en) 2020-02-28 2022-10-11 Shanghai Henlius Biotech Inc ANTI-CD137 CONSTRUCTION AND USES THEREOF
CA3169521A1 (en) 2020-02-28 2021-09-02 Marie-Priscille Brun Modified binding polypeptides for optimized drug conjugation
WO2021170750A1 (en) 2020-02-28 2021-09-02 Orega Biotech Combination therapies based on ctla4 and il-17b inhibitors
AU2021228078A1 (en) 2020-02-28 2022-09-22 Shanghai Henlius Biotech, Inc. Anti-CD137 constructs, multispecific antibody and uses thereof
WO2021175954A1 (en) 2020-03-04 2021-09-10 Imcheck Therapeutics Sas Antibodies having specificity for btnl8 and uses thereof
US11896619B2 (en) 2020-03-10 2024-02-13 Massachusetts Institute Of Technology Compositions and methods for immunotherapy of NPM1c-positive cancer
KR20220152262A (en) 2020-03-13 2022-11-15 제넨테크, 인크. Anti-interleukin-33 antibodies and uses thereof
TW202144408A (en) 2020-03-19 2021-12-01 美商建南德克公司 Isoform-selective anti-tgf-beta antibodies and methods of use
US11525137B2 (en) 2020-03-19 2022-12-13 Avidity Biosciences, Inc. Compositions and methods of treating Facioscapulohumeral muscular dystrophy
US11365239B2 (en) 2020-03-20 2022-06-21 Tsb Therapeutics (Beijing) Co., Ltd. Anti-SARS-COV-2 antibodies and uses thereof
CN115867649A (en) 2020-03-24 2023-03-28 基因泰克公司 Tie 2-binding agents and methods of use
CN111122879B (en) * 2020-03-26 2020-07-14 珠海丽珠试剂股份有限公司 Coating liquid of novel coronavirus recombinant antigen, pretreatment method, application and product
EP4127153A2 (en) 2020-03-26 2023-02-08 Genentech, Inc. Modified mammalian cells having reduced host cell proteins
JP2023520773A (en) 2020-03-27 2023-05-19 ノバルティス アーゲー Bispecific combination therapy for treating proliferative and autoimmune diseases
WO2021198034A1 (en) 2020-03-30 2021-10-07 F. Hoffmann-La Roche Ag Antibody that binds to vegf and pdgf-b and methods of use
WO2021202235A1 (en) 2020-04-01 2021-10-07 University Of Rochester Monoclonal antibodies against the hemagglutinin (ha) and neuraminidase (na) of influenza h3n2 viruses
JP2023520515A (en) 2020-04-03 2023-05-17 ジェネンテック， インコーポレイテッド Therapeutic and diagnostic methods for cancer
EP4132954A1 (en) 2020-04-08 2023-02-15 Institut National de la Santé et de la Recherche Médicale (INSERM) Use of cdon inhibitors for the treatment of endothelial dysfunction
US20230272056A1 (en) 2020-04-09 2023-08-31 Merck Sharp & Dohme Llc Affinity matured anti-lap antibodies and uses thereof
WO2021204179A1 (en) 2020-04-09 2021-10-14 Suzhou Abogen Biosciences Co., Ltd. Nucleic acid vaccines for coronavirus
WO2021207662A1 (en) 2020-04-10 2021-10-14 Genentech, Inc. Use of il-22fc for the treatment or prevention of pneumonia, acute respiratory distress syndrome, or cytokine release syndrome
CA3173134A1 (en) 2020-04-20 2021-10-28 Jounce Therapeutics, Inc. Compositions and methods for vaccination and the treatment of infectious diseases
BR112022021203A2 (en) 2020-04-24 2022-12-06 Hoffmann La Roche ENZYME MODULATION AND PATHWAY WITH SULPHYDRYL COMPOUNDS AND THEIR DERIVATIVES
EP4139359A1 (en) 2020-04-24 2023-03-01 Genentech, Inc. Methods of using anti-cd79b immunoconjugates
WO2021222167A1 (en) 2020-04-28 2021-11-04 Genentech, Inc. Methods and compositions for non-small cell lung cancer immunotherapy
JP2023523480A (en) 2020-04-28 2023-06-06 ザ ロックフェラー ユニバーシティー Neutralizing anti-SARS-COV-2 antibodies and methods of use thereof
AU2021265205A1 (en) 2020-04-30 2023-01-05 Academisch Ziekenhuis Groningen Anti-CD103 antibodies
US20230181756A1 (en) 2020-04-30 2023-06-15 Novartis Ag Ccr7 antibody drug conjugates for treating cancer
WO2021226553A2 (en) 2020-05-08 2021-11-11 Alpine Immune Sciences, Inc. April and baff inhibitory immunomodulatory proteins with and without a t cell inhibitory protein and methods of use thereof
JP2023525053A (en) 2020-05-12 2023-06-14 インサーム（インスティテュ ナシオナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシェ メディカル） A new method to treat cutaneous T-cell lymphoma and TFH-derived lymphoma
US20230192867A1 (en) 2020-05-15 2023-06-22 Bristol-Myers Squibb Company Antibodies to garp
BR112022023088A2 (en) 2020-05-17 2022-12-20 Astrazeneca Uk Ltd ANTIBODIES AGAINST SARS-COV-2 AND SELECTION METHODS AND THEIR USE
GB2595299B (en) 2020-05-21 2022-08-03 Mabsolve Ltd Modified immunoglobulin FC regions
EP4157881A1 (en) 2020-05-27 2023-04-05 Staidson (Beijing) Biopharmaceuticals Co., Ltd. Antibodies specifically recognizing nerve growth factor and uses thereof
CR20220608A (en) 2020-05-29 2023-01-26 23Andme Inc Anti-cd200r1 antibodies and methods of use thereof
MX2022015376A (en) 2020-06-02 2023-04-14 Dynamicure Biotechnology Llc Anti-cd93 constructs and uses thereof.
CN116529260A (en) 2020-06-02 2023-08-01 当康生物技术有限责任公司 anti-CD 93 constructs and uses thereof
CA3184495A1 (en) 2020-06-08 2021-12-16 F. Hoffmann-La Roche Ag Anti-hbv antibodies and methods of use
JP2023529206A (en) 2020-06-12 2023-07-07 ジェネンテック， インコーポレイテッド Methods and compositions for cancer immunotherapy
MX2022015877A (en) 2020-06-16 2023-01-24 Genentech Inc Methods and compositions for treating triple-negative breast cancer.
AU2021293507A1 (en) 2020-06-18 2023-02-02 F. Hoffmann-La Roche Ag Treatment with anti-TIGIT antibodies and PD-1 axis binding antagonists
PE20230435A1 (en) 2020-06-19 2023-03-08 Chugai Pharmaceutical Co Ltd ANTI-T CELL ANTIGEN BINDING MOLECULES TO BE USED IN COMBINATION WITH AN ANGIOGENESIS INHIBITOR
CA3190173A1 (en) 2020-06-22 2021-12-30 Innovent Biologics (Suzhou) Co., Ltd. Anti-ang-2 antibody and use thereof
US20230235040A1 (en) 2020-06-22 2023-07-27 Almirall, S.A. Anti-il-36 antibodies and methods of use thereof
EP4169948A4 (en) 2020-06-22 2024-07-10 Innovent Biologics Suzhou Co Ltd Anti-cd73 antibody and use thereof
IL299161A (en) 2020-06-24 2023-02-01 Genentech Inc Apoptosis resistant cell lines
AU2021297873A1 (en) 2020-06-25 2023-02-09 Merck Sharp & Dohme Llc High affinity antibodies targeting tau phosphorylated at serine 413
CA3165342A1 (en) 2020-06-29 2022-01-06 James Arthur Posada Treatment of sjogren's syndrome with nuclease fusion proteins
MX2023000339A (en) 2020-07-10 2023-02-09 Hoffmann La Roche Antibodies which bind to cancer cells and target radionuclides to said cells.
MX2023000617A (en) 2020-07-17 2023-02-13 Genentech Inc Anti-notch2 antibodies and methods of use.
GB2597532A (en) 2020-07-28 2022-02-02 Femtogenix Ltd Cytotoxic compounds
WO2022026763A1 (en) 2020-07-29 2022-02-03 Dynamicure Biotechnology Llc Anti-cd93 constructs and uses thereof
EP4189121A1 (en) 2020-08-03 2023-06-07 Genentech, Inc. Diagnostic and therapeutic methods for lymphoma
WO2022031978A1 (en) 2020-08-06 2022-02-10 Bioverativ Usa Inc. Inflammatory cytokines and fatigue in subject with a complement mediated disease
KR20230045613A (en) 2020-08-10 2023-04-04 아스트라제네카 유케이 리미티드 SARS-COV-2 Antibodies for Treatment and Prevention of COVID-19
EP4204448A2 (en) 2020-08-27 2023-07-05 cureab GmbH Anti-golph2 antibodies for macrophage and dendritic cell differentiation
TW202227625A (en) 2020-08-28 2022-07-16 美商建南德克公司 Crispr/cas9 multiplex knockout of host cell proteins
CN116323663A (en) 2020-09-04 2023-06-23 豪夫迈·罗氏有限公司 Antibodies that bind VEGF-A and ANG2 and methods of use thereof
JP2023541627A (en) 2020-09-14 2023-10-03 イシュノス サイエンシズ ソシエテ アノニム Antibodies that bind to IL1RAP and uses thereof
WO2022076462A1 (en) 2020-10-05 2022-04-14 Genentech, Inc. Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies
EP4225330A1 (en) 2020-10-06 2023-08-16 Iovance Biotherapeutics, Inc. Treatment of nsclc patients with tumor infiltrating lymphocyte therapies
WO2022076606A1 (en) 2020-10-06 2022-04-14 Iovance Biotherapeutics, Inc. Treatment of nsclc patients with tumor infiltrating lymphocyte therapies
WO2022081765A1 (en) 2020-10-14 2022-04-21 Denali Therapeutics Inc. Fusion proteins comprising sulfoglucosamine sulfohydrolase enzymes and methods thereof
TW202233671A (en) 2020-10-20 2022-09-01 美商建南德克公司 Peg-conjugated anti-mertk antibodies and methods of use
US20230400467A1 (en) 2020-10-26 2023-12-14 Compugen Ltd. Pvrl2 and/or pvrig as biomarkers for treatment
WO2022093981A1 (en) 2020-10-28 2022-05-05 Genentech, Inc. Combination therapy comprising ptpn22 inhibitors and pd-l1 binding antagonists
AU2021374590A1 (en) 2020-11-04 2023-06-01 Genentech, Inc. Subcutaneous dosing of anti-cd20/anti-cd3 bispecific antibodies
CA3196191A1 (en) 2020-11-04 2022-05-12 Chi-Chung Li Dosing for treatment with anti-cd20/anti-cd3 bispecific antibodies and anti-cd79b antibody drug conjugates
EP4240758A1 (en) 2020-11-04 2023-09-13 The Rockefeller University Neutralizing anti-sars-cov-2 antibodies
CA3196539A1 (en) 2020-11-04 2022-05-12 Chi-Chung Li Dosing for treatment with anti-cd20/anti-cd3 bispecific antibodies
WO2022096547A1 (en) 2020-11-05 2022-05-12 INSERM (Institut National de la Santé et de la Recherche Médicale) Use of il-6 inhibitors for the treatment of acute chest syndrome in patients suffering from sickle cell disease
IL302412A (en) 2020-11-06 2023-06-01 Novartis Ag Anti-cd19 agent and b cell targeting agent combination therapy for treating b cell malignancies
CN116472288A (en) 2020-11-06 2023-07-21 诺华股份有限公司 Antibody Fc variants
MX2023005353A (en) 2020-11-06 2023-05-22 Novartis Ag Cd19 binding molecules and uses thereof.
TW202229322A (en) 2020-11-19 2022-08-01 大陸商舒泰神（北京）生物製藥股份有限公司 Long-acting nerve growth factor polypeptides and uses thereof
CN116917318A (en) 2020-11-20 2023-10-20 法国国家健康和医学研究院 anti-CD 25 antibodies
EP4247496A1 (en) 2020-11-20 2023-09-27 Institut National de la Santé et de la Recherche Médicale (INSERM) Anti-cd25 antibodies
CA3202759A1 (en) 2020-11-24 2022-06-02 Novartis Ag Mcl-1 inhibitor antibody-drug conjugates and methods of use
WO2022112942A2 (en) 2020-11-24 2022-06-02 Novartis Ag Anti-cd48 antibodies, antibody drug conjugates, and uses thereof
AU2021387970A1 (en) 2020-11-27 2023-06-29 General Nanotherapeutics Llc Methods and composition for treatment of immune-mediated diseases
US20240301086A1 (en) 2020-12-01 2024-09-12 Aptevo Research And Development Llc Tumor-associated antigens and cd3-binding proteins, related compositions, and methods
TW202241468A (en) 2020-12-11 2022-11-01 美商艾歐凡斯生物治療公司 Treatment of cancer patients with tumor infiltrating lymphocyte therapies in combination with braf inhibitors and/or mek inhibitors
WO2022130182A1 (en) 2020-12-14 2022-06-23 Novartis Ag Reversal binding agents for anti-natriuretic peptide receptor 1 (npr1) antibodies and uses thereof
WO2022133149A1 (en) 2020-12-17 2022-06-23 Iovance Biotherapeutics, Inc. Treatment of cancers with tumor infiltrating lymphocytes
US20240299539A1 (en) 2020-12-17 2024-09-12 Iovance Biotherapeutics, Inc. Treatment with tumor infiltrating lymphocyte therapies in combination with ctla-4 and pd-1 inhibitors
WO2022140797A1 (en) 2020-12-23 2022-06-30 Immunowake Inc. Immunocytokines and uses thereof
WO2022135467A1 (en) 2020-12-23 2022-06-30 信达生物制药(苏州)有限公司 Anti-b7-h3 antibody and uses thereof
US20240110152A1 (en) 2020-12-31 2024-04-04 Iovance Biotherapeutics, Inc. Devices and processes for automated production of tumor infiltrating lymphocytes
EP4277705A1 (en) 2021-01-12 2023-11-22 F. Hoffmann-La Roche AG Split antibodies which bind to cancer cells and target radionuclides to said cells
WO2022153212A1 (en) 2021-01-13 2022-07-21 Axon Neuroscience Se Antibodies neutralizing sars-cov-2
KR20230131205A (en) 2021-01-13 2023-09-12 에프. 호프만-라 로슈 아게 combination therapy
EP4277926A1 (en) 2021-01-15 2023-11-22 The Rockefeller University Neutralizing anti-sars-cov-2 antibodies
WO2022165266A1 (en) 2021-01-28 2022-08-04 Compugen Ltd. Anti-pvrig antibodies formulations and uses thereof
US20240076373A1 (en) 2021-01-28 2024-03-07 Compugen Ltd. Combination therapy with anti-pvrig antibodies formulations and anti-pd-1 antibodies
US20240307437A1 (en) 2021-01-29 2024-09-19 Iovance Biotherapeutics, Inc. Cytokine associated tumor infiltrating lymphocytes compositions and methods
AU2022218137A1 (en) 2021-02-03 2023-08-24 Mozart Therapeutics, Inc. Binding agents and methods of using the same
JP2024509695A (en) 2021-02-03 2024-03-05 ジェネンテック， インコーポレイテッド Multispecific binding proteolysis platform and methods of use
JP2024505987A (en) 2021-02-04 2024-02-08 イノベント バイオロジクス（スーチョウ）カンパニー，リミティド Anti-TNFR2 antibodies and their uses
KR20230157356A (en) 2021-02-17 2023-11-16 프로메테우스 바이오사이언시즈, 인크. Anti-CD30L antibodies and uses thereof
WO2022187270A1 (en) 2021-03-01 2022-09-09 Xilio Development, Inc. Combination of ctla4 and pd1/pdl1 antibodies for treating cancer
TW202246324A (en) 2021-03-01 2022-12-01 美商艾希利歐發展股份有限公司 Combination of masked ctla4 and pd1/pdl1 antibodies for treating cancer
JP2024509191A (en) 2021-03-05 2024-02-29 ダイナミキュア バイオテクノロジー エルエルシー Anti-VISTA constructs and their uses
TW202300014A (en) 2021-03-05 2023-01-01 美商艾歐凡斯生物治療公司 Tumor storage and cell culture compositions
KR20230148226A (en) 2021-03-10 2023-10-24 이뮤노웨이크 인크. Immunomodulatory Molecules and Their Uses
AR125074A1 (en) 2021-03-12 2023-06-07 Genentech Inc ANTI-KLK7 ANTIBODIES, ANTI-KLK5 ANTIBODIES, ANTI-KLK5/KLK7 MULTI-SPECIFIC ANTIBODIES AND METHODS OF USE
IL305283A (en) 2021-03-15 2023-10-01 Genentech Inc Compositions and methods of treating lupus nephritis
EP4308242A1 (en) 2021-03-17 2024-01-24 Molecular Templates, Inc. Pd-l1 binding proteins comprising shiga toxin a subunit scaffolds and cd8+ t cell antigens
AU2022238571A1 (en) 2021-03-18 2023-09-14 Seagen Inc. Selective drug release from internalized conjugates of biologically active compounds
WO2022197877A1 (en) 2021-03-19 2022-09-22 Genentech, Inc. Methods and compositions for time delayed bio-orthogonal release of cytotoxic agents
JP2024510505A (en) 2021-03-19 2024-03-07 アイオバンス バイオセラピューティクス，インコーポレイテッド Methods for tumor-infiltrating lymphocyte (TIL) expansion and gene knockout in TILs associated with CD39/CD69 selection
CA3213080A1 (en) 2021-03-23 2022-09-29 Krit RITTHIPICHAI Cish gene editing of tumor infiltrating lymphocytes and uses of same in immunotherapy
EP4314253A2 (en) 2021-03-25 2024-02-07 Iovance Biotherapeutics, Inc. Methods and compositions for t-cell coculture potency assays and use with cell therapy products
US20240150466A1 (en) 2021-03-25 2024-05-09 Dynamicure Biotechnology Llc Anti-igfbp7 constructs and uses thereof
JP2024513837A (en) 2021-03-31 2024-03-27 バイオベラティブ・ユーエスエイ・インコーポレイテッド Reducing surgery-related hemolysis in patients with cold agglutinin disease
WO2022212876A1 (en) 2021-04-02 2022-10-06 The Regents Of The University Of California Antibodies against cleaved cdcp1 and uses thereof
CA3215049A1 (en) 2021-04-10 2022-10-13 Baiteng ZHAO Folr1 binding agents, conjugates thereof and methods of using the same
KR20230170738A (en) 2021-04-16 2023-12-19 노파르티스 아게 Antibody drug conjugate and method for producing the same
BR112023021665A2 (en) 2021-04-19 2023-12-19 Iovance Biotherapeutics Inc METHOD FOR TREATING A CANCER, AND, COMPOSITION
IL307501A (en) 2021-04-19 2023-12-01 Hoffmann La Roche Modified mammalian cells
WO2022226317A1 (en) 2021-04-23 2022-10-27 Profoundbio Us Co. Anti-cd70 antibodies, conjugates thereof and methods of using the same
WO2022235867A2 (en) 2021-05-06 2022-11-10 The Rockefeller University Neutralizing anti-sars- cov-2 antibodies and methods of use thereof
IL308198A (en) 2021-05-07 2024-01-01 Surface Oncology Llc Anti-il-27 antibodies and uses thereof
CA3216795A1 (en) 2021-05-07 2022-11-10 Alpine Immune Sciences, Inc. Methods of dosing and treatment with a taci-fc fusion immunomodulatory protein
EP4337266A1 (en) 2021-05-12 2024-03-20 Genentech, Inc. Methods of using anti-cd79b immunoconjugates to treat diffuse large b-cell lymphoma
AR125855A1 (en) 2021-05-14 2023-08-16 Genentech Inc TREM2 AGONISTS
JP2024519029A (en) 2021-05-17 2024-05-08 アイオバンス バイオセラピューティクス，インコーポレイテッド PD-1 gene-edited tumor-infiltrating lymphocytes and their use in immunotherapy
EP4341385A1 (en) 2021-05-21 2024-03-27 Genentech, Inc. Modified cells for the production of a recombinant product of interest
AR126054A1 (en) 2021-06-04 2023-09-06 Chugai Pharmaceutical Co Ltd ANTI-DDR2 ANTIBODIES AND USES OF THESE
JP2024522213A (en) 2021-06-14 2024-06-11 アルジェニクス ビーブイ Anti-IL-9 antibodies and methods of use thereof
US20240279334A1 (en) 2021-06-17 2024-08-22 Amberstone Biosciences, Inc. Anti-cd3 constructs and uses thereof
IL308959A (en) 2021-06-18 2024-01-01 Nammi Therapeutics Inc Fusion protein composition(s) comprising masked type i interferons (ifnα and ifnβ) for use in the treatment of cancer and methods thereof
TW202306989A (en) 2021-06-22 2023-02-16 瑞士商諾華公司 Bispecific antibodies for use in treatment of hidradenitis suppurativa
KR102690141B1 (en) 2021-06-25 2024-07-30 추가이 세이야쿠 가부시키가이샤 Anti-CTLA-4 antibody
JP7477127B2 (en) 2021-06-25 2024-05-01 中外製薬株式会社 Uses of Anti-CTLA-4 Antibodies
EP4363450A1 (en) 2021-07-01 2024-05-08 Compugen Ltd. Anti-tigit and anti-pvrig in monotherapy and combination treatments
TW202320857A (en) 2021-07-06 2023-06-01 美商普方生物製藥美國公司 Linkers, drug linkers and conjugates thereof and methods of using the same
CN118103397A (en) 2021-07-08 2024-05-28 舒泰神(加州)生物科技有限公司 Antibodies specifically recognizing TNFR2 and uses thereof
JP2024530402A (en) 2021-07-12 2024-08-21 ジェネンテック， インコーポレイテッド Structures for reducing antibody-lipase binding
EP4370550A1 (en) 2021-07-12 2024-05-22 INSERM (Institut National de la Santé et de la Recherche Médicale) Use of il-36 inhibitors for the treatment of netherton syndrome
TW202309097A (en) 2021-07-14 2023-03-01 美商建南德克公司 Anti-c-c motif chemokine receptor 8 (ccr8) antibodies and methods of use
CN115812082A (en) 2021-07-14 2023-03-17 舒泰神（北京）生物制药股份有限公司 Antibody specifically recognizing CD40 and application thereof
EP4373576A1 (en) 2021-07-22 2024-05-29 Genentech, Inc. Brain targeting compositions and methods of use thereof
EP4373270A2 (en) 2021-07-22 2024-05-29 Iovance Biotherapeutics, Inc. Method for cryopreservation of solid tumor fragments
KR20240036570A (en) 2021-07-22 2024-03-20 에프. 호프만-라 로슈 아게 Heterodimeric Fc domain antibodies
JP2024527977A (en) 2021-07-27 2024-07-26 ノヴァブ， インコーポレイテッド Engineered VLRB antibodies with immune effector functions
TW202327631A (en) 2021-07-28 2023-07-16 美商艾歐凡斯生物治療公司 Treatment of cancer patients with tumor infiltrating lymphocyte therapies in combination with kras inhibitors
EP4382538A1 (en) 2021-08-02 2024-06-12 Innovent Biologics (Suzhou) Co., Ltd. Anti-cd79b×cd3 bispecific antibody and use thereof
WO2023012147A1 (en) 2021-08-03 2023-02-09 F. Hoffmann-La Roche Ag Bispecific antibodies and methods of use
WO2023019092A1 (en) 2021-08-07 2023-02-16 Genentech, Inc. Methods of using anti-cd79b immunoconjugates to treat diffuse large b-cell lymphoma
CN117897409A (en) 2021-08-13 2024-04-16 基因泰克公司 Administration of anti-tryptase antibodies
CN117858905A (en) 2021-08-19 2024-04-09 豪夫迈·罗氏有限公司 Multivalent anti-variant FC region antibodies and methods of use
EP4392067A1 (en) 2021-08-27 2024-07-03 Genentech, Inc. Methods of treating tau pathologies
EP4396223A1 (en) 2021-08-30 2024-07-10 Genentech, Inc. Anti-polyubiquitin multispecific antibodies
EP4398915A1 (en) 2021-09-09 2024-07-17 Iovance Biotherapeutics, Inc. Processes for generating til products using pd-1 talen knockdown
WO2023049862A1 (en) 2021-09-24 2023-03-30 Iovance Biotherapeutics, Inc. Expansion processes and agents for tumor infiltrating lymphocytes
WO2023053282A1 (en) 2021-09-29 2023-04-06 中外製薬株式会社 Cytotoxicity-inducing therapeutic agent for use in treatment of cancer
WO2023052541A1 (en) 2021-09-30 2023-04-06 Imcheck Therapeutics Combination of an anti-btn3a activating antibody and an il-2 agonist for use in therapy
WO2023056403A1 (en) 2021-09-30 2023-04-06 Genentech, Inc. Methods for treatment of hematologic cancers using anti-tigit antibodies, anti-cd38 antibodies, and pd-1 axis binding antagonists
EP4408478A1 (en) 2021-09-30 2024-08-07 Seagen Inc. B7-h4 antibody-drug conjugates for the treatment of cancer
WO2023056069A1 (en) 2021-09-30 2023-04-06 Angiex, Inc. Degrader-antibody conjugates and methods of using same
CN116064598B (en) 2021-10-08 2024-03-12 苏州艾博生物科技有限公司 Nucleic acid vaccine for coronavirus
CN118176217A (en) 2021-10-14 2024-06-11 徕特康(苏州)生物制药有限公司 Novel antibody-cytokine fusion protein, and preparation method and application thereof
WO2023064958A1 (en) 2021-10-15 2023-04-20 Compugen Ltd. Combination therapy with anti-pvrig antibodies formulations, anti-tigit antibodies, and anti-pd-1 antibodies
WO2023076876A1 (en) 2021-10-26 2023-05-04 Mozart Therapeutics, Inc. Modulation of immune responses to viral vectors
AR127482A1 (en) 2021-10-27 2024-01-31 Iovance Biotherapeutics Inc SYSTEMS AND METHODS TO COORDINATE THE MANUFACTURE OF CELLS FOR PATIENT-SPECIFIC IMMUNOTHERAPY
TW202342095A (en) 2021-11-05 2023-11-01 英商阿斯特捷利康英國股份有限公司 Composition for treatment and prevention of covid-19
EP4426748A1 (en) 2021-11-05 2024-09-11 American Diagnostics & Therapy, LLC (ADXRX) Monoclonal antibodies against carcinoembryonic antigens, and their uses
WO2023086807A1 (en) 2021-11-10 2023-05-19 Genentech, Inc. Anti-interleukin-33 antibodies and uses thereof
AU2022388729A1 (en) 2021-11-10 2024-05-16 Iovance Biotherapeutics, Inc. Methods of expansion treatment utilizing cd8 tumor infiltrating lymphocytes
TW202337494A (en) 2021-11-16 2023-10-01 美商建南德克公司 Methods and compositions for treating systemic lupus erythematosus (sle) with mosunetuzumab
WO2023088959A1 (en) 2021-11-16 2023-05-25 Ac Immune Sa Novel molecules for therapy and diagnosis
CA3235096A1 (en) 2021-11-17 2023-05-25 Disc Medicine, Inc. Methods for treating anemia of kidney disease
KR20240112934A (en) 2021-12-01 2024-07-19 인쎄름 (엥스띠뛰 나씨오날 드 라 쌍떼 에 드 라 흐쉐르슈 메디깔) Neutralizing anti-CD95L monoclonal antibody
CN116829179A (en) 2021-12-06 2023-09-29 北京三诺佳邑生物技术有限责任公司 Bispecific antibodies and compositions that specifically bind Klebsiella pneumoniae O2 antigen and O1 antigen
AU2022414067A1 (en) 2021-12-15 2024-05-23 Genentech, Inc. Stabilized il-18 polypeptides and uses thereof
WO2023109901A1 (en) 2021-12-17 2023-06-22 Shanghai Henlius Biotech, Inc. Anti-ox40 antibodies and methods of use
CN118488965A (en) 2021-12-17 2024-08-13 上海复宏汉霖生物技术股份有限公司 Anti-OX 40 antibodies, multispecific antibodies, and methods of use thereof
AR128222A1 (en) 2022-01-07 2024-04-10 Johnson & Johnson Entpr Innovation Inc IL-1b BINDING PROTEIN MATERIALS AND METHODS
AU2023208882A1 (en) 2022-01-18 2024-05-09 argenx BV Galectin-10 antibodies
WO2023141445A1 (en) 2022-01-19 2023-07-27 Genentech, Inc. Anti-notch2 antibodies and conjugates and methods of use
WO2023147399A1 (en) 2022-01-27 2023-08-03 The Rockefeller University Broadly neutralizing anti-sars-cov-2 antibodies targeting the n-terminal domain of the spike protein and methods of use thereof
WO2023147488A1 (en) 2022-01-28 2023-08-03 Iovance Biotherapeutics, Inc. Cytokine associated tumor infiltrating lymphocytes compositions and methods
WO2023147486A1 (en) 2022-01-28 2023-08-03 Iovance Biotherapeutics, Inc. Tumor infiltrating lymphocytes engineered to express payloads
AU2023217319A1 (en) 2022-02-08 2024-08-22 Assistance Publique-Hôpitaux De Paris (Aphp) Neutralization of acyl-coa binding protein confers autophagy-dependent organ protection
EP4227321A1 (en) 2022-02-10 2023-08-16 AVA Lifescience GmbH Antibodies targeting the b-cell receptor of chronic lymphocytic leukemia (cll) for use in treatment of cll
WO2023152204A1 (en) 2022-02-10 2023-08-17 Ava Lifescience Gmbh Antibodies targeting the b-cell receptor of chronic lymphocytic leukemia (cll) for use in treatment of cll
WO2023156437A1 (en) 2022-02-16 2023-08-24 INSERM (Institut National de la Santé et de la Recherche Médicale) Anti-robo4 human monoclonal antibodies and uses thereof for the treatment of cancer
TW202342510A (en) 2022-02-18 2023-11-01 英商Ｒｑ生物科技有限公司 Antibodies
WO2023156587A1 (en) 2022-02-18 2023-08-24 INSERM (Institut National de la Santé et de la Recherche Médicale) Use of tcr-deficient car-tregs in combination with anti-tcr complex monoclonal antibodies for inducing durable tolerance
WO2023161412A1 (en) 2022-02-25 2023-08-31 INSERM (Institut National de la Santé et de la Recherche Médicale) Antibodies for preventing the cleavage of cd95l by metalloproteases
WO2023172883A1 (en) 2022-03-07 2023-09-14 Alpine Immune Sciences, Inc. Immunomodulatory proteins of variant cd80 polypeptides, cell therapies thereof and related methods and uses
WO2023170207A1 (en) 2022-03-09 2023-09-14 Alderaan Biotechnology Anti-cd160 transmembrane isoform antibodies
AU2023236289A1 (en) 2022-03-15 2024-08-15 Compugen Ltd. Il-18bp antagonist antibodies and their use in monotherapy and combination therapy in the treatment of cancer
EP4245374A3 (en) 2022-03-18 2024-07-10 Compugen Ltd. Pvrl2 and/or pvrig as biomarkers for treatment
WO2023178357A1 (en) 2022-03-18 2023-09-21 Evolveimmune Therapeutics, Inc. Bispecific antibody fusion molecules and methods of use thereof
AU2023240941A1 (en) 2022-03-25 2024-09-19 Shanghai Henlius Biologics Co., Ltd. Anti-msln antibodies and methods of use
US20230302423A1 (en) 2022-03-28 2023-09-28 Massachusetts Institute Of Technology Rna scaffolded wireframe origami and methods thereof
AU2023247309A1 (en) 2022-03-30 2024-09-12 Novartis Ag Methods of treating disorders using anti-natriuretic peptide receptor 1 (npr1) antibodies
WO2023192802A1 (en) 2022-04-01 2023-10-05 The Board Of Trustees Of The University Of Illinois Notch1 and notch4 decoys and methods of use
WO2023191816A1 (en) 2022-04-01 2023-10-05 Genentech, Inc. Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies
US12071621B2 (en) 2022-04-05 2024-08-27 Avidity Biosciences, Inc. Anti-transferrin receptor antibody-PMO conjugates for inducing DMD exon 44 skipping
WO2023196877A1 (en) 2022-04-06 2023-10-12 Iovance Biotherapeutics, Inc. Treatment of nsclc patients with tumor infiltrating lymphocyte therapies
TW202408562A (en) 2022-04-13 2024-03-01 美商建南德克公司 Pharmaceutical compositions of therapeutic proteins and methods of use
WO2023201369A1 (en) 2022-04-15 2023-10-19 Iovance Biotherapeutics, Inc. Til expansion processes using specific cytokine combinations and/or akti treatment
CN115322255A (en) * 2022-04-25 2022-11-11 上海药明生物医药有限公司 Optimized Fc variant and application thereof
WO2023209177A1 (en) 2022-04-29 2023-11-02 Astrazeneca Uk Limited Sars-cov-2 antibodies and methods of using the same
TW202406934A (en) 2022-05-03 2024-02-16 美商建南德克公司 Anti-ly6e antibodies, immunoconjugates, and uses thereof
WO2023220608A1 (en) 2022-05-10 2023-11-16 Iovance Biotherapeutics, Inc. Treatment of cancer patients with tumor infiltrating lymphocyte therapies in combination with an il-15r agonist
TW202400638A (en) 2022-05-10 2024-01-01 法商感應檢查療法公司 Anti-btn3a antibodies for use in methods of treating gastro-intestinal inflammatory disorders
AR129268A1 (en) 2022-05-11 2024-08-07 Hoffmann La Roche ANTIBODY THAT BINDS TO VEGF-A AND IL6 AND METHODS OF USE
WO2023219613A1 (en) 2022-05-11 2023-11-16 Genentech, Inc. Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies
WO2023222557A1 (en) 2022-05-17 2023-11-23 Bayer Aktiengesellschaft Radiopharmaceutical complexes targeting prostate-specific membrane antigen and its combinations
EP4279092A1 (en) 2022-05-17 2023-11-22 Bayer AG Radiopharmaceutical complexes targeting prostate-specific membrane antigen
WO2023228082A1 (en) 2022-05-26 2023-11-30 Pfizer Inc. Anti-tnfr2 antibodies and methods of use thereof
TW202405002A (en) 2022-05-31 2024-02-01 美商輝瑞大藥廠 Anti-bmp9 antibodies and methods of use thereof
WO2023235699A1 (en) 2022-05-31 2023-12-07 Jounce Therapeutics, Inc. Antibodies to lilrb4 and uses thereof
WO2023240058A2 (en) 2022-06-07 2023-12-14 Genentech, Inc. Prognostic and therapeutic methods for cancer
WO2023239803A1 (en) 2022-06-08 2023-12-14 Angiex, Inc. Anti-tm4sf1 antibody-drug conjugates comprising cleavable linkers and methods of using same
TW202417039A (en) 2022-06-15 2024-05-01 美商百歐維拉提夫美國公司 Anti-complement c1s antibody formulation
WO2023242372A1 (en) 2022-06-15 2023-12-21 argenx BV Fcrn/hsa binding molecules and methods of use
US20240025978A1 (en) 2022-06-24 2024-01-25 Bioverativ Usa Inc. Methods for treating complement-mediated diseases
WO2024011114A1 (en) 2022-07-06 2024-01-11 Iovance Biotherapeutics, Inc. Devices and processes for automated production of tumor infiltrating lymphocytes
WO2024015897A1 (en) 2022-07-13 2024-01-18 Genentech, Inc. Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies
TW202413433A (en) 2022-07-19 2024-04-01 美商建南德克公司 Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies
TW202417503A (en) 2022-07-19 2024-05-01 美商舒泰神（加州）生物科技有限公司 Antibodies specifically recognizing b- and t-lymphocyte attenuator (btla) and uses thereof
EP4309733A1 (en) 2022-07-22 2024-01-24 Institut National de la Santé et de la Recherche Médicale (INSERM) Neutralization of acyl-coa binding protein for the treatment of cardiac dysfunction
WO2024020564A1 (en) 2022-07-22 2024-01-25 Genentech, Inc. Anti-steap1 antigen-binding molecules and uses thereof
WO2024020579A1 (en) 2022-07-22 2024-01-25 Bristol-Myers Squibb Company Antibodies binding to human pad4 and uses thereof
WO2024026496A1 (en) 2022-07-28 2024-02-01 Compugen Ltd. Combination therapy with anti-pvrig antibodies formulations and anti-pd-1 antibodies
WO2024030758A1 (en) 2022-08-01 2024-02-08 Iovance Biotherapeutics, Inc. Chimeric costimulatory receptors, chemokine receptors, and the use of same in cellular immunotherapies
US20240052047A1 (en) 2022-08-03 2024-02-15 Pfizer Inc. Anti- il27r antibodies and methods of use thereof
WO2024030956A2 (en) 2022-08-03 2024-02-08 Mozart Therapeutics, Inc. Cd39-specific binding agents and methods of using the same
WO2024028731A1 (en) 2022-08-05 2024-02-08 Janssen Biotech, Inc. Transferrin receptor binding proteins for treating brain tumors
WO2024028732A1 (en) 2022-08-05 2024-02-08 Janssen Biotech, Inc. Cd98 binding constructs for treating brain tumors
WO2024037633A2 (en) 2022-08-19 2024-02-22 Evive Biotechnology (Shanghai) Ltd Formulations comprising g-csf and uses thereof
WO2024044675A1 (en) 2022-08-25 2024-02-29 Beigene, Ltd. Methods of cancer treatment using anti-pd1 antibodies in combination with anti-tim3 antibodies
WO2024050524A1 (en) 2022-09-01 2024-03-07 University Of Georgia Research Foundation, Inc. Compositions and methods for directing apolipoprotein l1 to induce mammalian cell death
WO2024049949A1 (en) 2022-09-01 2024-03-07 Genentech, Inc. Therapeutic and diagnostic methods for bladder cancer
WO2024054929A1 (en) 2022-09-07 2024-03-14 Dynamicure Biotechnology Llc Anti-vista constructs and uses thereof
WO2024052503A1 (en) 2022-09-08 2024-03-14 Institut National de la Santé et de la Recherche Médicale Antibodies having specificity to ltbp2 and uses thereof
WO2024064752A2 (en) 2022-09-20 2024-03-28 Yale University Compositions of wet adhesives derived from vibrio cholerae biofilm adhesins and methods thereof
WO2024062074A1 (en) 2022-09-21 2024-03-28 Sanofi Biotechnology Humanized anti-il-1r3 antibody and methods of use
WO2024074498A1 (en) 2022-10-04 2024-04-11 Imcheck Therapeutics Combination of a btn3a activating antibody, a bcl2 inhibitor and hypomethylating agent for use in treating cancer
WO2024077018A2 (en) 2022-10-04 2024-04-11 Alpine Immune Sciences, Inc. Methods and uses of taci-fc fusion immunomodulatory protein
WO2024077239A1 (en) 2022-10-07 2024-04-11 Genentech, Inc. Methods of treating cancer with anti-c-c motif chemokine receptor 8 (ccr8) antibodies
WO2024083790A1 (en) 2022-10-17 2024-04-25 Institut Gustave Roussy Cd207 dendritic cells as a biomarker for tumoural progression and therapeutic target
WO2024091991A1 (en) 2022-10-25 2024-05-02 Genentech, Inc. Therapeutic and diagnostic methods for multiple myeloma
WO2024089609A1 (en) 2022-10-25 2024-05-02 Ablynx N.V. Glycoengineered fc variant polypeptides with enhanced effector function
WO2024094741A1 (en) 2022-11-03 2024-05-10 F. Hoffmann-La Roche Ag Combination therapy with anti-cd19/anti-cd28 bispecific antibody
WO2024097741A1 (en) 2022-11-04 2024-05-10 Gilead Sciences, Inc. Anticancer therapies using anti-ccr8 antibody, chemo and immunotherapy combinations
WO2024098024A1 (en) 2022-11-04 2024-05-10 Iovance Biotherapeutics, Inc. Expansion of tumor infiltrating lymphocytes from liquid tumors and therapeutic uses thereof
WO2024098027A1 (en) 2022-11-04 2024-05-10 Iovance Biotherapeutics, Inc. Methods for tumor infiltrating lymphocyte (til) expansion related to cd39/cd103 selection
WO2024102734A1 (en) 2022-11-08 2024-05-16 Genentech, Inc. Compositions and methods of treating childhood onset idiopathic nephrotic syndrome
WO2024107752A2 (en) 2022-11-15 2024-05-23 Onestone Therapeutics Llc Compositions and methods for immunomodulatory bifunctional fusion molecules
US20240226313A1 (en) 2022-11-17 2024-07-11 Sanofi Ceacam5 antibody-drug conjugates and methods of use thereof
WO2024112571A2 (en) 2022-11-21 2024-05-30 Iovance Biotherapeutics, Inc. Two-dimensional processes for the expansion of tumor infiltrating lymphocytes and therapies therefrom
WO2024112711A2 (en) 2022-11-21 2024-05-30 Iovance Biotherapeutics, Inc. Methods for assessing proliferation potency of gene-edited t cells
WO2024110905A1 (en) 2022-11-24 2024-05-30 Beigene, Ltd. Anti-cea antibody drug conjugates and methods of use
EP4382120A1 (en) 2022-12-05 2024-06-12 Institut Regional du Cancer de Montpellier Anti-slc1a4 monoclonal antibodies and uses thereof
WO2024145398A1 (en) 2022-12-27 2024-07-04 Yale University Antibody drug conjugates
WO2024150074A2 (en) 2023-01-13 2024-07-18 Takeda Pharmaceutical Company Limited Coronavirus antibodies and therapeutic uses thereof
WO2024151885A1 (en) 2023-01-13 2024-07-18 Iovance Biotherapeutics, Inc. Use of til as maintenance therapy for nsclc patients who achieved pr/cr after prior therapy
WO2024155807A1 (en) 2023-01-18 2024-07-25 Genentech, Inc. Multispecific antibodies and uses thereof
WO2024153168A2 (en) 2023-01-19 2024-07-25 Beigene, Ltd. Anti-cmet antibodies and methods of use
WO2024158824A1 (en) 2023-01-23 2024-08-02 Yale University Antibody oligonucleotide conjugates
WO2024170543A1 (en) 2023-02-14 2024-08-22 Institut National de la Santé et de la Recherche Médicale Anti-cd44 antibodies and uses thereof
WO2024173607A2 (en) 2023-02-14 2024-08-22 Evolveimmune Therapeutics, Inc. Combination of bispecific antibodies and chimeric antigen receptor t cells for treatment
WO2024170756A1 (en) 2023-02-17 2024-08-22 Ablynx N.V. Polypeptides binding to the neonatal fc receptor
WO2024175699A1 (en) 2023-02-23 2024-08-29 Imcheck Therapeutics Combination of btn3a activating antibody and immune checkpoint inhibitors
WO2024182443A2 (en) 2023-02-27 2024-09-06 Compugen Ltd. Triple combination therapy with anti-pvrig antibodies, anti-tigit antibodies, and pembrolizumab
WO2024182714A1 (en) 2023-03-02 2024-09-06 Alloy Therapeutics, Inc. Anti-cd22 antibodies and uses thereof
WO2024183636A1 (en) 2023-03-03 2024-09-12 Beigene Switzerland Gmbh Cd16a antibodies and methods of use
WO2024183635A1 (en) 2023-03-03 2024-09-12 Beigene, Ltd. Muc1 and cd16a antibodies and methods of use
WO2024186635A2 (en) 2023-03-03 2024-09-12 Celldex Therapeutics, Inc. Anti-stem cell factor (scf) and anti-thymic stromal lymphopoietin (tslp) antibodies and bispecific constructs
WO2024183637A1 (en) 2023-03-03 2024-09-12 Beigene Switzerland Gmbh Muc1 antibodies and methods of use
WO2024184812A1 (en) 2023-03-06 2024-09-12 Beigene Switzerland Gmbh Anti-cldn6 antibodies and methods of use
WO2024184810A1 (en) 2023-03-06 2024-09-12 Beigene Switzerland Gmbh Anti-cldn6 and anti-cd3 multispecific antibodies and methods of use
WO2024184811A1 (en) 2023-03-06 2024-09-12 Beigene Switzerland Gmbh Anti-cd3 multispecific antibodies and methods of use

Citations (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US98193A (en) * 1869-12-21 Improvement in corn-planters
US158389A (en) * 1875-01-05 Improvement in gas-regulators
US5419904A (en) * 1990-11-05 1995-05-30 The Regents Of The University Of California Human B-lymphoblastoid cell line secreting anti-ganglioside antibody
US5576184A (en) * 1988-09-06 1996-11-19 Xoma Corporation Production of chimeric mouse-human antibodies with specificity to human tumor antigens
US5731168A (en) * 1995-03-01 1998-03-24 Genentech, Inc. Method for making heteromultimeric polypeptides
US5985599A (en) * 1986-05-29 1999-11-16 The Austin Research Institute FC receptor for immunoglobulin
US6165745A (en) * 1992-04-24 2000-12-26 Board Of Regents, The University Of Texas System Recombinant production of immunoglobulin-like domains in prokaryotic cells
US6277375B1 (en) * 1997-03-03 2001-08-21 Board Of Regents, The University Of Texas System Immunoglobulin-like domains with increased half-lives
US20030036459A1 (en) * 2001-08-16 2003-02-20 Jeon Ig-Kyun Brake system of vehicle
US6528624B1 (en) * 1998-04-02 2003-03-04 Genentech, Inc. Polypeptide variants
US6538124B1 (en) * 1998-04-02 2003-03-25 Genentech, Inc. Polypeptide variants
US20040002587A1 (en) * 2002-02-20 2004-01-01 Watkins Jeffry D. Fc region variants
US6676927B1 (en) * 1999-01-20 2004-01-13 The Rockefeller University Animal model and methods for its use in the selection of cytotoxic antibodies
US6737056B1 (en) * 1999-01-15 2004-05-18 Genentech, Inc. Polypeptide variants with altered effector function

Family Cites Families (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4752601A (en) 1983-08-12 1988-06-21 Immunetech Pharmaceuticals Method of blocking immune complex binding to immunoglobulin FC receptors
EP0307434B2 (en) 1987-03-18 1998-07-29 Scotgen Biopharmaceuticals, Inc. Altered antibodies
GB8916400D0 (en) 1989-07-18 1989-09-06 Dynal As Modified igg3
US5364930A (en) 1990-10-16 1994-11-15 Northwestern University Synthetic C1q peptide fragments
US5736137A (en) 1992-11-13 1998-04-07 Idec Pharmaceuticals Corporation Therapeutic application of chimeric and radiolabeled antibodies to human B lymphocyte restricted differentiation antigen for treatment of B cell lymphoma
CA2163345A1 (en) 1993-06-16 1994-12-22 Susan Adrienne Morgan Antibodies
US6750334B1 (en) 1996-02-02 2004-06-15 Repligen Corporation CTLA4-immunoglobulin fusion proteins having modified effector functions and uses therefor
US5834597A (en) 1996-05-20 1998-11-10 Protein Design Labs, Inc. Mutated nonactivating IgG2 domains and anti CD3 antibodies incorporating the same
WO1998023289A1 (en) 1996-11-27 1998-06-04 The General Hospital Corporation MODULATION OF IgG BINDING TO FcRn
DE19721700C1 (en) 1997-05-23 1998-11-19 Deutsches Krebsforsch Mutant OKT3 antibody
CN1204147C (en) 1998-02-25 2005-06-01 利思进药品公司 Enhancing circulating half-life of antibody-based fusion proteins
ES2292236T3 (en) 1998-04-02 2008-03-01 Genentech, Inc. VARIATIONS OF ANTIBODIES AND THEIR FRAGMENTS.
GB9809951D0 (en) 1998-05-08 1998-07-08 Univ Cambridge Tech Binding molecules
WO2000009560A2 (en) 1998-08-17 2000-02-24 Abgenix, Inc. Generation of modified molecules with increased serum half-lives

Patent Citations (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US158389A (en) * 1875-01-05 Improvement in gas-regulators
US98193A (en) * 1869-12-21 Improvement in corn-planters
US5985599A (en) * 1986-05-29 1999-11-16 The Austin Research Institute FC receptor for immunoglobulin
US5576184A (en) * 1988-09-06 1996-11-19 Xoma Corporation Production of chimeric mouse-human antibodies with specificity to human tumor antigens
US5419904A (en) * 1990-11-05 1995-05-30 The Regents Of The University Of California Human B-lymphoblastoid cell line secreting anti-ganglioside antibody
US6165745A (en) * 1992-04-24 2000-12-26 Board Of Regents, The University Of Texas System Recombinant production of immunoglobulin-like domains in prokaryotic cells
US5731168A (en) * 1995-03-01 1998-03-24 Genentech, Inc. Method for making heteromultimeric polypeptides
US6277375B1 (en) * 1997-03-03 2001-08-21 Board Of Regents, The University Of Texas System Immunoglobulin-like domains with increased half-lives
US6821505B2 (en) * 1997-03-03 2004-11-23 Board Of Regents, The University Of Texas System Immunoglobin-like domains with increased half lives
US6528624B1 (en) * 1998-04-02 2003-03-04 Genentech, Inc. Polypeptide variants
US6538124B1 (en) * 1998-04-02 2003-03-25 Genentech, Inc. Polypeptide variants
US20040191244A1 (en) * 1999-01-15 2004-09-30 Genentech, Inc. Polypeptide variants with altered effector function
US20040228856A1 (en) * 1999-01-15 2004-11-18 Genentech, Inc. Polypeptide variants with altered effector function
US6737056B1 (en) * 1999-01-15 2004-05-18 Genentech, Inc. Polypeptide variants with altered effector function
US6676927B1 (en) * 1999-01-20 2004-01-13 The Rockefeller University Animal model and methods for its use in the selection of cytotoxic antibodies
US20030036459A1 (en) * 2001-08-16 2003-02-20 Jeon Ig-Kyun Brake system of vehicle
US20040002587A1 (en) * 2002-02-20 2004-01-01 Watkins Jeffry D. Fc region variants

Cited By (161)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030158389A1 (en) * 1998-04-02 2003-08-21 Genentech, Inc. Polypeptide variants
US7364731B2 (en) 1998-04-02 2008-04-29 Genentech, Inc. Polypeptide variants
US7297775B2 (en) 1998-04-02 2007-11-20 Genentech, Inc. Polypeptide variants
US7741072B2 (en) 1998-04-02 2010-06-22 Genentech, Inc. Polypeptide variants
US20060194954A1 (en) * 1998-04-02 2006-08-31 Genentech, Inc. Polypeptide variants
US20080274105A1 (en) * 1999-01-15 2008-11-06 Genentech, Inc. Polypeptide variants with altered effector function
US7335742B2 (en) 1999-01-15 2008-02-26 Genentech, Inc. Polypeptide variants with altered effector function
US8674083B2 (en) 1999-01-15 2014-03-18 Genentech, Inc. Polypeptide variants with altered effector function
US7332581B2 (en) 1999-01-15 2008-02-19 Genentech, Inc. Polypeptide variants with altered effector function
US8163882B2 (en) 1999-01-15 2012-04-24 Genentech, Inc. Polypeptide variants with altered effector function
US7371826B2 (en) 1999-01-15 2008-05-13 Genentech, Inc. Polypeptide variants with altered effector function
US20040191244A1 (en) * 1999-01-15 2004-09-30 Genentech, Inc. Polypeptide variants with altered effector function
US20050118174A1 (en) * 1999-01-15 2005-06-02 Genentech, Inc. Polypeptide variants with altered effector function
US7416727B2 (en) 1999-01-15 2008-08-26 Genentech, Inc. Polypeptide variants with altered effector function
US20060194290A1 (en) * 1999-01-15 2006-08-31 Genentech, Inc. Polypeptide variants with altered effector function
US20060194957A1 (en) * 1999-01-15 2006-08-31 Genentech, Inc. Polypeptide variants with altered effector function
US20060194291A1 (en) * 1999-01-15 2006-08-31 Genentech, Inc. Polypeptide variants with altered effector function
US20080274506A1 (en) * 1999-01-15 2008-11-06 Genentech, Inc. Polypeptide variants with altered effector function
US7122637B2 (en) 1999-01-15 2006-10-17 Genentech, Inc. Polypeptide variants with altered effector function
US7790858B2 (en) 1999-01-15 2010-09-07 Genentech, Inc. Polypeptide variants with altered effector function
US7785791B2 (en) 1999-01-15 2010-08-31 Genentech, Inc. Polypeptide variants with altered effector function
US20040228856A1 (en) * 1999-01-15 2004-11-18 Genentech, Inc. Polypeptide variants with altered effector function
US20080274108A1 (en) * 1999-01-15 2008-11-06 Genentech, Inc. Polypeptide variants with altered effector function
US7183387B1 (en) 1999-01-15 2007-02-27 Genentech, Inc. Polypeptide variants with altered effector function
US8545843B2 (en) 1999-05-07 2013-10-01 Genentech, Inc. Treatment of vasculitis
US9993550B2 (en) 1999-05-07 2018-06-12 Genentech, Inc. Treatment of pemphigus
US7820161B1 (en) 1999-05-07 2010-10-26 Biogen Idec, Inc. Treatment of autoimmune diseases
US8853366B2 (en) 2001-01-17 2014-10-07 Emergent Product Development Seattle, Llc Binding domain-immunoglobulin fusion proteins
US20080260731A1 (en) * 2002-03-01 2008-10-23 Bernett Matthew J Optimized antibodies that target cd19
US20070003546A1 (en) * 2002-03-01 2007-01-04 Xencor, Inc. Optimized Fc variants and methods for their generation
US8734791B2 (en) 2002-03-01 2014-05-27 Xencor, Inc. Optimized fc variants and methods for their generation
US20090068175A1 (en) * 2002-03-01 2009-03-12 Xencor, Inc. Optimized FC Variants and Methods for Their Generation
US7662925B2 (en) 2002-03-01 2010-02-16 Xencor, Inc. Optimized Fc variants and methods for their generation
US20080254027A1 (en) * 2002-03-01 2008-10-16 Bernett Matthew J Optimized CD5 antibodies and methods of using the same
US20090142340A1 (en) * 2002-03-01 2009-06-04 Xencor, Inc. Optimized Fc Variants and Methods for Their Generation
US8093357B2 (en) 2002-03-01 2012-01-10 Xencor, Inc. Optimized Fc variants and methods for their generation
US20080181890A1 (en) * 2002-03-01 2008-07-31 Xencor, Inc. Optimized Fc Variants and Methods for Their Generation
US8124731B2 (en) 2002-03-01 2012-02-28 Xencor, Inc. Optimized Fc variants and methods for their generation
US7317091B2 (en) 2002-03-01 2008-01-08 Xencor, Inc. Optimized Fc variants
US20070224189A1 (en) * 2002-03-01 2007-09-27 Xencor, Inc. CD20 OPTIMIZED Fc VARIANTS AND METHODS FOR THEIR GENERATION
US20070219133A1 (en) * 2002-03-01 2007-09-20 Xencor, Inc. CD52 OPTIMIZED Fc VARIANTS AND METHODS FOR THEIR GENERATION
US8039592B2 (en) 2002-09-27 2011-10-18 Xencor, Inc. Optimized Fc variants and methods for their generation
US20090092599A1 (en) * 2002-09-27 2009-04-09 Xencor, Inc. Optimized Fc variants and methods for their generation
US8753629B2 (en) 2002-09-27 2014-06-17 Xencor, Inc. Optimized Fc variants
US8735547B2 (en) 2002-09-27 2014-05-27 Xencor, Inc. Optimized Fc Variants
US8802823B2 (en) 2002-09-27 2014-08-12 Xencor, Inc. Optimized Fc variants
US8809503B2 (en) 2002-09-27 2014-08-19 Xencor, Inc. Optimized Fc variants and methods for their generation
US8858937B2 (en) 2002-09-27 2014-10-14 Xencor, Inc. Optimized Fc variants and methods for their generation
US8383109B2 (en) 2002-09-27 2013-02-26 Xencor, Inc. Optimized Fc variants and methods for their generation
US9193798B2 (en) 2002-09-27 2015-11-24 Xencor, Inc. Optimized Fc variants and methods for their generation
US9353187B2 (en) 2002-09-27 2016-05-31 Xencor, Inc. Optimized FC variants and methods for their generation
US8188231B2 (en) 2002-09-27 2012-05-29 Xencor, Inc. Optimized FC variants
US8093359B2 (en) 2002-09-27 2012-01-10 Xencor, Inc. Optimized Fc variants and methods for their generation
US20040132101A1 (en) * 2002-09-27 2004-07-08 Xencor Optimized Fc variants and methods for their generation
US20060235208A1 (en) * 2002-09-27 2006-10-19 Xencor, Inc. Fc variants with optimized properties
US10184000B2 (en) 2002-09-27 2019-01-22 Xencor, Inc. Optimized Fc variants and methods for their generation
US20070148171A1 (en) * 2002-09-27 2007-06-28 Xencor, Inc. Optimized anti-CD30 antibodies
US10183999B2 (en) 2002-09-27 2019-01-22 Xencor, Inc. Optimized Fc variants and methods for their generation
US8753628B2 (en) 2002-09-27 2014-06-17 Xencor, Inc. Optimized Fc variants
US20090081208A1 (en) * 2002-09-27 2009-03-26 Xencor, Inc. Optimized Fc variants and methods for their generation
US20070238665A1 (en) * 2003-03-03 2007-10-11 Xencor, Inc. Fc Variants Having Decreased Affinity for FcyRIIc
US9663582B2 (en) 2003-03-03 2017-05-30 Xencor, Inc. Optimized Fc variants
US8735545B2 (en) 2003-03-03 2014-05-27 Xencor, Inc. Fc variants having increased affinity for fcyrllc
US20070275460A1 (en) * 2003-03-03 2007-11-29 Xencor.Inc. Fc Variants With Optimized Fc Receptor Binding Properties
US10584176B2 (en) 2003-03-03 2020-03-10 Xencor, Inc. Fc variants with increased affinity for FcγRIIc
US20070248602A1 (en) * 2003-03-03 2007-10-25 Xencor, Inc. Fc Variants Having Increased Affinity for FcyRllc
US20070237766A1 (en) * 2003-03-03 2007-10-11 Xencor, Inc. Fc Variants Having Increased Affinity for FcyRllla
US10113001B2 (en) 2003-03-03 2018-10-30 Xencor, Inc. Fc variants with increased affinity for FcyRIIc
US8388955B2 (en) 2003-03-03 2013-03-05 Xencor, Inc. Fc variants
US20070248603A1 (en) * 2003-03-03 2007-10-25 Xencor, Inc. Fc Variants with Increased Affinity for FcyRlla
US20070237767A1 (en) * 2003-03-03 2007-10-11 Xencor, Inc. Fc Variants Having Decreased Affinity for FcyRllla
US9657106B2 (en) 2003-03-03 2017-05-23 Xencor, Inc. Optimized Fc variants
US20080057056A1 (en) * 2003-03-03 2008-03-06 Xencor, Inc. Fc Variants with Increased Affinity for FcyRIIC
US20070237765A1 (en) * 2003-03-03 2007-10-11 Xencor, Inc. Fc Variants Having Increased Affinity for FcyRl
US20070243188A1 (en) * 2003-03-03 2007-10-18 Xencor, Inc. Fc Variants Having Decreased Affinity for FcyRlla
US20110021755A1 (en) * 2003-03-03 2011-01-27 Xencor, Inc. Optimized Fc Variants
US8084582B2 (en) 2003-03-03 2011-12-27 Xencor, Inc. Optimized anti-CD20 monoclonal antibodies having Fc variants
US9051373B2 (en) 2003-05-02 2015-06-09 Xencor, Inc. Optimized Fc variants
US20050069521A1 (en) * 2003-08-28 2005-03-31 Emd Lexigen Research Center Corp. Enhancing the circulating half-life of interleukin-2 proteins
US9714282B2 (en) 2003-09-26 2017-07-25 Xencor, Inc. Optimized Fc variants and methods for their generation
US20100203046A1 (en) * 2003-11-12 2010-08-12 Biogen Idec Ma Inc. Fc gamma receptor-binding polypeptide variants and methods related thereto
US20060275283A1 (en) * 2003-11-12 2006-12-07 Biogen Idec Ma Inc. Fcgamma receptor-binding polypeptide variants and methods related thereto
US20100093979A1 (en) * 2003-12-22 2010-04-15 Gregory Alan Lazar Fc Polypeptides With Novel Fc Ligand Binding Sites
US20050249723A1 (en) * 2003-12-22 2005-11-10 Xencor, Inc. Fc polypeptides with novel Fc ligand binding sites
US8338575B2 (en) 2003-12-30 2012-12-25 Merck Patent Gmbh IL-7 fusion proteins
US7323549B2 (en) 2003-12-30 2008-01-29 Emd Lexigen Research Center Corp. IL-7 fusion proteins
US20050164352A1 (en) * 2003-12-30 2005-07-28 Emd Lexigen Research Center Corp. IL-7 fusion proteins
US20090010875A1 (en) * 2003-12-30 2009-01-08 Scott Lauder IL-7 Fusion Proteins
US7960514B2 (en) 2003-12-30 2011-06-14 Merck Patent Gmbh IL-7 fusion proteins
US7432357B2 (en) 2004-01-22 2008-10-07 Merck Patent Gmbh Anti-cancer antibodies with reduced complement fixation
JP2007537719A (en) * 2004-01-22 2007-12-27 メルク　パテント　ゲゼルシャフト　ミット　ベシュレンクテル　ハフトング Anticancer antibodies with reduced complement binding
US9617349B2 (en) 2004-01-22 2017-04-11 Merck Patent Gmbh Anti-cancer antibodies with reduced complement fixation
US10017579B2 (en) 2004-01-22 2018-07-10 Meck Patent Gmbh Anti-cancer antibodies with reduced complement fixation
US20090148441A1 (en) * 2004-01-22 2009-06-11 Merck Patent Gmbh Anti-Cancer Antibodies With Reduced Complement Fixation
WO2005070967A2 (en) * 2004-01-22 2005-08-04 Merck Patent Gmbh Anti-cancer antibodies with reduced complement fixation
WO2005070967A3 (en) * 2004-01-22 2005-12-01 Merck Patent Gmbh Anti-cancer antibodies with reduced complement fixation
US10633452B2 (en) 2004-01-22 2020-04-28 Merck Patent Gmbh Anti-cancer antibodies with reduced complement fixation
US20050202021A1 (en) * 2004-01-22 2005-09-15 Emd Lexigen Research Center Corp. Anti-cancer antibodies with reduced complement fixation
US8835606B2 (en) 2004-01-22 2014-09-16 Merck Patent Gmbh Anti-cancer antibodies with reduced complement fixation
US20050244403A1 (en) * 2004-03-24 2005-11-03 Xencor, Inc. Immunoglobulin variants outside the Fc region
US20080248028A1 (en) * 2004-03-24 2008-10-09 Xencor, Inc. Immunoglobulin Variants Outside the Fc Region
US20110064727A9 (en) * 2004-03-24 2011-03-17 Xencor, Inc. Immunoglobulin Variants Outside the Fc Region
US7276585B2 (en) 2004-03-24 2007-10-02 Xencor, Inc. Immunoglobulin variants outside the Fc region
US11820830B2 (en) 2004-07-20 2023-11-21 Xencor, Inc. Optimized Fc variants
US20060074225A1 (en) * 2004-09-14 2006-04-06 Xencor, Inc. Monomeric immunoglobulin Fc domains
US8101720B2 (en) 2004-10-21 2012-01-24 Xencor, Inc. Immunoglobulin insertions, deletions and substitutions
US20060173170A1 (en) * 2004-11-12 2006-08-03 Xencor, Inc. Fc variants with altered binding to FcRn
US9803023B2 (en) 2004-11-12 2017-10-31 Xencor, Inc. Fc variants with altered binding to FcRn
US8318907B2 (en) 2004-11-12 2012-11-27 Xencor, Inc. Fc variants with altered binding to FcRn
US11198739B2 (en) 2004-11-12 2021-12-14 Xencor, Inc. Fc variants with altered binding to FcRn
US8546543B2 (en) 2004-11-12 2013-10-01 Xencor, Inc. Fc variants that extend antibody half-life
US8324351B2 (en) 2004-11-12 2012-12-04 Xencor, Inc. Fc variants with altered binding to FcRn
US8852586B2 (en) 2004-11-12 2014-10-07 Xencor, Inc. Fc variants with altered binding to FcRn
US8367805B2 (en) 2004-11-12 2013-02-05 Xencor, Inc. Fc variants with altered binding to FcRn
US8883973B2 (en) 2004-11-12 2014-11-11 Xencor, Inc. Fc variants with altered binding to FcRn
US10336818B2 (en) 2004-11-12 2019-07-02 Xencor, Inc. Fc variants with altered binding to FcRn
US8802820B2 (en) 2004-11-12 2014-08-12 Xencor, Inc. Fc variants with altered binding to FcRn
US8338574B2 (en) 2004-11-12 2012-12-25 Xencor, Inc. FC variants with altered binding to FCRN
US9200079B2 (en) 2004-11-12 2015-12-01 Xencor, Inc. Fc variants with altered binding to FcRn
US20060275282A1 (en) * 2005-01-12 2006-12-07 Xencor, Inc. Antibodies and Fc fusion proteins with altered immunogenicity
US10307481B2 (en) 2005-07-25 2019-06-04 Aptevo Research And Development Llc CD37 immunotherapeutics and uses thereof
US10143748B2 (en) 2005-07-25 2018-12-04 Aptevo Research And Development Llc B-cell reduction using CD37-specific and CD20-specific binding molecules
US20100249382A1 (en) * 2005-10-03 2010-09-30 Xencor, Inc. MODIFIED Fc MOLECULES
US9040041B2 (en) 2005-10-03 2015-05-26 Xencor, Inc. Modified FC molecules
US20080206867A1 (en) * 2005-10-03 2008-08-28 Desjarlais John R Fc variants with optimized Fc receptor binding properties
US7973136B2 (en) 2005-10-06 2011-07-05 Xencor, Inc. Optimized anti-CD30 antibodies
US9574006B2 (en) 2005-10-06 2017-02-21 Xencor, Inc. Optimized anti-CD30 antibodies
US20080267976A1 (en) * 2005-10-06 2008-10-30 Gregory Alan Lazar Optimized Anti-Cd30 Antibodies
US8409577B2 (en) 2006-06-12 2013-04-02 Emergent Product Development Seattle, Llc Single chain multivalent binding proteins with effector function
US9803020B2 (en) 2006-08-14 2017-10-31 Xencor, Inc. Optimized antibodies that target CD19
US11618788B2 (en) 2006-08-14 2023-04-04 Xencor, Inc. Optimized antibodies that target CD19
US20100272723A1 (en) * 2006-08-14 2010-10-28 Xencor, Inc. Optimized Antibodies that Target CD19
US8524867B2 (en) 2006-08-14 2013-09-03 Xencor, Inc. Optimized antibodies that target CD19
US10626182B2 (en) 2006-08-14 2020-04-21 Xencor, Inc. Optimized antibodies that target CD19
US9040042B2 (en) 2006-09-18 2015-05-26 Xencor, Inc. Optimized antibodies that target HM1.24
US8394374B2 (en) 2006-09-18 2013-03-12 Xencor, Inc. Optimized antibodies that target HM1.24
US20100104557A1 (en) * 2006-09-18 2010-04-29 Xencor, Inc. Optimized Antibodies that Target HM1.24
US8629245B2 (en) 2007-05-01 2014-01-14 Research Development Foundation Immunoglobulin Fc libraries
US10725037B2 (en) 2007-05-01 2020-07-28 Research Development Foundation Immunoglobulin Fc libraries
US20090136936A1 (en) * 2007-05-01 2009-05-28 George Georgiou Immunoglobulin fc libraries
US20080313379A1 (en) * 2007-06-15 2008-12-18 United Memories, Inc. Multiple bus charge sharing
US11932685B2 (en) 2007-10-31 2024-03-19 Xencor, Inc. Fc variants with altered binding to FcRn
US9101609B2 (en) 2008-04-11 2015-08-11 Emergent Product Development Seattle, Llc CD37 immunotherapeutic and combination with bifunctional chemotherapeutic thereof
US9879249B2 (en) 2009-02-17 2018-01-30 Redwood Bioscience, Inc. Aldehyde-tagged protein-based drug carriers and methods of use
US9238878B2 (en) 2009-02-17 2016-01-19 Redwood Bioscience, Inc. Aldehyde-tagged protein-based drug carriers and methods of use
US20100330076A1 (en) * 2009-06-30 2010-12-30 George Georgiou Immunoglobulin fc polypeptides
US8679493B2 (en) 2009-06-30 2014-03-25 Research Development Foundation Immunoglobulin Fc polypeptides
US11401348B2 (en) 2009-09-02 2022-08-02 Xencor, Inc. Heterodimeric Fc variants
US9475881B2 (en) 2010-01-19 2016-10-25 Xencor, Inc. Antibody variants with enhanced complement activity
US9540438B2 (en) 2011-01-14 2017-01-10 Redwood Bioscience, Inc. Aldehyde-tagged immunoglobulin polypeptides and methods of use thereof
US10183998B2 (en) 2011-01-14 2019-01-22 Redwood Bioscience, Inc. Aldehyde-tagged immunoglobulin polypeptides and methods of use thereof
US8969526B2 (en) 2011-03-29 2015-03-03 Roche Glycart Ag Antibody Fc variants
US10654916B2 (en) 2011-04-21 2020-05-19 The Regents Of The University Of California, A California Corporation Compositions and methods for the treatment of neuromyelitis optica
US11390667B2 (en) 2011-04-21 2022-07-19 The Regents Of The University Of California Compositions and methods for the treatment of neuromyelitis optica
US10683345B2 (en) 2012-07-13 2020-06-16 Roche Glycart Ag Bispecific anti-VEGF/anti-ANG-2 antibodies and their use in the treatment of ocular vascular diseases
US9695233B2 (en) 2012-07-13 2017-07-04 Roche Glycart Ag Bispecific anti-VEGF/anti-ANG-2 antibodies and their use in the treatment of ocular vascular diseases
US11332538B2 (en) 2015-02-09 2022-05-17 Research Development Foundation Engineered immunoglobulin Fc polypeptides displaying improved complement activation
US10457737B2 (en) 2015-02-09 2019-10-29 Research Development Foundation Engineered immunoglobulin Fc polypeptides displaying improved complement activation
US11352426B2 (en) 2015-09-21 2022-06-07 Aptevo Research And Development Llc CD3 binding polypeptides
US11208632B2 (en) 2016-04-26 2021-12-28 R.P. Scherer Technologies, Llc Antibody conjugates and methods of making and using the same
US11788066B2 (en) 2016-04-26 2023-10-17 R.P. Scherer Technologies, Llc Antibody conjugates and methods of making and using the same

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