WO2023085978A1 - Monoclonal antibody or antigen-binding fragment thereof that specifically binds to il-4ra, and use thereof - Google Patents

Monoclonal antibody or antigen-binding fragment thereof that specifically binds to il-4ra, and use thereof Download PDF

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WO2023085978A1
WO2023085978A1 PCT/RU2022/050351 RU2022050351W WO2023085978A1 WO 2023085978 A1 WO2023085978 A1 WO 2023085978A1 RU 2022050351 W RU2022050351 W RU 2022050351W WO 2023085978 A1 WO2023085978 A1 WO 2023085978A1
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seq
amino acid
acid sequence
variable domain
chain variable
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PCT/RU2022/050351
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French (fr)
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Alina Valerevna BELIASNIKOVA
Olga Leonidovna KYTMANOVA
Darya Olegovna CHERNYSHOVA
Mariia Aleksandrovna SHCHEMELEVA
Anastasiia Isaevna AFREMOVA
Dmitry Valentinovich MOROZOV
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Joint Stock Company «Biocad»
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Priority claimed from RU2021132869A external-priority patent/RU2808563C2/en
Application filed by Joint Stock Company «Biocad» filed Critical Joint Stock Company «Biocad»
Publication of WO2023085978A1 publication Critical patent/WO2023085978A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/715—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons
    • C07K14/7155—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons for interleukins [IL]
    • 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
    • C07K16/2866—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for cytokines, lymphokines, interferons
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/70—Vectors or expression systems specially adapted for E. coli
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/52—Cytokines; Lymphokines; Interferons
    • C07K14/54—Interleukins [IL]
    • C07K14/5406—IL-4
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/33—Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/52—Constant or Fc region; Isotype
    • C07K2317/53—Hinge
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/565—Complementarity determining region [CDR]
    • 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
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Definitions

  • the present invention relates to the field of biotechnology and medicine, in particular to a monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4R ⁇ (interleukin-4 receptor subunit alpha).
  • the invention further relates to nucleic acids encoding said antibody, expression vectors, host cells and methods for producing same, methods for producing the antibodies according to the invention, pharmaceutical compositions comprising the antibody according to the invention, pharmaceutical compositions comprising the antibody according to the invention and other therapeutically active compounds, methods for treating diseases or disorders mediated by IL-4R ⁇ , uses of the antibodies or pharmaceutical compositions thereof for treating diseases or disorders mediated by IL-4R ⁇ , and uses of the antibodies and other therapeutically active compounds for treating diseases or disorders mediated by IL-4R ⁇ .
  • Interleukin 4 is a proinflammatory cytokine mainly produced by T-lymphocytes of the Th2 subpopulation.
  • IL-4 induces the synthesis of cytokines and chemokines, including IL-4 itself and IL-5, IL-9, IL-13, eotaxin and CCL17.
  • IL-4 (together with IL-13) regulates the B cell switching to produce immunoglobulin E.
  • the wide range of effects exerted by IL-4 on various cells is mediated by two variants of the IL-4 receptor (IL-4R), which consist of three types of subunits: IL-4R ⁇ , IL-2Ry and IL-13R ⁇ .
  • the IL-2Ry subunit, or ⁇ c protein is a common component for IL-2, IL-4, IL-7 receptors and some other receptors.
  • IL-4R ⁇ and ⁇ c form IL-4R, a type I high affinity receptor, expressed on the surface of a wide range of immunocompetent and hematopoietic cells as follows: T- and B -lymphocytes, monocytes, granulocytes.
  • the type II receptor for IL-4, IL-4R II is formed by the IL-13R ⁇ and IL-4R ⁇ subunits.
  • IL-4R II is a low affinity receptor capable of binding both IL-4 and IL-13. This phenomenon is contemplated to provide a known overlap in the effects of these cytokines: IL-4 and IL-13, interacting with a receptor, are involved in the development of Th2-dependent inflammation.
  • IL-4 The binding of IL-4 to its receptor takes two steps as follows: first, IL-4 is recruited by the highly affined IL-4R ⁇ ; second, either one of the two less affined subunits (IL-13R ⁇ l or ⁇ C) is recruited into the resulting complex.
  • IL-13 the order of the binding is reversed.
  • IL-13 binds first to the IL-13R ⁇ l with relatively low affinity; second, IL-4R ⁇ , having marginally higher affinity, is recruited into the complex.
  • IL-4R ⁇ In case of blockade of IL-4R ⁇ , IL-4 is not capable of binding to its receptor, and IL- 13, having bound to IL-13R ⁇ l with low affinity, cannot complete the formation of the complex.
  • IL-4 has been shown to have an important pathogenetic role in the formation of bronchial asthma (BA), in particular in the formation of allergic inflammation due to T- lymphocyte proliferation, increased production of immunoglobulin E and eosinophil chemotaxis.
  • BA bronchial asthma
  • IL-4 and IL-13 has been shown to play an important role in pathogenesis as they are involved in Th2 inflammation. It is expected that binding of the IL-4R receptor will result in increased level of these interleukins in the blood.
  • the blockade of IL-4R ⁇ will disturb the formation of Th2 inflammation, thus effecting the clinical manifestations of the above diseases and significantly improving the quality of life of patients with severe chronic pathologies.
  • the international application W02008054606 provides the antibody dupilumab that specifically binds to IL-4R ⁇ .
  • Figure l is a map of an expression vector bearing the genetic sequence of variable domains.
  • Figure 2 is a map of vector bearing the genetic sequence of the antibody heavy chain.
  • Figure 3 is a map of vector bearing the genetic sequence of the antibody light chain. With respect to Figures 2-3
  • Figure 4 is an electrophoregram of antibodies 01 - 08 in 7.5 % polyacrylamide gel under denaturing non-reducing conditions following the first stage of purification on Protein A sorbent.
  • M is molecular weight marker
  • Figure 5 is an electrophoregram of antibodies 09 - 16 in 7.5 % polyacrylamide gel under denaturing non-reducing conditions following the first stage of purification on Protein A sorbent.
  • M is molecular weight marker
  • Figure 6 is an electrophoregram of antibody 17 - 22 in 7.5 % polyacrylamide gel under denaturing non-reducing conditions following the first stage of purification on Protein A sorbent.
  • M is molecular weight marker
  • Figure 7 is an electrophoregram of antibodies 23 - 29 in 7.5 % polyacrylamide gel under denaturing non-reducing conditions following the first stage of purification on Protein A sorbent.
  • M is molecular weight marker
  • Figure 8 is an electrophoregram of antibodies 30 - 037 in 7.5 % polyacrylamide gel under denaturing non-reducing conditions following the first stage of purification on Protein A sorbent.
  • M is molecular weight marker
  • Binding affinity generally refers to the strength of the sum total of noncovalent interactions between a single binding site of a molecule (e.g. an antibody) and its binding partner (e.g. an antigen). Unless indicated otherwise, "binding affinity” refers to intrinsic (characteristic, true) binding affinity which reflects a 1 : 1 interaction between members of a binding pair (e.g. antibody and antigen).
  • the affinity of a molecule X for its binding partner Y can generally be represented by the affinity constant (KD).
  • the preferred Kd value is about 200 nM, 150 nM, 100 nM, 60 nM, 50 nM, 40 nM, 30 nM, 20 nM, 10 nM, 8 nM, 6 nM, 4 nM, 2 nM, 1 nM, or less.
  • Affinity can be measured by common methods known in the art, including those described in the present description. Low-affinity antibodies generally bind an antigen slowly and tend to dissociate readily, whereas high-affinity antibodies generally bind an antigen faster and tend to remain bound longer. A variety of methods of measuring binding affinity are known in the art, any of which can be used for the purposes of the present invention.
  • Kd refers to the off rate constant of a particular interaction between a binding molecule and antigen.
  • the off rate constant koff can be measured using bio- layer interferometry, for example, using OctetTM system.
  • Ka "kon” or "on-rate” refers to the association rate constant.
  • R 2 refers to the coefficient of determination.
  • Response refers to the antibody-antigen binding signal.
  • in vitro refers to a biological entity, a biological process, or a biological reaction outside the body under artificial conditions.
  • a cell grown in vitro is to be understood as a cell grown in an environment outside the body, e.g. in a test tube, a culture vial, or a microtiter plate.
  • ED 50 (EC 50 ) (50% effective dose/concentration) refers to concentrations of a formulation producing 50% biological effect (which may include cytoxicity).
  • the authors of the present group of inventions have developed antibodies that specifically bind to IL-4R ⁇ and have high affinity parameters for binding to the IL-4R ⁇ antigen.
  • the present invention relates to a monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4R ⁇ (interleukin-4 receptor subunit alpha).
  • IL-4R ⁇ interleukin-4 receptor subunit alpha
  • mAb refers to an antibody that is synthesized and isolated as an individual clonal population of cells.
  • the antibody of the invention is a recombinant antibody.
  • recombinant antibody refers to an antibody that is expressed in a cell or cell line comprising nucleotide sequence(s) encoding antibodies, wherein said nucleotide sequence(s) is (are) not associated with the cell in nature.
  • the present invention relates to an isolated monoclonal antibody or antigen- binding fragment thereof that specifically binds to IL-4R ⁇ and comprises:
  • X 1 T, Y, S, A or H;
  • X 2 M, F, L, I or V;
  • X 4 Y, R or N
  • X 5 S, A or R;
  • X 6 T, Q, E, R or S;
  • X 7 Y, H, G, T or V;
  • X 8 R, V, K or S
  • X 9 Y, M, A, G, L or Q;
  • X 10 D, S, E, G, Q, H or N;
  • X 13 T, R or S;
  • X 14 S, G or D;
  • X 16 V, T or Y;
  • X 19 N, S or T;
  • X 20 Q, E, N or R;
  • X 21 R or H
  • X 25 S or G
  • isolated used to describe various antibodies according to this description refers to an antibody which has been identified and isolated and/or regenerated from a cell or cell culture, in which the antibody is expressed.
  • Impurities contaminant components
  • the isolated polypeptide is typically prepared by at least one purification step.
  • antibody or “immunoglobulin” (Ig) as used in the present description includes whole antibodies.
  • antibody refers to a glycoprotein comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds, or antigen-binding portion thereof.
  • Each heavy chain comprises a heavy chain variable region (abbreviated referred to in the present description as VH) and a heavy chain constant region.
  • VH heavy chain variable region
  • the type of a heavy chain present defines the class of an antibody; these chains are found in IgA, IgD, IgE, IgG, and IgM antibodies, respectively.
  • Distinct heavy chains differ in size and composition; ⁇ and ⁇ contain approximately 450 amino acids, while p and a have approximately 550 amino acids.
  • the constant region is identical in all antibodies of the same isotype, but differs in antibodies of different isotypes.
  • Each light chain consists of a light chain variable region (abbreviated referred to in the present description as VL) and light chain constant region.
  • the approximate length of a light chain is 211 to 217 amino acids.
  • the light chain is a kappa (K) light chain
  • the constant domain CL is preferably C kappa (K).
  • Antibodies may be of any class (e.g., IgA, IgD, IgE, IgG, and IgM, preferably IgG), or subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2, preferably IgG4).
  • class e.g., IgA, IgD, IgE, IgG, and IgM, preferably IgG
  • subclass e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2, preferably IgG4
  • VL and VH regions can be further subdivided into hyper-variability regions called complementarity determining regions (CDRs), interspersed between regions that are more conserved, termed framework regions (FR).
  • CDRs complementarity determining regions
  • FR framework regions
  • Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy -terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.
  • the variable regions of heavy and light chains contain a binding domain that interacts with an antigen.
  • the constant regions of antibodies may mediate the binding of immunoglobulin to host tissues or factors, including various cells of the immune system (e.g. effector cells) and the first component (Clq) of the classical complement system.
  • antigen-binding portion of antibody or "antigen -binding fragment”, as used in the present description, refers to one or more antibody fragments that retain the ability to specifically bind to an antigen. It has been shown that the antigen-binding function of antibody can be performed by fragments of a full-length antibody.
  • binding fragments which are included within the term "antigen -binding portion" of an antibody include (i) Fab-fragment, monovalent fragment, consisting of VL, VH, CL and CH1 domains; (ii) F(ab')2 fragment, a bivalent fragment comprising two Fab-fragments linked by a disulfide bridge at the hinge region; (iii) Fd-fragment consisting of VH and CH1 domains; (iv) Fv-fragment consisting of VL and VH domains of a single arm of an antibody; (v) dAb-fragment (Ward et al., (1989) Nature 341:544- 546), which consists of a VH/VHH domain.
  • VL and VH two regions of the Fv-fragment, VL and VH, are encoded by different genes, they can be joined using recombinant methods using a synthetic linker that enables to receive them as a single protein chain in which the VL and VH regions are paired to form monovalent molecules (known as a single-chain Fv (scFv); see e.g. Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879- 5883). It is assumed that such single-stranded molecules are also included within the term "antigen-binding portion" of antibody. Such antibody fragments are produced using conventional techniques known to those skilled in the art, and these fragments are screened in the same manner as intact antibodies are.
  • variable refers to the fact that certain portions of the variable domains greatly differ in sequence among antibodies.
  • the V domain mediates antigen binding and determines specificity of each particular antibody for its particular antigen.
  • variability is not evenly distributed across the 110-amino acid span of the variable domains.
  • the V regions consist of invariant fragments termed framework regions (FRs) of 15-30 amino acids separated by shorter regions of extreme variability termed “hypervariable regions” or CDRs.
  • FRs framework regions
  • hypervariable regions or CDRs.
  • the variable domains of native heavy and light chains each comprise four FRs, largely adopting a beta-sheet configuration, connected by three hypervariable regions, which form loops connecting, and in some cases forming part of, the beta-sheet structure.
  • the hypervariable regions in each chain are held together in close proximity by FRs and, with the hypervariable regions 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 of antibody to antigen, but exhibit various effector functions, such as participation of antibody in antibody- dependent cellular cytotoxicity (ADCC).
  • hypervariable region refers to the amino acid residues of antibody which are responsible for antigen binding.
  • the hypervariable region typically comprises amino acid residues from a “complementarity determining region” or "CDR" and/or those residues from a “hypervariable loop”.
  • Kabat numbering scheme or “numbering according to Kabat” as used in this application refers to the system for numbering of amino acid residues that are more variable (i.e. hypervariable) than other amino acid residues in variable regions of heavy and light chains of the antibody (Kabat et al. Ann. N.Y. Acad. Sci., 190:382-93 (1971); Kabat et al. Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242 (1991)).
  • the antibody of the present invention "which binds" a target antigen refers to an antibody that binds the antigen with sufficient affinity such that the antibody can be used as a diagnostic and/or therapeutic agent targeting a protein or cell or tissue expressing the antigen, and slightly cross-reacts with other proteins.
  • analytical methods fluorescence-activated cell sorting (FACS), radioimmunoassay (RIA) or ELISA, in such embodiments, the degree of antibody binding to a non-target protein is less than 10 % of antibody binding to a specific target protein.
  • the term “specific binding” or phrases “specifically binds to” or “is specific for” a particular polypeptide or an epitope on a particular target polypeptide means binding that is significantly (measurably) different from a non-specific interaction.
  • Specific binding may be measured, for example, by determining binding of a molecule as compared to binding of a control molecule. For example, specific binding may be determined by competition with another molecule that is similar to the target, for example, an excess of non- labeled target. In this case, specific binding is indicated if the binding of the labeled target to a probe is competitively inhibited by the excess of unlabeled target.
  • the term “specific binding” or phrases “specifically binds to” or “is specific for” a particular polypeptide or an epitope on a particular target polypeptide may be described by example of a molecule having a Kd for the target of at least about 200 nM, or at least about 150 nM, or at least about 100 nM, or at least about 60 nM, or at least about 50 nM, or at least about 40 nM, or at least about 30 nM, or at least about 20 nM, or at least about 10 nM, or at least about 8 nM, or at least about 6 nM, or at least about 4 nM, or at least about 2 nM, or at least about 1 nM, or greater.
  • the term “specific binding” refers to binding where a molecule binds to a particular polypeptide or epitope on a particular polypeptide without substantially binding to any other polypeptide or epitope on a polypeptide.
  • the isolated monoclonal antibody or antigen- binding fragment thereof includes a heavy chain variable domain that comprises CDR1 with an amino acid sequence that is selected from the group: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10 or SEQ ID NO: 362.
  • the isolated monoclonal antibody or antigen- binding fragment thereof includes a heavy chain variable domain that comprises CDR2 with an amino acid sequence that is selected from the group: SEQ ID NO: 11 or SEQ ID NO: 12.
  • the isolated monoclonal antibody or antigen- binding fragment thereof includes a heavy chain variable domain that comprises CDR3 with an amino acid sequence that is selected from the group: SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30 or SEQ ID NO: 363.
  • the isolated monoclonal antibody or antigen- binding fragment thereof includes a light chain variable domain that comprises CDR1 with an amino acid sequence that is selected from the group: SEQ ID NO: 31, SEQ ID NO: 32 or SEQ ID NO: 33.
  • the isolated monoclonal antibody or antigen- binding fragment thereof includes a light chain variable domain that comprises CDR2 with an amino acid sequence that is selected from the group: SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38 or SEQ ID NO: 39.
  • the isolated monoclonal antibody or antigen- binding fragment thereof includes a light chain variable domain that comprises CDR3 with an amino acid sequence that is selected from the group: SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42 or SEQ ID NO: 43.
  • the isolated monoclonal antibody or antigen- binding fragment thereof includes:
  • CDR1 with an amino acid sequence selected from the group: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10 or SEQ ID NO: 362;
  • CDR2 with an amino acid sequence selected from the group: SEQ ID NO: 11 or SEQ ID NO: 12;
  • CDR3 with an amino acid sequence selected from the group: SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30 or SEQ ID NO: 363; and
  • CDR1 with an amino acid sequence selected from the group: SEQ ID NO: 31, SEQ ID NO: 32 or SEQ ID NO: 33;
  • CDR2 with an amino acid sequence selected from the group: SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38 or SEQ ID NO: 39; and
  • CDR3 with an amino acid sequence selected from the group: SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42 or SEQ ID NO: 43.
  • the isolated monoclonal antibody or antigen- binding fragment thereof includes:
  • CDR1 with the amino acid sequence of SEQ ID NO: 32, CDR2 with the amino acid sequence of SEQ ID NO: 35 and CDR3 with the amino acid sequence of SEQ ID NO: 41;
  • a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 1, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 15; and (b) a light chain variable domain comprising:
  • CDR1 with the amino acid sequence of SEQ ID NO: 32, CDR2 with the amino acid sequence of SEQ ID NO: 35 and CDR3 with the amino acid sequence of SEQ ID NO: 41;
  • CDR1 with the amino acid sequence of SEQ ID NO: 31, CDR2 with the amino acid sequence of SEQ ID NO: 34 and CDR3 with the amino acid sequence of SEQ ID NO: 40;
  • a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 1, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 17; and (b) a light chain variable domain comprising:
  • CDR1 with the amino acid sequence of SEQ ID NO: 31, CDR2 with the amino acid sequence of SEQ ID NO: 34 and CDR3 with the amino acid sequence of SEQ ID NO: 40;
  • a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 2, CDR2 with the amino acid sequence of SEQ ID NO: 12 and CDR3 with the amino acid sequence of SEQ ID NO: 18; and (b) a light chain variable domain comprising:
  • xv (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 8, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 25, and (b) a light chain variable domain comprising:
  • the isolated monoclonal antibody or antigen- binding fragment thereof includes a heavy chain variable domain that comprises an amino acid sequence that is selected from the group: SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72
  • the isolated monoclonal antibody or antigen- binding fragment thereof includes a light chain variable domain that comprises an amino acid sequence that is selected from the group: SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86 or SEQ ID NO: 87.
  • the isolated monoclonal antibody or antigen- binding fragment thereof includes:
  • a heavy chain variable domain that comprises an amino acid sequence selected from the group: SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 366, SEQ ID NO:
  • a light chain variable domain that comprises an amino acid sequence selected from the group: SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86 or SEQ ID NO: 87.
  • the isolated monoclonal antibody or antigen- binding fragment thereof includes:
  • the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 85;
  • the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 53 and
  • the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 86;
  • the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 53;
  • the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 87;
  • the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 368;
  • the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74;
  • the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 370;
  • the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74;
  • the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 372;
  • the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74.
  • the isolated monoclonal antibody that specifically binds to IL-4R ⁇ is a full-length IgG antibody.
  • the isolated monoclonal antibody is a full-length IgG antibody that is of human IgGl, IgG2, IgG3 or IgG4 isotype.
  • the isolated monoclonal antibody is a full-length IgG antibody that is of human IgG4 isotype.
  • the isolated monoclonal antibody comprises the mutation S228P in the hinge region.
  • the above mutation in the hinge region is numbered according to the EU numbering for amino acid chains of antibodies (Edelman, G.M., et al., Proc. Natl. Acad. Sci. USA 63 (1969), pp. 78-85; Kabat, E.A., et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD, (1991).
  • the same mutation can be designated as S241P according to the Kabat numbering scheme for numbering amino acids of antibody chains, or as S229P according to sequential numbering from the beginning of the chain.
  • the isolated monoclonal antibody includes a heavy chain that comprises an amino acid sequence that is selected from the group: SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO:
  • the isolated monoclonal antibody includes a light chain that comprises an amino acid sequence that is selected from the group: SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130 or SEQ ID NO: 131.
  • the isolated monoclonal antibody includes:
  • a light chain comprising an amino acid sequence that is selected from the group: SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130 or SEQ ID NO: 131;
  • the isolated monoclonal antibody that specifically binds to IL-4R ⁇ is antibody 01.
  • Antibody 01 includes:
  • Antibody 01 includes:
  • Antibody 01 includes:
  • Antibody 01 includes:
  • Antibody 01 includes:
  • the isolated monoclonal antibody that specifically binds to IL-4R ⁇ is antibody 02.
  • Antibody 02 includes:
  • Antibody 02 includes:
  • Antibody 02 includes:
  • Antibody 02 includes:
  • Antibody 02 includes:
  • the isolated monoclonal antibody that specifically binds to IL-4R ⁇ is antibody 03.
  • Antibody 03 includes:
  • Antibody 03 includes:
  • Antibody 03 includes:
  • Antibody 03 includes:
  • Antibody 03 includes:
  • CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 204
  • CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 207
  • the isolated monoclonal antibody that specifically binds to IL-4R ⁇ is antibody 04.
  • Antibody 04 includes:
  • Antibody 04 includes:
  • Antibody 04 includes:
  • Antibody 04 includes:
  • Antibody 04 includes:
  • the isolated monoclonal antibody that specifically binds to IL-4R ⁇ is antibody 05.
  • Antibody 05 includes:
  • Antibody 05 includes:
  • Antibody 05 includes:
  • Antibody 05 includes:
  • Antibody 05 includes:
  • CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178
  • CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184
  • the isolated monoclonal antibody that specifically binds to IL-4R ⁇ is antibody 06.
  • Antibody 06 includes:
  • Antibody 06 includes:
  • Antibody 06 includes:
  • Antibody 06 includes:
  • Antibody 06 includes: (a) a heavy chain variable domain comprising:
  • the isolated monoclonal antibody that specifically binds to IL-4R ⁇ is antibody 07.
  • Antibody 07 includes:
  • Antibody 07 includes:
  • Antibody 07 includes:
  • Antibody 07 includes:
  • Antibody 07 includes:
  • the isolated monoclonal antibody that specifically binds to IL-4R ⁇ is antibody 08.
  • Antibody 08 includes:
  • Antibody 08 includes:
  • Antibody 08 includes:
  • Antibody 08 includes:
  • a light chain variable domain comprising: (i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
  • Antibody 08 includes:
  • the isolated monoclonal antibody that specifically binds to IL-4R ⁇ is antibody 09.
  • Antibody 09 includes:
  • Antibody 09 includes:
  • Antibody 09 includes:
  • Antibody 09 includes:
  • Antibody 09 includes:
  • the isolated monoclonal antibody that specifically binds to IL-4R ⁇ is antibody 10.
  • Antibody 10 includes:
  • Antibody 10 includes:
  • Antibody 10 includes:
  • Antibody 10 includes:
  • a heavy chain variable domain comprising: (i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
  • Antibody 10 includes:
  • the isolated monoclonal antibody that specifically binds to IL-4R ⁇ is antibody 11.
  • Antibody 11 includes:
  • Antibody 11 includes:
  • Antibody 11 includes:
  • Antibody 11 includes:
  • Antibody 11 includes:
  • the isolated monoclonal antibody that specifically binds to IL-4R ⁇ is antibody 12.
  • Antibody 12 includes:
  • Antibody 12 includes:
  • Antibody 12 includes:
  • Antibody 12 includes:
  • Antibody 12 includes:
  • the isolated monoclonal antibody that specifically binds to IL-4R ⁇ is antibody 13.
  • Antibody 13 includes:
  • Antibody 13 includes:
  • Antibody 13 includes:
  • Antibody 13 includes:
  • Antibody 13 includes:
  • the isolated monoclonal antibody that specifically binds to IL-4R ⁇ is antibody 14.
  • Antibody 14 includes:
  • Antibody 14 includes:
  • Antibody 14 includes:
  • Antibody 14 includes:
  • Antibody 14 includes:
  • the isolated monoclonal antibody that specifically binds to IL-4R ⁇ is antibody 15.
  • Antibody 15 includes:
  • Antibody 15 includes:
  • Antibody 15 includes: (a) a heavy chain variable domain comprising:
  • Antibody 15 includes:
  • Antibody 15 includes:
  • the isolated monoclonal antibody that specifically binds to IL-4R ⁇ is antibody 16.
  • Antibody 16 includes:
  • Antibody 16 includes:
  • Antibody 16 includes:
  • Antibody 16 includes:
  • Antibody 16 includes:
  • the isolated monoclonal antibody that specifically binds to IL-4R ⁇ is antibody 17.
  • Antibody 17 includes:
  • Antibody 17 includes: (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO:
  • Antibody 17 includes:
  • Antibody 17 includes:
  • Antibody 17 includes:
  • the isolated monoclonal antibody that specifically binds to IL-4R ⁇ is antibody 18.
  • Antibody 18 includes:
  • Antibody 18 includes:
  • Antibody 18 includes:

Abstract

The present invention relates to the field of biotechnology and medicine, in particular to a monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα (interleukin-4 receptor subunit alpha). The invention further relates to nucleic acids encoding said antibody, expression vectors, host cells and methods for producing same, methods for producing the antibodies according to the invention, pharmaceutical compositions comprising the antibody according to the invention, pharmaceutical compositions comprising the antibody according to the invention and other therapeutically active compounds, methods for treating diseases or disorders mediated by IL-4Rα, uses of the antibodies or pharmaceutical compositions thereof for treating diseases or disorders mediated by IL-4Rα, and uses of the antibodies and other therapeutically active compounds for treating diseases or disorders mediated by IL-4Rα.

Description

MONOCLONAL ANTIBODY OR ANTIGEN-BINDING FRAGMENT THEREOF THAT
SPECIFICALLY BINDS TO IL-4Rα, AND USE THEREOF
Field of the invention
The present invention relates to the field of biotechnology and medicine, in particular to a monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα (interleukin-4 receptor subunit alpha). The invention further relates to nucleic acids encoding said antibody, expression vectors, host cells and methods for producing same, methods for producing the antibodies according to the invention, pharmaceutical compositions comprising the antibody according to the invention, pharmaceutical compositions comprising the antibody according to the invention and other therapeutically active compounds, methods for treating diseases or disorders mediated by IL-4Rα, uses of the antibodies or pharmaceutical compositions thereof for treating diseases or disorders mediated by IL-4Rα, and uses of the antibodies and other therapeutically active compounds for treating diseases or disorders mediated by IL-4Rα.
Background of the invention
Interleukin 4 (IL-4) is a proinflammatory cytokine mainly produced by T-lymphocytes of the Th2 subpopulation. In addition to Th2 cell differentiation, IL-4 induces the synthesis of cytokines and chemokines, including IL-4 itself and IL-5, IL-9, IL-13, eotaxin and CCL17. Furthermore, IL-4 (together with IL-13) regulates the B cell switching to produce immunoglobulin E. The wide range of effects exerted by IL-4 on various cells is mediated by two variants of the IL-4 receptor (IL-4R), which consist of three types of subunits: IL-4Rα, IL-2Ry and IL-13Rα.
The IL-2Ry subunit, or γc protein, is a common component for IL-2, IL-4, IL-7 receptors and some other receptors. When combined, IL-4Rα and γc form IL-4R, a type I high affinity receptor, expressed on the surface of a wide range of immunocompetent and hematopoietic cells as follows: T- and B -lymphocytes, monocytes, granulocytes. The type II receptor for IL-4, IL-4R II, is formed by the IL-13Rα and IL-4Rα subunits. IL-4R II is a low affinity receptor capable of binding both IL-4 and IL-13. This phenomenon is contemplated to provide a known overlap in the effects of these cytokines: IL-4 and IL-13, interacting with a receptor, are involved in the development of Th2-dependent inflammation.
The binding of IL-4 to its receptor takes two steps as follows: first, IL-4 is recruited by the highly affined IL-4Rα; second, either one of the two less affined subunits (IL-13Rαl or γC) is recruited into the resulting complex. For IL-13, the order of the binding is reversed. IL-13 binds first to the IL-13Rαl with relatively low affinity; second, IL-4Rα, having marginally higher affinity, is recruited into the complex. Thus, in case of blockade of IL-4Rα, IL-4 is not capable of binding to its receptor, and IL- 13, having bound to IL-13Rαl with low affinity, cannot complete the formation of the complex.
To date, IL-4 has been shown to have an important pathogenetic role in the formation of bronchial asthma (BA), in particular in the formation of allergic inflammation due to T- lymphocyte proliferation, increased production of immunoglobulin E and eosinophil chemotaxis.
All patients with bronchial asthma have shown significant fluctuations in IL-4 levels in the peripheral blood: a significant increase in the levels of the cytokine has been observed in patients with any severity of BA during the disease exacerbation, with a tendency to decrease in the remission phase. Still, the concentration of IL-4 in the blood of BA patients (including those with mild course) always exceeded that in healthy individuals.
For atopic dermatitis, IL-4 and IL-13 has been shown to play an important role in pathogenesis as they are involved in Th2 inflammation. It is expected that binding of the IL-4R receptor will result in increased level of these interleukins in the blood.
Two approaches have been developed to inhibit IL- 13 and/or IL-4 as follows:
1) inhalation of dissolved IL-4R, a selective IL-4 antagonist;
2) blockade of IL-4Rα, a key signaling component of the IL-4 and IL- 13 receptor complex.
Thus, the blockade of IL-4Rα will disturb the formation of Th2 inflammation, thus effecting the clinical manifestations of the above diseases and significantly improving the quality of life of patients with severe chronic pathologies.
The international application W02008054606 provides the antibody dupilumab that specifically binds to IL-4Rα.
To date, only one antibody to IL-4Rα (dupilumab) has been approved for therapeutic use. In connection with the above, there is a need for novel antibodies that specifically bind to IL-4Rα.
Brief description of drawings
Figure l is a map of an expression vector bearing the genetic sequence of variable domains.
Figure imgf000004_0001
Figure imgf000005_0001
Figure 2 is a map of vector bearing the genetic sequence of the antibody heavy chain.
Figure 3 is a map of vector bearing the genetic sequence of the antibody light chain. With respect to Figures 2-3
Figure imgf000005_0002
Figure imgf000006_0001
Figure 4 is an electrophoregram of antibodies 01 - 08 in 7.5 % polyacrylamide gel under denaturing non-reducing conditions following the first stage of purification on Protein A sorbent.
M is molecular weight marker.
1- antibody 01.
2- antibody 02.
3- antibody 03.
4- antibody 04.
5- antibody 05.
6- antibody 06.
7- antibody 07.
8- antibody 08.
Figure 5 is an electrophoregram of antibodies 09 - 16 in 7.5 % polyacrylamide gel under denaturing non-reducing conditions following the first stage of purification on Protein A sorbent.
M is molecular weight marker.
1- antibody 09.
2- antibody 10.
3- antibody 11.
4- antibody 12.
5- antibody 13.
6- antibody 14.
7- antibody 15.
8- antibody 16.
Figure 6 is an electrophoregram of antibody 17 - 22 in 7.5 % polyacrylamide gel under denaturing non-reducing conditions following the first stage of purification on Protein A sorbent.
M is molecular weight marker.
1- antibody 17.
2- antibody 18.
3- antibody 19. 4- antibody 20.
5- antibody 21.
6- antibody 22.
Figure 7 is an electrophoregram of antibodies 23 - 29 in 7.5 % polyacrylamide gel under denaturing non-reducing conditions following the first stage of purification on Protein A sorbent.
M is molecular weight marker.
1- antibody 23.
2- antibody 24.
3- antibody 25.
4- antibody 26.
5- antibody 27.
6- antibody 28.
7- antibody 29.
Figure 8 is an electrophoregram of antibodies 30 - 037 in 7.5 % polyacrylamide gel under denaturing non-reducing conditions following the first stage of purification on Protein A sorbent.
M is molecular weight marker.
1- antibody 30.
2- antibody 31.
3- antibody 32.
4- antibody 33.
5- antibody 34.
6- antibody 35.
7- antibody 36.
8- antibody 37.
Definitions and general methods
Unless defined otherwise herein, all technical and scientific terms used in connection with the present invention will have the same meaning as is commonly understood by those skilled in the art.
Furthermore, unless otherwise required by context, singular terms shall include plural terms, and the plural terms shall include the singular terms. Typically, the present classification and methods of cell culture, molecular biology, immunology, microbiology, genetics, analytical chemistry, organic synthesis chemistry, medical and pharmaceutical chemistry, as well as hybridization and chemistry of protein and nucleic acids described herein are well known by those skilled and widely used in the art. Enzyme reactions and purification methods are performed according to the manufacturer's guidelines, as is common in the art, or as described herein. The term "KD" in this description refers to the affinity constant (or equilibrium constant), which is calculated from the ratio of Kd to Ka (i.e. Kd/Ka), and it is expressed as a molar concentration (M).
"Binding affinity" generally refers to the strength of the sum total of noncovalent interactions between a single binding site of a molecule (e.g. an antibody) and its binding partner (e.g. an antigen). Unless indicated otherwise, "binding affinity" refers to intrinsic (characteristic, true) binding affinity which reflects a 1 : 1 interaction between members of a binding pair (e.g. antibody and antigen). The affinity of a molecule X for its binding partner Y can generally be represented by the affinity constant (KD). The preferred Kd value is about 200 nM, 150 nM, 100 nM, 60 nM, 50 nM, 40 nM, 30 nM, 20 nM, 10 nM, 8 nM, 6 nM, 4 nM, 2 nM, 1 nM, or less. Affinity can be measured by common methods known in the art, including those described in the present description. Low-affinity antibodies generally bind an antigen slowly and tend to dissociate readily, whereas high-affinity antibodies generally bind an antigen faster and tend to remain bound longer. A variety of methods of measuring binding affinity are known in the art, any of which can be used for the purposes of the present invention.
The term "Kd", "koff1 or "kdis" refers to the off rate constant of a particular interaction between a binding molecule and antigen. The off rate constant koff can be measured using bio- layer interferometry, for example, using Octet™ system.
The term "Ka", "kon" or "on-rate" refers to the association rate constant.
The term "R2" refers to the coefficient of determination.
The term "Response" refers to the antibody-antigen binding signal.
The term "in vitro" refers to a biological entity, a biological process, or a biological reaction outside the body under artificial conditions. For example, a cell grown in vitro is to be understood as a cell grown in an environment outside the body, e.g. in a test tube, a culture vial, or a microtiter plate.
The term "ED50" (EC50) (50% effective dose/concentration) refers to concentrations of a formulation producing 50% biological effect (which may include cytoxicity).
As used in the present description and claims that follow, unless otherwise dictated by the context, the words "include" and "comprise", or variations thereof such as "includes", "including", "comprises", or "comprising", will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers.
Detailed description of the invention
The authors of the present group of inventions have developed antibodies that specifically bind to IL-4Rα and have high affinity parameters for binding to the IL-4Rα antigen.
Antibody The present invention relates to a monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα (interleukin-4 receptor subunit alpha).
The term "monoclonal antibody" or "mAb" refers to an antibody that is synthesized and isolated as an individual clonal population of cells.
The antibody of the invention is a recombinant antibody.
The term "recombinant antibody" refers to an antibody that is expressed in a cell or cell line comprising nucleotide sequence(s) encoding antibodies, wherein said nucleotide sequence(s) is (are) not associated with the cell in nature.
In one aspect, the present invention relates to an isolated monoclonal antibody or antigen- binding fragment thereof that specifically binds to IL-4Rα and comprises:
(a) a heavy chain variable domain comprising:
(i) CDR1 with the amino acid sequence X1YW X2H, where
X1= T, Y, S, A or H;
X2 = M, F, L, I or V;
(ii) CDR2 with the amino acid sequence X3IRPAGSSTYYTDSVKG, where
X3 = T or S; and
(iii) CDR3 with the amino acid sequence GGX4SX5X6WX7X8X9X10X11, where
X4=Y, R or N;
X5= S, A or R;
X6 = T, Q, E, R or S;
X7= Y, H, G, T or V;
X8=R, V, K or S;
X9=Y, M, A, G, L or Q;
X10=D, S, E, G, Q, H or N;
X11=P, Y or S; and
(b) a light chain variable domain comprising:
(i) CDR1 with the amino acid sequence X12GSX13SNIGX14X15X16VG, where
X12 = S or T;
X13 = T, R or S;
X14= S, G or D;
X15 = N or Y;
X16 = V, T or Y;
(ii) CDR2 with the amino acid sequence X17X18X19X20X21X22S, where
X17 = R, I or S; X18 = D or N;
X19 = N, S or T;
X20= Q, E, N or R;
X21= R or H;
X22= P or A; and
(iii) CDR3 with amino acid sequence X23X24WDDX25LX26X27VV
X23=S, A or T;
X24= T, A or S;
X25 = S or G;
X26 = N or S;
X27 = G or 0.
The term "isolated" used to describe various antibodies according to this description refers to an antibody which has been identified and isolated and/or regenerated from a cell or cell culture, in which the antibody is expressed. Impurities (contaminant components) from natural environment are materials which typically interfere with diagnostic or therapeutic uses of the polypeptide, and may include enzymes, hormones, and other proteinaceous or nonproteinaceous solutes. The isolated polypeptide is typically prepared by at least one purification step.
The term "antibody" or "immunoglobulin" (Ig) as used in the present description includes whole antibodies. The term "antibody" refers to a glycoprotein comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds, or antigen-binding portion thereof. Each heavy chain comprises a heavy chain variable region (abbreviated referred to in the present description as VH) and a heavy chain constant region. Known are five types of mammalian antibody heavy chains denoted by Greek letters: α,
Figure imgf000010_0002
, ε, y and p. (Janeway C.A., Jr. et al, Immunobiology, 5th ed., publ. by Garland Publishing, 2001). The type of a heavy chain present defines the class of an antibody; these chains are found in IgA, IgD, IgE, IgG, and IgM antibodies, respectively. (Rhoades R.A., Pflanzer R.G., Human Physiology, 4th ed., publ. by Thomson Learning, 2002). Distinct heavy chains differ in size and composition; α and γ contain approximately 450 amino acids, while p and a have approximately 550 amino acids. The constant region is identical in all antibodies of the same isotype, but differs in antibodies of different isotypes. Heavy chains y, a and
Figure imgf000010_0001
have a constant region composed of three constant domains CH1, CH2 and CH3 (in a line), and a hinge region for added flexibility (Woof J., Burton D., Nat Rev Immunol 4, 2004, cc.89-99); heavy chains p and a have a constant region composed of four constant domains CH1, CH2, CH3 and CH4 (Janeway C.A., Jr. et al, Immunobiology, 5th ed., publ. by Garland Publishing, 2001). In mammals, known are only two types of light chains denoted by lambda (X) and kappa (K). Each light chain consists of a light chain variable region (abbreviated referred to in the present description as VL) and light chain constant region. The approximate length of a light chain is 211 to 217 amino acids. Preferably the light chain is a kappa (K) light chain, and the constant domain CL is preferably C kappa (K).
"Antibodies" according to the invention may be of any class (e.g., IgA, IgD, IgE, IgG, and IgM, preferably IgG), or subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2, preferably IgG4).
VL and VH regions can be further subdivided into hyper-variability regions called complementarity determining regions (CDRs), interspersed between regions that are more conserved, termed framework regions (FR). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy -terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of heavy and light chains contain a binding domain that interacts with an antigen. The constant regions of antibodies may mediate the binding of immunoglobulin to host tissues or factors, including various cells of the immune system (e.g. effector cells) and the first component (Clq) of the classical complement system.
The term "antigen-binding portion" of antibody or "antigen -binding fragment", as used in the present description, refers to one or more antibody fragments that retain the ability to specifically bind to an antigen. It has been shown that the antigen-binding function of antibody can be performed by fragments of a full-length antibody. Examples of binding fragments which are included within the term "antigen -binding portion" of an antibody include (i) Fab-fragment, monovalent fragment, consisting of VL, VH, CL and CH1 domains; (ii) F(ab')2 fragment, a bivalent fragment comprising two Fab-fragments linked by a disulfide bridge at the hinge region; (iii) Fd-fragment consisting of VH and CH1 domains; (iv) Fv-fragment consisting of VL and VH domains of a single arm of an antibody; (v) dAb-fragment (Ward et al., (1989) Nature 341:544- 546), which consists of a VH/VHH domain. In addition, two regions of the Fv-fragment, VL and VH, are encoded by different genes, they can be joined using recombinant methods using a synthetic linker that enables to receive them as a single protein chain in which the VL and VH regions are paired to form monovalent molecules (known as a single-chain Fv (scFv); see e.g. Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879- 5883). It is assumed that such single-stranded molecules are also included within the term "antigen-binding portion" of antibody. Such antibody fragments are produced using conventional techniques known to those skilled in the art, and these fragments are screened in the same manner as intact antibodies are.
The term "variable" refers to the fact that certain portions of the variable domains greatly differ in sequence among antibodies. The V domain mediates antigen binding and determines specificity of each particular antibody for its particular antigen. However, the variability is not evenly distributed across the 110-amino acid span of the variable domains. Instead, the V regions consist of invariant fragments termed framework regions (FRs) of 15-30 amino acids separated by shorter regions of extreme variability termed "hypervariable regions" or CDRs. The variable domains of native heavy and light chains each comprise four FRs, largely adopting a beta-sheet configuration, connected by three hypervariable regions, which form loops connecting, and in some cases forming part of, the beta-sheet structure. The hypervariable regions in each chain are held together in close proximity by FRs and, with the hypervariable regions 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 of antibody to antigen, but exhibit various effector functions, such as participation of antibody in antibody- dependent cellular cytotoxicity (ADCC).
The term "hypervariable region" according to the present description refers to the amino acid residues of antibody which are responsible for antigen binding. The hypervariable region typically comprises amino acid residues from a "complementarity determining region" or "CDR" and/or those residues from a "hypervariable loop".
“Kabat numbering scheme” or “numbering according to Kabat” as used in this application refers to the system for numbering of amino acid residues that are more variable (i.e. hypervariable) than other amino acid residues in variable regions of heavy and light chains of the antibody (Kabat et al. Ann. N.Y. Acad. Sci., 190:382-93 (1971); Kabat et al. Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242 (1991)).
The antibody of the present invention "which binds" a target antigen refers to an antibody that binds the antigen with sufficient affinity such that the antibody can be used as a diagnostic and/or therapeutic agent targeting a protein or cell or tissue expressing the antigen, and slightly cross-reacts with other proteins. According to analytical methods: fluorescence-activated cell sorting (FACS), radioimmunoassay (RIA) or ELISA, in such embodiments, the degree of antibody binding to a non-target protein is less than 10 % of antibody binding to a specific target protein. With regard to the binding of antibody to a target molecule, the term “specific binding” or phrases “specifically binds to” or “is specific for” a particular polypeptide or an epitope on a particular target polypeptide means binding that is significantly (measurably) different from a non-specific interaction.
Specific binding may be measured, for example, by determining binding of a molecule as compared to binding of a control molecule. For example, specific binding may be determined by competition with another molecule that is similar to the target, for example, an excess of non- labeled target. In this case, specific binding is indicated if the binding of the labeled target to a probe is competitively inhibited by the excess of unlabeled target. As used in the present description, the term "specific binding" or phrases "specifically binds to" or "is specific for" a particular polypeptide or an epitope on a particular target polypeptide may be described by example of a molecule having a Kd for the target of at least about 200 nM, or at least about 150 nM, or at least about 100 nM, or at least about 60 nM, or at least about 50 nM, or at least about 40 nM, or at least about 30 nM, or at least about 20 nM, or at least about 10 nM, or at least about 8 nM, or at least about 6 nM, or at least about 4 nM, or at least about 2 nM, or at least about 1 nM, or greater. In one embodiment, the term "specific binding" refers to binding where a molecule binds to a particular polypeptide or epitope on a particular polypeptide without substantially binding to any other polypeptide or epitope on a polypeptide.
In some embodiments of the invention, the isolated monoclonal antibody or antigen- binding fragment thereof includes a heavy chain variable domain that comprises CDR1 with an amino acid sequence that is selected from the group: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10 or SEQ ID NO: 362.
In some embodiments of the invention, the isolated monoclonal antibody or antigen- binding fragment thereof includes a heavy chain variable domain that comprises CDR2 with an amino acid sequence that is selected from the group: SEQ ID NO: 11 or SEQ ID NO: 12.
In some embodiments of the invention, the isolated monoclonal antibody or antigen- binding fragment thereof includes a heavy chain variable domain that comprises CDR3 with an amino acid sequence that is selected from the group: SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30 or SEQ ID NO: 363.
In some embodiments of the invention, the isolated monoclonal antibody or antigen- binding fragment thereof includes a light chain variable domain that comprises CDR1 with an amino acid sequence that is selected from the group: SEQ ID NO: 31, SEQ ID NO: 32 or SEQ ID NO: 33.
In some embodiments of the invention, the isolated monoclonal antibody or antigen- binding fragment thereof includes a light chain variable domain that comprises CDR2 with an amino acid sequence that is selected from the group: SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38 or SEQ ID NO: 39.
In some embodiments of the invention, the isolated monoclonal antibody or antigen- binding fragment thereof includes a light chain variable domain that comprises CDR3 with an amino acid sequence that is selected from the group: SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42 or SEQ ID NO: 43.
In some embodiments of the invention, the isolated monoclonal antibody or antigen- binding fragment thereof includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 with an amino acid sequence selected from the group: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10 or SEQ ID NO: 362;
(ii) CDR2 with an amino acid sequence selected from the group: SEQ ID NO: 11 or SEQ ID NO: 12; and
(iii) CDR3 with an amino acid sequence selected from the group: SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30 or SEQ ID NO: 363; and
(b) a light chain variable domain comprising:
(i) CDR1 with an amino acid sequence selected from the group: SEQ ID NO: 31, SEQ ID NO: 32 or SEQ ID NO: 33;
(ii) CDR2 with an amino acid sequence selected from the group: SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38 or SEQ ID NO: 39; and
(iii) CDR3 with an amino acid sequence selected from the group: SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42 or SEQ ID NO: 43.
In some embodiments of the invention, the isolated monoclonal antibody or antigen- binding fragment thereof includes:
(i) (a) a heavy chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 1,
CDR2 with the amino acid sequence of SEQ ID NO: 11 and
CDR3 with the amino acid sequence of SEQ ID NO: 13; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 31,
CDR2 with the amino acid sequence of SEQ ID NO: 34 and
CDR3 with the amino acid sequence of SEQ ID NO: 40;
(ii) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 1, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 14; and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 32, CDR2 with the amino acid sequence of SEQ ID NO: 35 and CDR3 with the amino acid sequence of SEQ ID NO: 41;
(iii) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 1, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 15; and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 32, CDR2 with the amino acid sequence of SEQ ID NO: 35 and CDR3 with the amino acid sequence of SEQ ID NO: 41;
(iv) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 1, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 16; and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 31, CDR2 with the amino acid sequence of SEQ ID NO: 34 and CDR3 with the amino acid sequence of SEQ ID NO: 40;
(v) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 1, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 17; and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 31, CDR2 with the amino acid sequence of SEQ ID NO: 34 and CDR3 with the amino acid sequence of SEQ ID NO: 40;
(vi) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 2, CDR2 with the amino acid sequence of SEQ ID NO: 12 and CDR3 with the amino acid sequence of SEQ ID NO: 18; and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 33,
CDR2 with the amino acid sequence of SEQ ID NO: 36 and
CDR3 with the amino acid sequence of SEQ ID NO: 42;
(vii) (a) a heavy chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 3,
CDR2 with the amino acid sequence of SEQ ID NO: 11 and
CDR3 with the amino acid sequence of SEQ ID NO: 19; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 33,
CDR2 with the amino acid sequence of SEQ ID NO: 37 and
CDR3 with the amino acid sequence of SEQ ID NO: 42;
(viii) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 4,
CDR2 with the amino acid sequence of SEQ ID NO: 11 and
CDR3 with the amino acid sequence of SEQ ID NO: 20, and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 33,
CDR2 with the amino acid sequence of SEQ ID NO: 36 and
CDR3 with the amino acid sequence of SEQ ID NO: 42;
(ix) (a) a heavy chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 5,
CDR2 with the amino acid sequence of SEQ ID NO: 11 and
CDR3 with the amino acid sequence of SEQ ID NO: 21; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 33,
CDR2 with the amino acid sequence of SEQ ID NO: 36 and
CDR3 with the amino acid sequence of SEQ ID NO: 42;
(x) (a) a heavy chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 1,
CDR2 with the amino acid sequence of SEQ ID NO: 11 and
CDR3 with the amino acid sequence of SEQ ID NO: 22; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 33,
CDR2 with the amino acid sequence of SEQ ID NO: 36 and CDR3 with the amino acid sequence of SEQ ID NO: 42;
(xi) (a) a heavy chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 6,
CDR2 with the amino acid sequence of SEQ ID NO: 11 and
CDR3 with the amino acid sequence of SEQ ID NO: 23; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 33,
CDR2 with the amino acid sequence of SEQ ID NO: 36 and
CDR3 with the amino acid sequence of SEQ ID NO: 42;
(xii) (a) a heavy chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 1,
CDR2 with the amino acid sequence of SEQ ID NO: 11 and
CDR3 with the amino acid sequence of SEQ ID NO: 17; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 31,
CDR2 with the amino acid sequence of SEQ ID NO: 38 and
CDR3 with the amino acid sequence of SEQ ID NO: 40;
(xiii)(a) a heavy chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 362,
CDR2 with the amino acid sequence of SEQ ID NO: 11 and
CDR3 with the amino acid sequence of SEQ ID NO: 363; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 32,
CDR2 with the amino acid sequence of SEQ ID NO: 35 and
CDR3 with the amino acid sequence of SEQ ID NO: 41;
(xiv) (a) a heavy chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 7,
CDR2 with the amino acid sequence of SEQ ID NO: 11 and
CDR3 with the amino acid sequence of SEQ ID NO: 24; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 32,
CDR2 with the amino acid sequence of SEQ ID NO: 35 and
CDR3 with the amino acid sequence of SEQ ID NO: 41;
(xv) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 8, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 25, and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 32,
CDR2 with the amino acid sequence of SEQ ID NO: 35 and
CDR3 with the amino acid sequence of SEQ ID NO: 41;
(xvi) (a) a heavy chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 9,
CDR2 with the amino acid sequence of SEQ ID NO: 11 and
CDR3 with the amino acid sequence of SEQ ID NO: 21; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 32,
CDR2 with the amino acid sequence of SEQ ID NO: 35 and
CDR3 with the amino acid sequence of SEQ ID NO: 41;
(xvii) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 8,
CDR2 with the amino acid sequence of SEQ ID NO: 11 and
CDR3 with the amino acid sequence of SEQ ID NO: 26; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 32,
CDR2 with the amino acid sequence of SEQ ID NO: 35 and
CDR3 with the amino acid sequence of SEQ ID NO: 41;
(xviii) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 8,
CDR2 with the amino acid sequence of SEQ ID NO: 11 and
CDR3 with the amino acid sequence of SEQ ID NO: 27; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 32,
CDR2 with the amino acid sequence of SEQ ID NO: 35 and
CDR3 with the amino acid sequence of SEQ ID NO: 41;
(xix) (a) a heavy chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 1,
CDR2 with the amino acid sequence of SEQ ID NO: 11 and
CDR3 with the amino acid sequence of SEQ ID NO: 29; and
(b) a light chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 31,
CDR2 with the amino acid sequence of SEQ ID NO: 34 and
CDR3 with the amino acid sequence of SEQ ID NO: 40;
(xx) (a) a heavy chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 8,
CDR2 with the amino acid sequence of SEQ ID NO: 11 and
CDR3 with the amino acid sequence of SEQ ID NO: 30; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 32;
CDR2 with the amino acid sequence of SEQ ID NO: 35 and
CDR3 with the amino acid sequence of SEQ ID NO: 41;
(xxi) CDR1 with the amino acid sequence of SEQ ID NO: 1,
CDR2 with the amino acid sequence of SEQ ID NO: 11 and
CDR3 with the amino acid sequence of SEQ ID NO: 17; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 31,
CDR2 with the amino acid sequence of SEQ ID NO: 34 and
CDR3 with the amino acid sequence of SEQ ID NO: 43; or
(xxii) (a) a heavy chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 1,
CDR2 with the amino acid sequence of SEQ ID NO: 11 and
CDR3 with the amino acid sequence of SEQ ID NO: 17; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 31,
CDR2 with the amino acid sequence of SEQ ID NO: 39 and
CDR3 with the amino acid sequence of SEQ ID NO: 40.
In some embodiments of the invention, the isolated monoclonal antibody or antigen- binding fragment thereof includes a heavy chain variable domain that comprises an amino acid sequence that is selected from the group: SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 366, SEQ ID NO: 368, SEQ ID NO: 370 or SEQ ID NO: 372.
In some embodiments of the invention, the isolated monoclonal antibody or antigen- binding fragment thereof includes a light chain variable domain that comprises an amino acid sequence that is selected from the group: SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86 or SEQ ID NO: 87.
In some embodiments of the invention, the isolated monoclonal antibody or antigen- binding fragment thereof includes:
(a) a heavy chain variable domain that comprises an amino acid sequence selected from the group: SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 366, SEQ ID NO: 368, SEQ ID NO: 370 or SEQ ID NO: 372. and
(b) a light chain variable domain that comprises an amino acid sequence selected from the group: SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86 or SEQ ID NO: 87.
In some embodiments of the invention, the isolated monoclonal antibody or antigen- binding fragment thereof includes:
(i) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 44 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 74;
(ii) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 45 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 74;
(iii) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID
NO: 46 and (b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO:
75;
(iv) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 47 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 74;
(v) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 48 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 75;
(vi) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 49 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 74;
(vii) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 50 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 74;
(viii) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 51 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 74;
(ix) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 52 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 74;
(x) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 53 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 74;
(xi) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 54 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 76; (xii) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID
NO: 55 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 77;
(xiii) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 56 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 76;
(xiv) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 57 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 76;
(xv) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 58 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 76;
(xvi) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 59 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 76;
(xvii) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 53 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 78;
(xviii) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 60 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 75;
(xix) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 61 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 75;
(xx) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID
NO: 62 and (b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO:
75;
(xxi) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 63;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 75;
(xxii) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 64 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 75;
(xxiii) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 65 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 74;
(xxiv) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 66 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 75;
(xxv) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 67 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 74;
(xxvi) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 53 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 79;
(xxvii) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 68 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 75;
(xxviii) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 69 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO:
80; (xxix) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 70 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 81;
(xxx) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 71 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 75;
(xxxi) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 72 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 82;
(xxxii) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 73 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 74;
(xxxiii) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 53 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 83;
(xxxiv) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 53 and
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 84;
(xxxv) (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 53 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 85;
(xxxvi) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 53 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 86;
(xxxvii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 53;
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 87;
(xxxviii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 366; (b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74;
(xxxix) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 368;
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74;
(xxxx) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 370;
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74; or
(xxxxi) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 372;
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is a full-length IgG antibody.
In some embodiments of the invention, the isolated monoclonal antibody is a full-length IgG antibody that is of human IgGl, IgG2, IgG3 or IgG4 isotype.
In some embodiments of the invention, the isolated monoclonal antibody is a full-length IgG antibody that is of human IgG4 isotype.
In some embodiments of the invention, the isolated monoclonal antibody comprises the mutation S228P in the hinge region. The above mutation in the hinge region is numbered according to the EU numbering for amino acid chains of antibodies (Edelman, G.M., et al., Proc. Natl. Acad. Sci. USA 63 (1969), pp. 78-85; Kabat, E.A., et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD, (1991). The same mutation can be designated as S241P according to the Kabat numbering scheme for numbering amino acids of antibody chains, or as S229P according to sequential numbering from the beginning of the chain.
In some embodiments of the invention, the isolated monoclonal antibody includes a heavy chain that comprises an amino acid sequence that is selected from the group: SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 367, SEQ ID NO: 369, SEQ ID NO: 371 or , SEQ ID NO: 373. In some embodiments of the invention, the isolated monoclonal antibody includes a light chain that comprises an amino acid sequence that is selected from the group: SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130 or SEQ ID NO: 131.
In some embodiments of the invention, the isolated monoclonal antibody includes:
(i) (a) a heavy chain comprising an amino acid sequence that is selected from the group: SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 367, SEQ ID NO: 369, SEQ ID NO: 371 or , SEQ ID NO: 373, and
(b) a light chain comprising an amino acid sequence that is selected from the group: SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130 or SEQ ID NO: 131;
(ii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 88, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(iii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 89, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(iv) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 90, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 119;
(v) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 91, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(vi) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 92, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 119;
(vii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 93, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(viii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 94, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(ix) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 95, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118,
(x) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 96, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(xi) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(xii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 98, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 120,
(xiii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 99, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 121;
(xiv) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 100, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 120;
(xv) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 101, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 120;
(xvi) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 102, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 120,
(xvii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 103, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 120;
(xviii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 122;
(xix) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 104, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 119;
(xx) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 105, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 119;
(xxi) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 106, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 119;
(xxii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 107, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 119;
(xxiii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 108, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 119;
(xxiv) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 109, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(xxv) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 110, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 119;
(xxvi) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 111, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(xxvii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 123; (xxviii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 112, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 119;
(xxix) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 113, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 124;
(xxx) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 114, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 125;
(xxxi)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 115, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 119;
(xxxii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 116, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 126;
(xxxiii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 117, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(xxxiv) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 127;
(xxxv) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 128;
(xxxvi) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 129;
(xxxvii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 130;
(xxxviii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 131;
(xxxix) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 367, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(xxxx) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 369, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(xxxxi) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 371, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118; or
(xxxxii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 373, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 118.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 01.
Antibody 01 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 88; and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 118.
Antibody 01 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 44;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 74.
Antibody 01 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 13, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 01 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 132,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 143, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 164,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 01 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 174,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 185, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 02. Antibody 02 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 89; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118.
Antibody 02 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO:;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO:.
Antibody 02 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 13, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 02 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 133,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 143, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 164,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 02 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 175,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 185, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210. In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 03.
Antibody 03 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 90; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 119.
Antibody 03 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 46;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 75.
Antibody 03 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 14, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 32,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 35,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 41.
Antibody 03 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 134,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 144, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 162,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 165,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 171.
Antibody 03 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 176,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 186, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 204, (ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 207,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 211.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 04.
Antibody 04 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 91; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118.
Antibody 04 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 47;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 74.
Antibody 04 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 13, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 04 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 135,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 143, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 164,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 04 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 177,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 185, and (b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 05.
Antibody 05 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 92; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 119.
Antibody 05 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 48;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 75.
Antibody 05 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 15, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 32,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 35,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 41.
Antibody 05 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 145, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 162,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 165,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 171.
Antibody 05 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178, (ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 187, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 204,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 207,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 211.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 06.
Antibody 06 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 93; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118.
Antibody 06 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 49;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 74.
Antibody 06 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 13, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 06 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 143, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 164,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 06 includes: (a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 185, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 07.
Antibody 07 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 94; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118.
Antibody 07 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 50;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 74.
Antibody 07 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 16, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 07 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 146, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 164, (iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 07 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 188, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 08.
Antibody 08 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 95; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118.
Antibody 08 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 51;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 74.
Antibody 08 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 13, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 08 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 137,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 143, and
(b) a light chain variable domain comprising: (i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 164,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 08 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 179,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 185, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 09.
Antibody 09 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 96; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118.
Antibody 09 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 52;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 74.
Antibody 09 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 13, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 09 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 137,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142, (iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 143, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 164,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 09 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 179,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 185, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 10.
Antibody 10 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118.
Antibody 10 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO:
53;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO:
74.
Antibody 10 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 17, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40. Antibody 10 includes:
(a) a heavy chain variable domain comprising: (i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 147, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 164,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 10 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 189, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 11.
Antibody 11 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 98; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 120.
Antibody 11 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 54;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 76.
Antibody 11 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 2,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 12,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 18, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 33,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 36,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 42. Antibody 11 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 148, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 163,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 166,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 172.
Antibody 11 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 190, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 205,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 208,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 212.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 12.
Antibody 12 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 99; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 121.
Antibody 12 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 55;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 77.
Antibody 12 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 3,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 19, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 33, (ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 37,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 42.
Antibody 12 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 138,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 149, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 163,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 167,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 172.
Antibody 12 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 180,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 191, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 205,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 209,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 212.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 13.
Antibody 13 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 100; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 120.
Antibody 13 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 56;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 76.
Antibody 13 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 4,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 20, and (b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 33,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 36,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 42. Antibody 13 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 138,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 150, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 163,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 166,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 172.
Antibody 13 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 180,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 192, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 205,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 208,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 212.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 14.
Antibody 14 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 101; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 120.
Antibody 14 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 57;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 76.
Antibody 14 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 5, (ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 21, and (b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 33,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 36,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 42. Antibody 14 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 139,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 151, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 163,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 166,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 172.
Antibody 14 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 181,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 193, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 205,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 208,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 212.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 15.
Antibody 15 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 102; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 120.
Antibody 15 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 58;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO:
76.
Antibody 15 includes: (a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 22, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 33,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 36,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 42.
Antibody 15 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 152, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 163,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 166,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 172.
Antibody 15 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 194, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 205,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 208,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 212.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 16.
Antibody 16 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 103; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 120.
Antibody 16 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 59; (b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO:
76.
Antibody 16 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 6,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 23, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 33,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 36,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 42.
Antibody 16 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 140,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 153, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 163,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 166,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 172.
Antibody 16 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 182,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 195, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 205,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 208,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 212.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 17.
Antibody 17 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 122.
Antibody 17 includes: (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO:
53;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 78.
Antibody 17 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 17, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 38,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 17 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 147, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 168,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 17 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 189, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 18.
Antibody 18 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 104; and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 119.
Antibody 18 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 60;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 75.
Antibody 18 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 362,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 363, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 32,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 35,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 41.
Antibody 18 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 138,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 364, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 162,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 165,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 171.
Antibody 18 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 180,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 365, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 204,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 207,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 211.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 19. Antibody 19 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 105; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 119.
Antibody 19 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO:;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO:.
Antibody 19 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 7,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 24, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 32,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 35,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 41.
Antibody 19 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 141,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 154, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 162,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 165,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 171.
Antibody 19 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 183,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 196, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 204,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 207,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 211. In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 20.
Antibody 20 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 106; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 119.
Antibody 20 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 62;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 75.
Antibody 20 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 8,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 25, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 32,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 35,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 41.
Antibody 20 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 138,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 155, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 162,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 165,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 171.
Antibody 20 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 180,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 197, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 204, (ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 207,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 211.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 21.
Antibody 21 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 107; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 119.
Antibody 21 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 63;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 75.
Antibody 21 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 9,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 21, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 32,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 35,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 41.
Antibody 21 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 141,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 151, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 162,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 165,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 171.
Antibody 21 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 183,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 193, and (b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 204,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 207,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 211.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 22.
Antibody 22 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 108; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 119.
Antibody 22 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 64;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 75.
Antibody 22 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 8,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 26, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 32,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 35,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 41.
Antibody 22 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 138,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 156, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 162,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 165,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 171.
Antibody 22 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 180, (ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 198, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 204,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 207,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 211.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 23.
Antibody 23 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 109; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118.
Antibody 23 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 65;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 74.
Antibody 23 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 17, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 23 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 147, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 164,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 23 includes: (a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 189, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 24.
Antibody 24 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 110; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 119.
Antibody 24 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 66;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 75.
Antibody 24 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 8,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 27, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 32,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 35,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 41.
Antibody 24 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 138,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 157, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 162,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 165, (iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 171.
Antibody 24 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 180,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 199, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 204,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 207,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 211.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 25.
Antibody 25 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 111; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118.
Antibody 25 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 67;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 74.
Antibody 25 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 17, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 25 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 147, and
(b) a light chain variable domain comprising: (i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 164,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 25 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 189, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 26.
Antibody 26 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 123.
Antibody 26 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 53;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 79.
Antibody 26 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 17, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 26 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142, (iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 147, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 164,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 26 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 189, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 27.
Antibody 27 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 112; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 119.
Antibody 27 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO:
68;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO:
75.
Antibody 27 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 10,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 28, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 32,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 35,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 41. Antibody 27 includes:
(a) a heavy chain variable domain comprising: (i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 141,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 158, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 162,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 165,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 171.
Antibody 27 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 183,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 200, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 204,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 207,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 211.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 28.
Antibody 28 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 113; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 124.
Antibody 28 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO:
69;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO:
80.
Antibody 28 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 29, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40. Antibody 28 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 159, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 164,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 28 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 201, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 29.
Antibody 29 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 114; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 125.
Antibody 29 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 70;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 81.
Antibody 29 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 17, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31, (ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 29 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 147, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 164,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 29 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 189, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 30.
Antibody 30 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 115; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 119.
Antibody 30 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 71;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 75.
Antibody 30 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 8,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 30, and (b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 32,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 35,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 41. Antibody 30 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 138,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 160, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 162,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 165,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 171.
Antibody 30 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 180,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 202, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 204,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 207,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 211.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 31.
Antibody 31 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 116; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 126.
Antibody 31 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 72;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 82.
Antibody 31 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1, (ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 17, and (b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40. Antibody 31 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 147, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 164,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 31 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 189, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 32.
Antibody 32 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 117; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118.
Antibody 32 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 73;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO:
74.
Antibody 32 includes: (a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 17, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 32 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 147, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 164,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 32 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 189, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 33.
Antibody 33 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 127.
Antibody 33 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 53; (b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO:
83.
Antibody 33 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 17, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 43.
Antibody 33 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 147, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 164,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 173.
Antibody 33 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 189, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 213.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 34.
Antibody 34 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 128.
Antibody 34 includes: (a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO:
53;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 84.
Antibody 34 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 17, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 39,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 34 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 147, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 169,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 34 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 189, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 35.
Antibody 35 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97; and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 129.
Antibody 35 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 53;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 85.
Antibody 35 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 17, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 35 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 147, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 164,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 35 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 189, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210.
In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 36. Antibody 36 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 130.
Antibody 36 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO:;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO:.
Antibody 36 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 17, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 36 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 147, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 164,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 36 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 189, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210. In some embodiments of the invention, the isolated monoclonal antibody that specifically binds to IL-4Rα is antibody 37.
Antibody 37 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 131.
Antibody 37 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO: 53;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO: 87.
Antibody 37 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 17, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 37 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 136,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 142,
(iii) CDR3 (Chothia) with the amino acid sequence of SEQ ID NO: 147, and
(b) a light chain variable domain comprising:
(i) CDR1 (Chothia) with the amino acid sequence of SEQ ID NO: 161,
(ii) CDR2 (Chothia) with the amino acid sequence of SEQ ID NO: 164,
(iii) CDR3 (Chothia) with the amino acid sequence SEQ ID NO: 170.
Antibody 37 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 178,
(ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 184,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 189, and
(b) a light chain variable domain comprising:
(i) CDR1 (IMGT) with the amino acid sequence of SEQ ID NO: 203, (ii) CDR2 (IMGT) with the amino acid sequence of SEQ ID NO: 206,
(iii) CDR3 (IMGT) with the amino acid sequence of SEQ ID NO: 210.
Antibody 38 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 367; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118.
Antibody 38 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO:;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO:
Antibody 38 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 13, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 39 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 369; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118.
Antibody 39 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO:;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO:
Antibody 39 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 13, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34, (iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 40 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 371; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118.
Antibody 40 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO:;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO:
Antibody 40 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 13, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40.
Antibody 41 includes:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 373; and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118.
Antibody 41 includes:
(a) a heavy chain variable domain that comprises the amino acid sequence of SEQ ID NO:;
(b) a light chain variable domain that comprises the amino acid sequence of SEQ ID NO:
Antibody 41 includes:
(a) a heavy chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 1,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 11,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 17, and
(b) a light chain variable domain comprising:
(i) CDR1 (Kabat) with the amino acid sequence of SEQ ID NO: 31,
(ii) CDR2 (Kabat) with the amino acid sequence of SEQ ID NO: 34,
(iii) CDR3 (Kabat) with the amino acid sequence of SEQ ID NO: 40. Antibody fragments
In certain circumstances, it is advisable to use antibody fragments rather than whole antibodies. The small sizes of the fragments contributes to rapid clearance thereof and may contribute to better penetration into dense tumors.
Various techniques have been developed for the production of antibody fragments. Traditionally, these fragments were derived via proteolytic digestion of intact antibodies (see e.g. Morimoto et al, Journal of Biochemical and Biophysical Methods, 24, 1992, pp. 107-117 and Brennan et al, Science, 229, 1985, p. 81). However, these fragments can now be produced directly by recombinant host cells. Fab, Fv and ScFv antibody fragments can be expressed in and secreted from E. coh. thus allowing to facilitate the production of large amounts of these fragments. Antibody fragments may be isolated from the antibody phage libraries. According to another embodiment, Fab'-SH fragments can be directly isolated from E. coll and chemically coupled to form F(ab')2 fragments (Carter et al., Bio/Technology, 10, 1992, p. 163-167). According to another approach, F(ab')2 fragments can be isolated directly from a recombinant host cell culture. Fab- and F(ab')2-fragments with increased in vivo half-life retaining epitope binding receptor residues have been described in US 5869046. Other techniques for the production of antibody fragments will be apparent to those skilled in the art. In other embodiments of the invention, the selected antibody is a single-stranded Fv fragment (scFv) (see WO 93/16185; US 5571894 and US 5587458). Fv and scFv are the only species with intact binding sites that are devoid of constant regions; as a result, they are suitable for reduced nonspecific binding during in vivo use. scFv fusion proteins may be constructed to yield fusion of an effector protein at either N- or C-terminus of an scFv (see Antibody Engineering, ed. Borrebaeck above). The antibody fragment may also be a "linear antibody", e.g. as described in U.S. 5641870.
Nucleic acid molecule
In one aspect, the present invention relates to a nucleic acid that encodes any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα.
In any of said embodiments, the nucleic acid molecules may be isolated.
The terms "nucleic acid", "nucleic sequence", "nucleic acid sequence", "polynucleotide", "oligonucleotide", "polynucleotide sequence" and "nucleotide sequence", used interchangeably in the present description, mean a precise sequence of nucleotides, modified or not, determining a fragment or a region of a nucleic acid, containing unnatural nucleotides or not, and being either a double-strand DNA or RNA, a single-strand DNA or RNA, or transcription products of said DNAs.
It should also be included here that the present invention does not relate to nucleotide sequences in their natural chromosomal environment, i.e. in a natural state. The sequences of the present invention have been isolated and/or purified, i.e., they were sampled directly or indirectly, for example by copying, their environment having been at least partially modified. Thus, isolated nucleic acids obtained by recombinant genetics, by means, for example, of host cells, or obtained by chemical synthesis should also be mentioned here.
A reference to a nucleotide sequence encompasses the complement thereof unless otherwise specified. Thus, a reference to a nucleic acid having a particular sequence should be understood as one which encompasses the complementary strand thereof with the complementary sequence thereof.
An "isolated" nucleic acid molecule is one which is identified and separated from at least one nucleic acid molecule-impurity, which the former is typically bound to in the natural source of antibody nucleic acid. An isolated nucleic acid molecule is different from the form or set in which it is found under natural conditions. Thus, an isolated nucleic acid molecule is different from a nucleic acid molecule that exists in cells under natural conditions.
In one aspect, the present invention relates to a nucleic acid molecule comprising a nucleotide sequence encoding an amino acid sequence selected from SEQ ID NO: 1-213, 362-373. A nucleic acid molecule may also comprise any combination of said nucleotide sequences.
As would be appreciated by those skilled in the art, because of the redundancy of the genetic code, a variety of different DNA sequences can encode the amino acid sequence of the light chain or heavy chain of the antibody according to the invention or fragments thereof (VH, VL, CDR, etc.). It is well within the skill of a person trained in the art to create these alternative DNA sequences encoding the same amino acid sequences. Such variant DNA sequences are within the scope of the present invention.
In some embodiments of the invention, the isolated nucleic acid is DNA.
The nucleic acid molecule of the invention may be isolated from any source that produces the monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα. In certain embodiments of the invention, the nucleic acid molecule of the invention may be synthesized, rather than isolated.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 01, and includes the nucleotide sequence with SEQ ID NO: 214.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 01, and includes the nucleotide sequence with SEQ ID NO: 215. In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 01, and includes the nucleotide sequence with SEQ ID NO: 216.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 01, and includes the nucleotide sequence with SEQ ID NO: 217.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 02, and includes the nucleotide sequence with SEQ ID NO: 218.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 02, and includes the nucleotide sequence with SEQ ID NO: 219.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 02, and includes the nucleotide sequence with SEQ ID NO: 220.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 02, and includes the nucleotide sequence with SEQ ID NO: 221.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 03, and includes the nucleotide sequence with SEQ ID NO: 222.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 03, and includes the nucleotide sequence with SEQ ID NO: 223.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 03, and includes the nucleotide sequence with SEQ ID NO: 224.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 03, and includes the nucleotide sequence with SEQ ID NO: 225.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 04, and includes the nucleotide sequence with SEQ ID NO: 226. In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 04, and includes the nucleotide sequence with SEQ ID NO: 227.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 04, and includes the nucleotide sequence with SEQ ID NO: 228.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 04, and includes the nucleotide sequence with SEQ ID NO: 229.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 05, and includes the nucleotide sequence with SEQ ID NO: 230.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 05, and includes the nucleotide sequence with SEQ ID NO: 231.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 05, and includes the nucleotide sequence with SEQ ID NO: 232.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 05, and includes the nucleotide sequence with SEQ ID NO: 233.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 06, and includes the nucleotide sequence with SEQ ID NO: 234.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 06, and includes the nucleotide sequence with SEQ ID NO: 235.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 06, and includes the nucleotide sequence with SEQ ID NO: 236.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 06, and includes the nucleotide sequence with SEQ ID NO: 237. In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 07, and includes the nucleotide sequence with SEQ ID NO: 238.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 07, and includes the nucleotide sequence with SEQ ID NO: 239.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 07, and includes the nucleotide sequence with SEQ ID NO: 240.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 07, and includes the nucleotide sequence with SEQ ID NO: 241.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 08, and includes the nucleotide sequence with SEQ ID NO: 242.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 08, and includes the nucleotide sequence with SEQ ID NO: 243.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 08, and includes the nucleotide sequence with SEQ ID NO: 244.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 08, and includes the nucleotide sequence with SEQ ID NO: 245.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 09, and includes the nucleotide sequence with SEQ ID NO: 246.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 09, and includes the nucleotide sequence with SEQ ID NO: 247.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 09, and includes the nucleotide sequence with SEQ ID NO: 248. In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 09, and includes the nucleotide sequence with SEQ ID NO: 249.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 10, and includes the nucleotide sequence with SEQ ID NO: 250.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 10, and includes the nucleotide sequence with SEQ ID NO: 251.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 10, and includes the nucleotide sequence with SEQ ID NO: 252.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 10, and includes the nucleotide sequence with SEQ ID NO: 253.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 11, and includes the nucleotide sequence with SEQ ID NO: 254.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 11, and includes the nucleotide sequence with SEQ ID NO: 255.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 11, and includes the nucleotide sequence with SEQ ID NO: 256.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 11, and includes the nucleotide sequence with SEQ ID NO: 257.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 12, and includes the nucleotide sequence with SEQ ID NO: 258.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 12, and includes the nucleotide sequence with SEQ ID NO: 259. In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 12, and includes the nucleotide sequence with SEQ ID NO: 260.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 12, and includes the nucleotide sequence with SEQ ID NO: 261.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 13, and includes the nucleotide sequence with SEQ ID NO: 262.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 13, and includes the nucleotide sequence with SEQ ID NO: 263.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 13, and includes the nucleotide sequence with SEQ ID NO: 264.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 13, and includes the nucleotide sequence with SEQ ID NO: 265.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 14, and includes the nucleotide sequence with SEQ ID NO: 266.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 14, and includes the nucleotide sequence with SEQ ID NO: 267.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 14, and includes the nucleotide sequence with SEQ ID NO: 268.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 14, and includes the nucleotide sequence with SEQ ID NO: 269.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 15, and includes the nucleotide sequence with SEQ ID NO: 270. In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 15, and includes the nucleotide sequence with SEQ ID NO: 271.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 15, and includes the nucleotide sequence with SEQ ID NO: 272.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 15, and includes the nucleotide sequence with SEQ ID NO: 273.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 16, and includes the nucleotide sequence with SEQ ID NO: 274.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 16, and includes the nucleotide sequence with SEQ ID NO: 275.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 16, and includes the nucleotide sequence with SEQ ID NO: 276.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 16, and includes the nucleotide sequence with SEQ ID NO: 277.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 17, and includes the nucleotide sequence with SEQ ID NO: 278.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 17, and includes the nucleotide sequence with SEQ ID NO: 279.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 17, and includes the nucleotide sequence with SEQ ID NO: 280.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 17, and includes the nucleotide sequence with SEQ ID NO: 281. In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 18, and includes the nucleotide sequence with SEQ ID NO: 282.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 18, and includes the nucleotide sequence with SEQ ID NO: 283.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 18, and includes the nucleotide sequence with SEQ ID NO: 284.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 18, and includes the nucleotide sequence with SEQ ID NO: 285.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 19, and includes the nucleotide sequence with SEQ ID NO:286.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 19, and includes the nucleotide sequence with SEQ ID NO: 287.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 19, and includes the nucleotide sequence with SEQ ID NO: 288.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 19, and includes the nucleotide sequence with SEQ ID NO: 289.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 20, and includes the nucleotide sequence with SEQ ID NO: 290.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 20, and includes the nucleotide sequence with SEQ ID NO: 291.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 20, and includes the nucleotide sequence with SEQ ID NO: 292. In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 20, and includes the nucleotide sequence with SEQ ID NO: 293.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 21, and includes the nucleotide sequence with SEQ ID NO: 294.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 21, and includes the nucleotide sequence with SEQ ID NO: 295.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 21, and includes the nucleotide sequence with SEQ ID NO: 296.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 21, and includes the nucleotide sequence with SEQ ID NO: 297.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 22, and includes the nucleotide sequence with SEQ ID NO: 298.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 22, and includes the nucleotide sequence with SEQ ID NO: 299.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 22, and includes the nucleotide sequence with SEQ ID NO: 300.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 22, and includes the nucleotide sequence with SEQ ID NO: 301.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 23, and includes the nucleotide sequence with SEQ ID NO: 302.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 23, and includes the nucleotide sequence with SEQ ID NO: 303. In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 23, and includes the nucleotide sequence with SEQ ID NO: 304.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 23, and includes the nucleotide sequence with SEQ ID NO: 305.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 24, and includes the nucleotide sequence with SEQ ID NO: 306.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 24, and includes the nucleotide sequence with SEQ ID NO: 307.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 24, and includes the nucleotide sequence with SEQ ID NO: 308.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 24, and includes the nucleotide sequence with SEQ ID NO: 309.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 25, and includes the nucleotide sequence with SEQ ID NO: 310.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 25, and includes the nucleotide sequence with SEQ ID NO: 311.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 25, and includes the nucleotide sequence with SEQ ID NO: 312.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 25, and includes the nucleotide sequence with SEQ ID NO: 313.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 26, and includes the nucleotide sequence with SEQ ID NO: 314. In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 26, and includes the nucleotide sequence with SEQ ID NO: 315.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 26, and includes the nucleotide sequence with SEQ ID NO: 316.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 26, and includes the nucleotide sequence with SEQ ID NO: 317.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 27, and includes the nucleotide sequence with SEQ ID NO: 318.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 27, and includes the nucleotide sequence with SEQ ID NO: 319.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 27, and includes the nucleotide sequence with SEQ ID NO: 320.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 27, and includes the nucleotide sequence with SEQ ID NO: 321.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 28, and includes the nucleotide sequence with SEQ ID NO: 322.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 28, and includes the nucleotide sequence with SEQ ID NO: 323.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 28, and includes the nucleotide sequence with SEQ ID NO: 324.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 28, and includes the nucleotide sequence with SEQ ID NO: 325. In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 29, and includes the nucleotide sequence with SEQ ID NO: 326.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 29, and includes the nucleotide sequence with SEQ ID NO: 327.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 29, and includes the nucleotide sequence with SEQ ID NO: 328.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 29, and includes the nucleotide sequence with SEQ ID NO: 329.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 30, and includes the nucleotide sequence with SEQ ID NO: 330.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 30, and includes the nucleotide sequence with SEQ ID NO: 331.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 30, and includes the nucleotide sequence with SEQ ID NO: 332.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 30, and includes the nucleotide sequence with SEQ ID NO: 333.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 31, and includes the nucleotide sequence with SEQ ID NO: 334.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 31, and includes the nucleotide sequence with SEQ ID NO: 335.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 31, and includes the nucleotide sequence with SEQ ID NO: 336. In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 31, and includes the nucleotide sequence with SEQ ID NO: 337.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 32, and includes the nucleotide sequence with SEQ ID NO: 338.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 32, and includes the nucleotide sequence with SEQ ID NO: 339.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 32, and includes the nucleotide sequence with SEQ ID NO: 340.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 32, and includes the nucleotide sequence with SEQ ID NO: 341.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 33, and includes the nucleotide sequence with SEQ ID NO: 342.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 33, and includes the nucleotide sequence with SEQ ID NO: 343.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 33, and includes the nucleotide sequence with SEQ ID NO: 344.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 33, and includes the nucleotide sequence with SEQ ID NO: 345.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 34, and includes the nucleotide sequence with SEQ ID NO: 346.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 34, and includes the nucleotide sequence with SEQ ID NO: 347. In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 34, and includes the nucleotide sequence with SEQ ID NO: 348.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 34, and includes the nucleotide sequence with SEQ ID NO: 349.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 35, and includes the nucleotide sequence with SEQ ID NO: 350.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 35, and includes the nucleotide sequence with SEQ ID NO: 351.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 35, and includes the nucleotide sequence with SEQ ID NO: 352.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 35, and includes the nucleotide sequence with SEQ ID NO: 353.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 36, and includes the nucleotide sequence with SEQ ID NO: 354.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 36, and includes the nucleotide sequence with SEQ ID NO: 355.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 36, and includes the nucleotide sequence with SEQ ID NO: 356.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 36, and includes the nucleotide sequence with SEQ ID NO: 357.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 37, and includes the nucleotide sequence with SEQ ID NO: 358. In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 37, and includes the nucleotide sequence with SEQ ID NO: 359.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 37, and includes the nucleotide sequence with SEQ ID NO: 360.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 37, and includes the nucleotide sequence with SEQ ID NO: 361.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 38, and includes the nucleotide sequence with SEQ ID NO: 374.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 38, and includes the nucleotide sequence with SEQ ID NO: 215.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 38, and includes the nucleotide sequence with SEQ ID NO: 375.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 38, and includes the nucleotide sequence with SEQ ID NO: 217.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 39, and includes the nucleotide sequence with SEQ ID NO: 376.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 39, and includes the nucleotide sequence with SEQ ID NO: 215.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 39, and includes the nucleotide sequence with SEQ ID NO: 377.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 39, and includes the nucleotide sequence with SEQ ID NO: 217. In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 40, and includes the nucleotide sequence with SEQ ID NO: 378.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 40, and includes the nucleotide sequence with SEQ ID NO: 215.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 40, and includes the nucleotide sequence with SEQ ID NO: 379.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 40, and includes the nucleotide sequence with SEQ ID NO: 217.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain variable domain of antibody 41, and includes the nucleotide sequence with SEQ ID NO: 380.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain variable domain of antibody 41, and includes the nucleotide sequence with SEQ ID NO: 215.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the heavy chain of antibody 41, and includes the nucleotide sequence with SEQ ID NO: 381.
In some embodiments of the invention, the nucleic acid is a nucleic acid that encodes the amino acid sequence of the light chain of antibody 41, and includes the nucleotide sequence with SEQ ID NO: 217.
The nucleic acid molecules may be used to express the monoclonal antibody or antigen- binding fragment thereof that specifically binds to IL-4Rα.
Vector
In one aspect, the present invention relates to an expression vector comprising the above isolated nucleic acid. The present invention relates to a vector suitable for the expression of any of nucleotide sequences described herein.
The term "vector" as used herein means a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. In some embodiments of the invention, the vector is a plasmid, i.e. a circular double stranded piece of DNA into which additional DNA segments may be ligated. In some embodiments of the invention, the vector is a viral vector, wherein additional DNA segments may be ligated into the viral genome. In some embodiments of the invention, vectors are capable of autonomous replication in a host cell into which they are introduced (e.g. bacterial vectors having a bacterial site of replication origin and episomal mammalian vectors). In further embodiments of the invention, vectors (e.g. non-episomal mammalian vectors) may be integrated into the genome of a host cell upon introduction into a host cell, and thereby are replicated along with the host gene. Moreover, certain vectors are capable of directing the expression of genes to which they are operably linked. Such vectors are referred to herein as "recombinant expression vectors" (or simply, "expression vectors").
The present invention relates to vectors comprising nucleic acid molecules that encode any of the amino acid sequences of the monoclonal antibody that specifically binds to IL-4Rα or portions thereof (e.g. heavy chain and/or light chain binding domain sequences), as described herein. The invention further relates to vectors comprising nucleic acid molecules encoding antibodies or fragments of these antibodies.
Expression vectors include plasmids, retroviruses, adenoviruses, adeno-associated viruses (AAVs), plant viruses, such as cauliflower mosaic virus, tobacco mosaic virus, cosmids, YACs, EBV derived episomes, and the like. DNA molecules may be ligated into a vector such that transcriptional and translational control sequences within the vector serve their intended function of regulating the transcription and translation of the DNA. An expression vector and expression control sequences may be chosen to be compatible with the expression host cell used. DNA molecules partially or fully encoding the sequences of first and second binding domains (for example, heavy and light chain sequences where a binding domain comprises a heavy and light chain sequence) may be introduced into individual vectors. In one embodiment, any combination of the above DNA molecules is introduced into the same expression vector. DNA molecules may be introduced into an expression vector by standard methods (e.g. ligation of complementary restriction sites on a gene fragment of antibody and vector, or blunt end ligation if no restriction sites are present).
In some embodiments of the invention, a suitable vector is one that includes restriction sites such that any VH or VL sequence can easily be inserted and expressed, as described above. Polyadenylation and transcription termination may occur at a native chromosomal site downstream of coding regions. A recombinant expression vector can also encode a signal peptide that facilitates secretion of an antibody chain from a host cell. An antibody chain gene may be cloned into a vector such that the signal peptide is linked in-frame to the amino terminus of an immunoglobulin chain. The signal peptide may be an immunoglobulin signal peptide or a heterologous signal peptide (i.e. a signal peptide from a non-immunoglobulin protein).
In some embodiments of the invention, in addition to antibody chain genes, the recombinant expression of the vectors according to the invention can carry regulatory sequences that control the expression of antibody chain genes in a host cell. It will be understood by those skilled in the art that the design of an expression vector, including the selection of regulatory sequences, may depend on such factors as the choice of a host cell to be transformed, the level of expression of a desired protein, and so forth. Preferred control sequences for an expression host cell in mammals include viral elements that ensure high levels of protein expression in mammalian cells, such as promoters and/or enhancers derived from a retroviral LTR, cytomegalovirus (CMV) (such as a CMV promoter/enhancer), simian virus 40 (SV40) (such as a SV40 promoter/enhancer), adenovirus, (e.g. the major late promoter adenovirus (AdMLP)), polyomavirus and strong mammalian promoters such as native immunoglobulin promoter or actin promoter. Methods for expressing polypeptides in bacterial cells or fungal cells, e.g. yeast cells, are also well known in the art.
In some embodiments of the invention, in addition to antibody chain genes and regulatory sequences, the recombinant expression vectors of the invention may carry additional sequences, such as sequences that regulate replication of a vector in host cells (e.g. origins of replication) and selectable marker genes. The selectable marker gene facilitates the selection of host cells into which a vector has been introduced.
In some embodiments of the invention, the vector may include an expression control sequence. The term "expression control sequence" as used in the present description refers to polynucleotide sequences that are necessary to effect the expression and processing of coding sequences to which they are ligated. The expression control sequences include appropriate transcription initiation, termination, promoter and enhancer sequences; efficient RNA processing signals such as splicing and polyadenylation signals; sequences that stabilize cytoplasmic mRNA; sequences that enhance translation efficiency (i.e., Kozak consensus sequence); sequences that enhance protein stability; and when desired, sequences that enhance protein secretion. The nature of such control sequences differs depending upon the host organism; in prokaryotes, such control sequences generally include a promoter, a ribosome binding site, and transcription termination sequences; in eukaryotes, such control sequences typically include promoters and transcription termination sequences. The term "control sequences" includes at least all components, the presence of which is essential for expression and processing, and may further include additional components, the presence of which is advantageous, for example, leader sequences and fusion partner sequences.
Host cell
In one aspect, the present invention relates to a method for producing a host cell to produce any above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα, and comprises transformation of the cell with the above vector. In one aspect, the present invention relates to a host cell for producing any above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα, which comprises any of the above nucleic acids.
The term "recombinant host cell" (or simply "host cell") as used herein refers to a cell into which a recombinant expression vector has been introduced. The present invention relates to host cells, which may include, for example, the above-described vector according to the invention. The present invention further relates to host cells that comprise, for example, a nucleotide sequence encoding a heavy chain or antigen-binding portions thereof, a nucleotide sequence encoding a light chain or antigen-binding portions thereof, or both, of the binding domain of the binding molecule according to the invention. It should be understood that "recombinant host cell" and "host cell" refer not only to a particular subject cell but to the progeny of such cell as well. Since modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to a parental cell; however, such cells are still included within the scope of the term "host cell" as used herein.
The nucleic acid molecules encoding the monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα according to the invention and vectors comprising these nucleic acid molecules may be used for transfection of a suitable mammalian or cell thereof, plant or cell thereof, bacterial or yeast host cell. Transformation may be carried out by any known technique of introducing polynucleotides into a host cell. Methods for introduction of heterologous polynucleotides into mammalian cells are well known in the art and include dextran-mediated transfection, cationic polymer-nucleic acid complex transfection, calcium phosphate precipitation, polybrene-mediated transfection, protoplast fusion, encapsulation of the polynucleotides in liposomes, and direct microinjection of DNA into nuclei. In addition, the nucleic acid molecules may be introduced into mammalian cells by viral vectors.
Mammalian cell lines used as hosts for transformation are well known in the art and include a plurality of immortalized cell lines available. These include, e.g., Chinese hamster ovary (CHO) cells, NSO cells, SP2 cells, HEK-293T cells, FreeStyle 293 cells (Invitrogen), NH4-3T3 cells, HeLa cells, baby hamster kidney (BHK) cells, African green monkey kidney cells (COS), human hepatocellular carcinoma cells (e.g., Hep G2), A549 cells, and a number of other cell lines. Cell lines are selected by determining which cell lines have high expression levels and provide for necessary characteristics of the protein being produced. Other cell lines that may be used are insect cell lines, such as Sf9 or Sf21 cells. When the recombinant expression vectors encoding the monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα are introduced into mammalian host cells, the antibodies or fragments thereof are produced by culturing the host cells for a period of time sufficient to allow for expression of the antibodies or fragments thereof in host cells or, more preferably, secretion of the antibodies or fragments thereof into the culture medium in which the host cells are grown. The monoclonal antibody or antigen- binding fragment thereof that specifically binds to IL-4Rα may be isolated from the culture medium using conventional protein purification techniques. Plant host cells include e.g. Nicotiana, Arabidopsis, duckweed, corn, wheat, potato, etc. Bacterial host cells include Escherichia and Streptomyces species. Yeast host cells include Schizosaccharomyces pombe, Saccharomyces cerevisiae and Pichia pastoris.
Furthermore, the level of production of the monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα from a production cell line may be enhanced using a number of known techniques. For example, the glutamine synthetase gene expression system (the GS system) is a common approach for enhancing expression under certain conditions.
It is likely that the monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα from various cell lines will have a different glycosylation profile as compared to each other. However, the monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα encoded by nucleic acid molecules described herein, or comprising amino acid sequences provided herein are part of the present invention, regardless of the glycosylation of the binding molecules, and, in general, regardless of the presence or absence of post-translational modifications.
The above host cell does not refer to a host cell produced using human embryos.
The above host cell does not refer to a host cell produced by modifying the genetic integrity of human germline cells.
Method of producing the antibody
In one aspect, the present invention relates to a method for producing an antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα, comprising culturing the above host cell in a growth medium under conditions sufficient to produce said antibody or fragment thereof, if necessary, followed by isolation and purification of the resulting antibody or fragment thereof.
The present invention relates to methods for producing the monoclonal antibody or antigen- binding fragment thereof that specifically binds to IL-4Rα according to the present invention. One embodiment of the invention relates to a method for producing the monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα, as defined herein, which comprises producing a recombinant host cell capable of expressing the monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα, culturing said host cell under conditions suitable for expression/production of the monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα, and isolating the resulting monoclonal antibody or fragment thereof that specifically binds to IL-4Rα. The monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα produced by such expression in such recombinant host cells is referred to herein as "recombinant monoclonal antibody that specifically binds to IL-4Rα" or "antigen-binding fragment of the recombinant monoclonal antibody that specifically binds to IL-4Rα".
Pharmaceutical compositions
Another aspect of the invention is a pharmaceutical composition comprising, as an active ingredient (or as the only active ingredient), the monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα according to the present invention.
In one aspect, the present invention relates to a pharmaceutical composition used for treating a disease or disorder mediated by IL-4Rα, which comprises any above antibody or antigen- binding fragment thereof in a therapeutically effective amount in combination with one or more pharmaceutically acceptable excipients.
"Pharmaceutical composition" refers to a composition comprising an antibody of the present invention and at least one of components selected from the group comprising pharmaceutically acceptable and pharmacologically compatible fillers, solvents, diluents, carriers, auxiliary, distributing and sensing agents, delivery agents, such as preservatives, stabilizers, filler, disintegrators, moisteners, emulsifiers, suspending agents, thickeners, sweeteners, flavouring agents, aromatizing agents, antibacterial agents, fungicides, lubricants, and prolonged delivery controllers, the choice and suitable proportions of which depend on the type and way of administration and dosage. Examples of suspending agents are ethoxylated isostearyl alcohol, polyoxyethene, sorbitol and sorbitol ether, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacant and mixtures thereof as well. Protection against action of microorganisms can be provided by various antibacterial and antifungal agents, such as, for example, parabens, chlorobutanole, sorbic acid, and similar compounds. The composition may also contain isotonic agents, such as, for example, sugars, polyols, sodium chloride, and the like. Prolonged action of the composition may be achieved by agents slowing down absorption of active ingredient, for example, aluminum monostearate and gelatine. Examples of suitable carriers, solvents, diluents and delivery agents are water, ethanol, polyalcohols and mixtures thereof, plant oils (such as olive oil) and organic esters (such as ethyl oleate) for injections. Examples of fillers are lactose, milk sugar, sodium citrate, calcium carbonate, calcium phosphate, and the like. Examples of disintegrators and distributors are starch, alginic acid and salts thereof, silicates. Examples of lubricants are magnesium stearate, sodium lauryl sulfate, talc, and polyethylene glycol of high molecular weight as well. The pharmaceutical composition for peroral, sublingual, transdermal, intraocular, intramuscular, intravenous, subcutaneous, local or rectal administration of active ingredient, alone or in combination with another active compound, may be administered to human and animals in a standard administration form, in a mixture with traditional pharmaceutical carriers. Suitable standard administration forms include peroral forms such as tablets, gelatin capsules, pills, powders, granules, chewing gums and peroral solutions or suspensions; sublingual and transbuccal administration forms; aerosols; implants; local, transdermal, subcutaneous, intramuscular, intravenous, intranasal or intraocular administration forms and rectal administration forms.
The term "excipient" is used herein to describe any ingredient other than the antibody according to the present invention. These are substances of inorganic or organic nature which are used in the pharmaceutical production/manufacturing in order to give drug products the necessary physicochemical properties.
In some embodiments, the compositions are intended to improve, prevent, or treat diseases or disorders that may be mediated by IL-4Rα.
The term "disease or disorder mediated by IL-4Rα" refers to any disease or disorder that is either directly, or indirectly associated with IL-4Rα, including etiology, development, progression, persistence or pathology of a disease or disorder.
"Treat", "treatment" and "therapy" refer to a method of alleviating or abrogating a biological disorder and/or at least one of attendant symptoms thereof. Further, references herein to "treatment" include references to curative, palliative and prophylactic treatment.
In one aspect, the subject of treatment, or patient, is a mammal, preferably a human subject. Said subject may be either male or female, of any age.
The term "disorder" means any condition that would benefit from treatment according to the present invention. The definition of the term includes chronic and acute disorders or diseases including those pathological conditions that predispose the mammal to the disorder in question.
"Therapeutically effective amount" refers to that amount of the therapeutic agent being administered during treatment which will relieve to some extent one or more of the symptoms of the disease being treated.
The pharmaceutical compositions of the present invention and methods of preparation thereof will be undoubtedly apparent to those skilled in the art. The pharmaceutical compositions should preferably be manufactured in compliance with the GMP (Good Manufacturing Practice) requirements. The composition may comprise a buffer composition, tonicity agents, stabilizers and solubilizers. Prolonged action of composition may be achieved by agents slowing down absorption of an active pharmaceutical ingredient, for example, aluminum monostearate and gelatine. Examples of suitable carriers, solvents, diluents and delivery agents include water, ethanol, polyalcohols and mixtures thereof, oils, and organic esters for injections.
Any method for administering peptides, proteins or antibodies which is accepted in the art may be suitably employed for the monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα according to the present invention.
The term "pharmaceutically acceptable" refers to one or more compatible liquid or solid components that are suitable for administration in a mammal, preferably in a human.
The terms "buffer", "buffer composition", "buffering agent" refers to a solution, which is capable of resisting changes in pH by the action of its acid-base conjugate components, and which allows the product of antibody that specifically binds to IL-4Rα to resist changes in pH. Generally, the pharmaceutical composition preferably has a pH in the range from 4.0 to 8.0. Examples of buffers used include, but are not limited to, acetate, phosphate, citrate, histidine, succinate, etc. buffer solutions.
The terms "tonic agent", "osmolyte" or "osmotic agent", as used herein, refer to an excipient that can increase the osmotic pressure of a liquid antibody formulation. "Isotonic" drug is a drug that has an osmotic pressure equivalent to that of human blood. Isotonic drugs typically have an osmotic pressure from about 250 to 350 mOsm/kg. Isotonic agents used include, but are not limited to, polyols, saccharides and sucrose, amino acids, metal salts, for example, sodium chloride, etc.
"Stabilizer" refers to an excipient or a mixture of two or more excipients that provide the physical and/or chemical stability of the active agent. Stabilizers may be amino acids, for example, but not limited to, arginine, histidine, glycine, lysine, glutamine, proline; surfactants, for example, but not limited to, polysorbate 20 (trade name: Tween 20), polysorbate 80 (trade name: Tween 80), polyethylene-polypropylene glycol and copolymers thereof (trade names: Pol oxamer, Pluronic, sodium dodecyl sulfate (SDS); antioxidants, for example, but not limited to, methionine, acetylcysteine, ascorbic acid, monothioglycerol, sulfurous acid salts, etc.; chelating agents, for example, but not limited to, ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTP A), sodium citrate, etc.
The pharmaceutical composition according to the invention is a stable composition.
The pharmaceutical composition is "stable" if the active agent retains physical stability and/or chemical stability and/or biological activity thereof during the specified shelflife at storage temperature, for example, of 2-8 °C. Preferably, the active agent retains both physical and chemical stability, as well as biological activity. Storage period is adjusted based on the results of stability test in accelerated or natural aging conditions. A pharmaceutical composition according to the invention may be manufactured, packaged, or widely sold in the form of a single unit dose or a plurality of single unit doses in the form of a ready formulation. The term "single unit dose" as used herein refers to discrete quantity of a pharmaceutical composition containing a predetermined quantity of an active ingredient. The quantity of an active ingredient typically equals the dose of the active ingredient to be administered in a subject, or a convenient portion of such dose, for example, half or a third of such dose.
The pharmaceutical compositions according to the present invention are typically suitable for parenteral administration as sterile formulations intended for administration in a human body through the breach in skin or mucosal barriers, bypassing the gastrointestinal tract by virtue of injection, infusion and implantation. In particular, it is contemplated that parenteral administration includes, inter alia, subcutaneous, intraperitoneal, intramuscular, intravenous, intraarterial, intrathecal, intraventricular, intraurethral, intracranial, intrasynovial, transdermal injection or infusion; and kidney dialytic infusion techniques. Intra-tumor delivery, for example, intra-tumor injection, may also be employed. Regional perfusion is also contemplated. Preferred embodiments include intravenous and subcutaneous routes. The most preferred method of administration of the pharmaceutical composition according to the present invention is subcutaneous administration. Any method for administering peptides or proteins which is accepted in the art may be suitably employed for the antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα according to the present invention.
Injectable formulations may be prepared, packaged, or sold, without limitation, in unit dosage form, such as in ampoules, vials, plastic containers, pre-filled syringes, autoinjection devices. Formulations for parenteral administration include, inter alia, suspensions, solutions, emulsions in oily or aqueous bases, pastes, and the like.
In another embodiment, the invention provides a composition for parenteral administration comprising a pharmaceutical composition which is provided in dry (i.e. powder or granular) form for reconstitution in a suitable solvent (e.g. sterile pyrogen-free water) prior to administration. Such medicinal formulation may be prepared by, for example, lyophilization, i.e. a process, which is known in the art as freeze drying, and which involves freezing a product followed by removal of solvent from frozen material.
The antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα according to the present invention may also be administered intranasally or by inhalation, either alone, as a mixture with a suitable pharmaceutically acceptable vehicle from an inhaler, such as a pressurised aerosol container, pump, spray, atomiser, or nebuliser, where a suitable propellant is used or not used, or as nasal drops, or spray. Medicinal formulations for parenteral administration may be formulated to be immediate or modified release. Modified release medicinal formulations include delayed, sustained, pulsed, controlled, targeted and programmed release.
In one aspect, the present invention relates to a pharmaceutical composition for treating a disease or disorder mediated by IL-4Rα, the pharmaceutical combination comprising any above antibody or antigen-binding fragment thereof and at least one other therapeutically active compound.
In one aspect, the present invention relates to a pharmaceutical composition for treating a disease or disorder mediated by IL-4Rα, the pharmaceutical combination comprising any above antibody or antigen-binding fragment thereof and at least one other therapeutically active compound, which is an antibody, a small molecule, a hormone therapy agent or a combination thereof.
In some embodiments of the pharmaceutical composition, the disease or disorder mediated by IL-4Rα is selected from the group: atopic dermatitis, eczema, allergic dermatitis, atopic keratoconjunctivitis, bronchial asthma, chronic obstructive pulmonary disease, allergic bronchopulmonary aspergillosis, chronic rhinosinusitis with nasal polyps, chronic rhinosinusitis, fungal rhinosinusitis, nasal polyps, sleep apnea, allergic rhinitis, Churg-Strauss syndrome, itching associated with chronic liver diseases, hypertrophic scar, alopecia areata, urticaria, allergic dermatitis, focal scleroderma, pemphigus, Netherton syndrome, ichthyosis, eosinophilic gastritis, eosinophilic esophagitis, prurigo nodularis, colitis, filariasis, leishmaniasis, schistosomiasis, arterial hypertension, interstitial lung diseases, metastatic non-small cell lung cancer, metastases of epithelial tumors, malignant neoplasm of the prostate gland, malignant neoplasm of the brain, stomach cancer, colorectal cancer, malignant neoplasm of the colon, malignant neoplasm of the breast, pancreatic cancer, malignant neoplasm of the extrahepatic bile ducts, intrahepatic bile duct cancer, malignant neoplasm of the gallbladder, Hodgkin lymphoma, medulloblastoma, hypereosinophilic syndrome, allergic reactions, acute respiratory distress syndrome.
Therapeutic use of monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα
In one aspect, the antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is used in the treatment of disorders mediated by IL-4Rα activity.
In one aspect, the subject of treatment, or patient, is a mammal, preferably a human subject. Said subject may be either male or female, of any age.
In one aspect, the present invention relates to a method for treatment of a disease or disorder mediated by IL-4Rα, which comprises administering in a subject in need of such treatment any above antibody or antigen-binding fragment thereof or the above pharmaceutical composition in a therapeutically effective amount.
In one aspect, the present invention relates to a method for treatment of a disease or disorder mediated by IL-4Rα, which comprises administering in a subject in need of such treatment any above antibody or antigen-binding fragment thereof and at least one other therapeutically active compound in a therapeutically effective amount.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is selected from the group: atopic dermatitis, eczema, allergic dermatitis, atopic keratoconjunctivitis, bronchial asthma, chronic obstructive pulmonary disease, allergic bronchopulmonary aspergillosis, chronic rhinosinusitis with nasal polyps, chronic rhinosinusitis, fungal rhinosinusitis, nasal polyps, sleep apnea, allergic rhinitis, Churg-Strauss syndrome, itching associated with chronic liver diseases, hypertrophic scar, alopecia areata, urticaria, allergic dermatitis, focal scleroderma, pemphigus, Netherton syndrome, ichthyosis, eosinophilic gastritis, eosinophilic esophagitis, prurigo nodularis, colitis, filariasis, leishmaniasis, schistosomiasis, arterial hypertension, interstitial lung diseases, metastatic non-small cell lung cancer, metastases of epithelial tumors, malignant neoplasm of the prostate gland, malignant neoplasm of the brain, stomach cancer, colorectal cancer, malignant neoplasm of the colon, malignant neoplasm of the breast, pancreatic cancer, malignant neoplasm of the extrahepatic bile ducts, intrahepatic bile duct cancer, malignant neoplasm of the gallbladder, Hodgkin lymphoma, medulloblastoma, hypereosinophilic syndrome, allergic reactions, acute respiratory distress syndrome.
In some embodiments of the method for treatment, the other therapeutically active compound is a small molecule, hormone therapy agent, or any combination thereof.
In one aspect, the present invention relates to the use of the above antibody or antigen- binding fragment thereof or the above pharmaceutical composition for treating in a subject in need of such treatment a disease or disorder mediated by IL-4Rα.
In one aspect, the present invention relates to the use of the above antibody or antigen- binding fragment thereof and at least one other therapeutically active compound for treating a disease or disorder mediated by IL-4Rα.
In some embodiments of the use, the disease or disorder mediated by IL-4Rα is selected from the group: atopic dermatitis, eczema, allergic dermatitis, atopic keratoconjunctivitis, bronchial asthma, chronic obstructive pulmonary disease, allergic bronchopulmonary aspergillosis, chronic rhinosinusitis with nasal polyps, chronic rhinosinusitis, fungal rhinosinusitis, nasal polyps, sleep apnea, allergic rhinitis, Churg-Strauss syndrome, itching associated with chronic liver diseases, hypertrophic scar, alopecia areata, urticaria, allergic dermatitis, focal scleroderma, pemphigus, Netherton syndrome, ichthyosis, eosinophilic gastritis, eosinophilic esophagitis, prurigo nodularis, colitis, filariasis, leishmaniasis, schistosomiasis, arterial hypertension, interstitial lung diseases, metastatic non-small cell lung cancer, metastases of epithelial tumors, malignant neoplasm of the prostate gland, malignant neoplasm of the brain, stomach cancer, colorectal cancer, malignant neoplasm of the colon, malignant neoplasm of the breast, pancreatic cancer, malignant neoplasm of the extrahepatic bile ducts, intrahepatic bile duct cancer, malignant neoplasm of the gallbladder, Hodgkin lymphoma, medulloblastoma, hypereosinophilic syndrome, allergic reactions, acute respiratory distress syndrome.
In some embodiments of the use, the other therapeutically active compound is a small molecule, hormone therapy agent, or any combination thereof.
For the treatment of the above diseases, a monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα according to the present invention may be administered in combination with another therapeutic agent, such as an immunosuppressor, an anti-inflammatory medicinal product, a systemic hormonal medicinal product, an antineoplastic and immunomodulatory medicinal product, or others, using a multidrug regimen.
The uses or methods used herein relating to the antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα with one or more other therapeutic agents are contemplated to mean, refer to and include the following:
1) simultaneous administration of such combination of the antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα and therapeutic agent to a patient in need of treatment, when such components are formulated together into a single dosage form which releases said components at substantially the same time to said patient,
2) simultaneous administration of such combination of the antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα and therapeutic agent to a patient in need of treatment, when such components are formulated apart from each other into separate dosage forms which are taken at substantially the same time by said patient, whereupon said components are released at substantially the same time to said patient,
3) sequential administration of such combination of the antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα and therapeutic agent to a patient in need of treatment, when such components are formulated apart from each other into separate dosage forms which are taken at consecutive times by said patient with a significant time interval between each administration, whereupon said components are released at substantially different times to said patient; and
4) sequential administration of such combination of the antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα and therapeutic agent to a patient in need of treatment, when such components are formulated together into a single dosage form which releases said components in a controlled manner, whereupon they are concurrently, consecutively, or jointly released at the same and/or different times to said patient, where each portion may be administered by either the same or different routes.
The antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα may be administered without further therapeutic treatment, i.e. as an independent therapy.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is atopic dermatitis (eczema, nummular eczema).
In some embodiments of the method for treatment of atopic dermatitis (eczema, nummular eczema), any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment as monotherapy or in combination with topical agents (moisturizers and emollients, topical glucocorticoids (GCs), topical calcineurin inhibitors, topical antibiotics and antiseptics), systemic GCs, immunosuppressants (cyclosporine, azathioprine, methotrexate, mycophenolate mofetil, interferon gamma), oral calcineurin inhibitors, oral antihistamines, monoclonal antibodies against TSLP or any combination thereof.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is bronchial asthma (aspirin-induced asthma, aspirin triad).
In some embodiments of the method for treatment of bronchial asthma (aspirin-induced asthma, aspirin triad), any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment as monotherapy or in combination with inhaled and/or oral GCs, beta2-agonists, muscarinic receptor antagonists, leukotriene receptor antagonists, methylxanthines, monoclonal antibodies against immunoglobulin E, monoclonal antibodies against interleukin 5, monoclonal antibodies against interleukin 5 receptor, monoclonal antibodies against TSLP or any combination thereof.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is chronic obstructive pulmonary disease.
In some embodiments of the method for treatment of chronic obstructive pulmonary disease, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment as monotherapy or in combination with beta2-agonists, methylxanthines, GCs, phosphodiesterase-4 inhibitors, muscarinic receptor antagonists or any combination thereof.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is allergic bronchopulmonary aspergillosis.
In some embodiments of the method for treatment of Allergic bronchopulmonary aspergillosis, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment in combination with antifungal drugs, systemic GCs or any combination thereof.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is chronic rhinosinusitis with nasal polyps, chronic rhinosinusitis, fungal rhinosinusitis, nasal polyps or allergic rhinitis.
In some embodiments of the method for treatment of chronic rhinosinusitis with nasal polyps, chronic rhinosinusitis, fungal rhinosinusitis, nasal polyps or allergic rhinitis, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment as monotherapy or in combination with antifungal agents (for fungal rhinosinusitis), intranasal GCs, monoclonal antibodies against interleukin 5, monoclonal antibodies against interleukin 5 receptor, monoclonal antibodies against TSLP or any combination thereof.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is Churg-Strauss syndrome.
In some embodiments of the method for treatment of Churg-Strauss syndrome, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment as monotherapy or in combination with GCs, immunosuppressants (cyclophosphamide, azathioprine, methotrexate, mycophenolate mofetil), immunoglobulins, intranasal GCs, monoclonal antibodies against interleukin 5, monoclonal antibodies against interleukin 5 receptor, monoclonal antibodies against CD20 (for example, rituximab) or any combination thereof.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is itching associated with chronic liver diseases.
In some embodiments of the method for treatment of itching associated with chronic liver diseases, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment as monotherapy or in combination with therapy for underlying liver disease.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is hypertrophic scar.
In some embodiments of the method for treatment of hypertrophic scar, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment as monotherapy or in combination with GCs.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is alopecia areata or focal scleroderma. In some embodiments of the method for treatment of alopecia areata or focal scleroderma, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment as monotherapy or in combination with GCs, minoxidil, anthralin, immunosuppressants (cyclophosphamide, azathioprine, methotrexate, my cophenolate mofetil), TNF-alpha inhibitors or any combination thereof.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is urticaria.
In some embodiments of the method for treatment of urticaria, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment as monotherapy or in combination with antihistamines, leukotriene receptor antagonists, immunosuppressants (cyclosporine), monoclonal antibodies against immunoglobulin E or any combination thereof.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is pemphigus.
In some embodiments of the method for treatment of pemphigus, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment in combination with GCs, immunosuppressants (cyclosporine, azathioprine, methotrexate, mycophenolate mofetil, chlorambucil), intravenous immunoglobulins, monoclonal antibodies against immunoglobulin E, monoclonal antibodies against CD20 or any combination thereof.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is Netherton syndrome.
In some embodiments of the method for treatment of Netherton syndrome, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment as monotherapy or in combination with symptomatic therapy (where various groups of medicinal products depending on clinical manifestations can be used).
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is ichthyosis.
In some embodiments of the method for treatment of ichthyosis, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment as monotherapy or in combination with retinoids.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is atopic keratoconjunctivitis.
In some embodiments of the method for treatment of atopic keratoconjunctivitis, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment as monotherapy or in combination with GCs, calcineurin inhibitors or any combination thereof.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is prurigo nodularis.
In some embodiments of the method for treatment of prurigo nodularis, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment as monotherapy or in combination with GCs, calcineurin inhibitors, immunosuppressants (cyclosporine, methotrexate), thalidomide, calcipotriol, capsaicin, antiepileptic medications and antidepressants or any combination thereof.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is colitis.
In some embodiments of the method for treatment of colitis, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment as monotherapy or in combination with GCs, TNF-alpha inhibitors, vedolizumab, tofacitinib, ustekinumab, methotrexate, sulfasalazine or any combination thereof.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is filariasis, leishmaniasis or schistosomiasis.
In some embodiments of the method for treatment of filariasis, leishmaniasis, schistosomiasis, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment in combination with antiparasitic agents.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is arterial hypertension.
In some embodiments of the method for treatment of arterial hypertension, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment in combination with antihypertensive medications, diuretics or any combination thereof.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is interstitial lung disease.
In some embodiments of the method for treatment of interstitial lung diseases, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment in combination with GCs, antifibrotic medications or any combination thereof.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is metastatic non-small cell lung cancer or other oncological diseases. In some embodiments of the method for treatment of metastatic non-small cell lung cancer or other oncological diseases, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment in combination with immunotherapy, targeted therapy, chemotherapy or any combination thereof.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is hypereosinophilic syndrome.
In some embodiments of the method for treatment of hypereosinophilic syndrome, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment in combination with GCs, monoclonal antibodies against interleukin 5, monoclonal antibodies against interleukin 5 receptor, monoclonal antibodies against CD52 receptor, or any combination thereof.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is allergic reactions.
In some embodiments of the method for treatment of allergic reactions, any of the above antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is administered to the subject in need of such treatment as monotherapy or in combination with GCs, antihistamines or any combination thereof.
In some embodiments of the method for treatment, the disease or disorder mediated by IL- 4Rα is acute respiratory distress syndrome (for example, SARS-cov-2-induced acute respiratory distress syndrome).
In some embodiments of the method for treatment of acute respiratory distress syndrome (for example, SARS-cov-2-induced acute respiratory distress syndrome), any of the above antibody or antigen-binding fragment that specifically binds to IL-4Rα is administered to the subject in need of such treatment in combination with etiotropic therapy, anti -CO VID convalescent plasma, human immunoglobulin against COVID-19, anti-inflammatory therapy (GCs, monoclinal antibodies), antithrombotic therapy or any combination thereof.
Doses and routes of administration
The monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL- 4Rα according to the invention will be administered in an amount that is effective in treatment of the condition in question, i.e. in doses and during the periods of time required to achieve the desired result. A therapeutically effective amount may vary according to factors such as the particular condition being treated, the age, sex and weight of the patient, and whether the monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα is being administered as a stand-alone treatment or in combination with one or more additional drugs or treatments. Dosage regimens may be adjusted to provide the optimum desired response. For example, a single bolus may be administered, several divided doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is especially advantageous to formulate parenteral compositions in a unit dosage form for ease of administration and uniformity of dosage. A unit dosage form as used herein refers to physically discrete units suited as unitary dosages for patients/subjects to be treated; each unit contains a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the desired pharmaceutical carrier. Specification for the unit dosage forms of the invention is typically dictated by and directly dependent on (a) the unique characteristics of a therapeutic agent and particular therapeutic or prophylactic effect to be achieved, and (b) the limitations inherent in the art of compounding such an active compound for the treatment of sensitivity in subjects.
Thus, a skilled artisan would appreciate, based upon the disclosure provided herein, that the doses and dosage regimen are adjusted in accordance with methods well-known in the therapeutic arts. That is, the maximum tolerable dose can be readily established, and the effective amount providing a detectable therapeutic effect to a patient may also be determined, as can the temporal requirements for administering each agent to provide a detectable therapeutic effect to a patient. Thus, while certain doses and administration regimens are exemplified herein, these examples in no way limit the doses and administration regimens that may be provided to a patient in practicing the embodiments of the invention.
It is to be noted that dosage values may vary with the type and severity of the condition to be alleviated, and may include single or multiple doses. Furthermore, it is to be understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the judgment of a medical professional administering or supervising the administration of the compositions, and that dosage ranges set forth in the present description are exemplary only and are not intended to limit the scope or practice of the claimed compositions. Further, the dosage regimen with the compositions according to the present invention may be based on a variety of factors, including the type of disease, the age, weight, sex, medical condition of the patient, the severity of the condition, the route of administration, and the particular monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα employed. Thus, the dosage regimen may vary widely, but can be determined routinely using standard methods. For example, doses may be adjusted based on pharmacokinetic and pharmacodynamic parameters, which may include clinical effects such as toxic effects or laboratory values. Thus, the present invention encompasses intra-patient dose-escalation as determined by one skilled in the art. Methods for determining appropriate dosage and regimens are well-known in the art and would be understood by a skilled artisan once provided the ideas disclosed herein.
Examples of suitable administration methods have been provided above.
It is believed that a suitable dose of a monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα according to the invention will be in the range of 0.1-200 mg/kg, preferably 0.1-100 mg/kg, including about 0.5-50 mg/kg, for example about 1-20 mg/kg. The monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα may be administered, e.g. in a dose of at least 0.25 mg/kg, such as at least 0.5 mg/kg, including at least 1 mg/kg, e.g. at least 1.5 mg/kg, such as at least 2 mg/kg, e.g. at least 3 mg/kg, including at least 4 mg/kg, e.g. at least 5 mg/kg; and for example up to a maximum of 50 mg/kg, including up to a maximum of 30 mg/kg, e.g. up to a maximum of 20 mg/kg, including up to a maximum of 15 mg/kg. The administration will typically be repeated in appropriate time intervals, such as once a week, once every two weeks, once every three weeks or once every four weeks, and for as long as deemed appropriate by a responsible physician, who may, in some cases, increase or reduce the dose if necessary.
Diagnostic use of antibody that specifically binds to IL-4Ra
The monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL- 4Rα according to the invention are also used in diagnostic purposes (e.g. in vitro, ex vivo). For example, the present monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα according to the present invention may be used for detecting or measuring the level of IL-4Rα in samples obtained from a patient (e.g. tissue sample or a sample of body fluid, such as an inflammatory exudate, blood, serum, intestinal fluid, saliva or urine). Suitable methods for detection and measurement include immunoassays, such as flow cytometry, enzyme-linked immunosorbent assay (ELISA), chemiluminescent assay, radioimmunoassay, and immunohi stol ogy .
Examples
The following examples are provided for better understanding of the invention. These examples are for purposes of illustration only and are not to be construed as limiting the scope of the invention in any manner.
All publications, patents, and patent applications cited in this specification are incorporated herein by reference. Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended embodiments. Materials and general methods
General information regarding the nucleotide sequences of human immunoglobulin light and heavy chains is given in: Kabat, E.A., et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD (1991). Amino acids of antibody chains are numbered according to EU numbering (Edelman, G.M., et al., Proc. Natl. Acad. Sci. USA 63 (1969) 78-85; Kabat, E.A., et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD, (1991).
Recombinant DNA techniques
Standard methods were used to manipulate DNA as described in Sambrook, J. et al, Molecular cloning: A laboratory manual; Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1989. The molecular biological reagents were used according to the manufacturer protocols.
Gene synthesis
Desired gene segments were prepared from oligonucleotides made by chemical synthesis. The gene segments of 300-1400 bp long, which were flanked by singular restriction sites, were assembled by annealing and ligation of oligonucleotides including PCR amplification and subsequently cloned via the restriction sites. The DNA sequences of the subcloned gene fragments were confirmed by DNA sequencing.
DNA sequence determination
DNA sequences were determined by Sanger sequencing.
DNA and protein sequence analysis and sequence data management
The Unipro's UGENE suite version 1.29 and SnapGene Viewer were used for sequence creation, mapping, analysis, annotation and illustration.
Expression vectors
For the expression of the antibodies described in the application materials, variants of expression plasmids intended for expression of antibodies in prokaryotic cells (E.coli), transient expression in eukaryotic cells (e.g., in CHO cells) were applied. Beside the antibody expression cassette the vectors contained: an origin of replication which allows replication of said plasmid in E. coll. genes which confer resistance in E. coli to various antibiotics (e.g. to ampicillin, kanamycin).
The fusion genes comprising the described antibody chains as described below were generated by PCR and/or gene synthesis and assembled with known recombinant methods and techniques by connection of the according nucleic acid segments, e.g. using unique restriction sites in the corresponding vectors. The subcloned nucleic acid sequences were verified by DNA sequencing. For transient transfections, larger quantities of the plasmids were prepared by plasmid preparation from transformed E. coll cultures.
Example 1. Production of recombinant antigens in suspension culture of mammalian cells.
Sequences of human extracellular domains of the IL-4Rα receptor (interleukin-4 receptor subunit alpha) (SEQ https://www.uniprot.org/uniprot/P24394 (27-227 aa)) were cloned into the pEE plasmids to generate Fclama-, His-Flag-tagged protein in mammalian cells using Sall/Notl restriction sites. The required quantities of plasmids were cultured in E.Coli cells and purified using the Qiagen kit.
The sequence of extracellular domain of Cynomolgus monkey IL-4Rα (SEQ https://www.uniprot.org/uniprot/A0A2K5V580 (24-232 aa)) was cloned into the pEE plasmid to generate Fc-tagged protein in mammalian cells using Sall/Notl restriction sites. The required quantities of plasmids were cultured in E.coli cells and purified using Qiagen kit.
Example 2. Production of recombinant antigens in suspension culture of mammalian cells
Antigens were produced in established cell line obtained from Chinese hamster ovary (CHO-T) cells. Suspension culture was conducted in flasks on orbital incubator shaker using serum-free media supplemented with 8 mM L-glutamine and 1 g/1 pluronic 68. For transient expression, cells at 1.9-2.2>< 106 cells/ml were transfected using linear polyethyleneimine. DNA/PEI ratio was 1:6/1 :7. 9 days following transfection, culture liquid was separated from cells by filtration through a 0.5/0.22 pm deep-bed filter.
The IL-4Rα-Fclama, cIL-4Rα-Fclama (cynomolgus) antigen was isolated from the culture fluid and purified using a column with a Protein A affinity chromatography sorbent. The cleared culture liquid was passed through a 20 ml column equilibrated with phosphate buffered saline (PBS, pH 7.4). The column was then washed with 5 column volumes of PBS to remove non- specifically binding components. Bound antigen was eluted using 0.1 M glycine buffer (pH 2.5.). The principal protein elution peak was collected and brought to neutral pH with 1 M Tris buffer (pH 7.5). Protein was then dialyzed into PBS (pH 7.4), ImM DTT was added, the mixture was filtered through 0.22 pm Millex GP, transferred into tubes and stored at -70°C.
0.5 M Imidazole and 0.1 M NiC12 were added to the cleared culture liquid containing IL- 4Rα-6HisFlag antigens to a concentration of 0.02 M Imidazole and 0.001 M NiC12. The culture liquid was then passed through a 10 ml column equilibrated with phosphate buffered saline supplemented with 10 mM imidazole (PBS, pH 7.4). The column was then washed with 5 column volumes of PBS supplemented with 10 mM imidazole to remove non-specifically binding components. Bound antigen was eluted with PBS supplemented with 500 mM imidazole (pH 7.4). The protein was then dialyzed into PBS. Thereafter, the protein was applied onto a gel filtration column pre-equilibrated with PBS. The protein was eluted with PBS, filtered through 0.22 pm Millex GP, transferred to tubes and stored at -70 °C.
Antigen electrophoresis was performed in a denaturing 10% polyacrylamide gel under non- reducing conditions without mercaptoethanol and under reducing conditions with mercaptoethanol.
Example 3. Cloning of genes of antibody variable domains into expression plasmid
The developed gene sequences of variable domains of antibodies according to the invention were cloned into the expression plasmid pLL (Fig. 1) according to a standard protocol using the restriction enzyme ligase method.
Next, the expression vectors containing antibody fragments were transformed into the E. coli BL21-Gold strain.
Example 4. Generation and primary analysis of Fab fragments of antibodies 1-37 specifically binding human IL-4Rα
Fabs were generated according to the standard technique: E. coli BL21-Gold bacterial cells were transformed with expression vectors containing Fab genes, and the subsequent addition of an inducer triggered transcription of lac operon, thus, when culturing the resulting transformants, causing expression of Fabs which were exported into periplasmic space. ELISA was then performed to verify Fab binding to substrate-immobilized IL-4Rα-Fclama antigen at a concentration of 0.2 pg/ml on (high binding) plates in 0.1 M NaHCOs (pH 9.0) (the antigen was immobilized overnight at 4 °C). All further steps were carried out at room temperature according to the standard ELISA protocol. Washing was performed after each step, with 300 pl/well lx PBST in three replications. Non-specific binding sites on the plate were blocked with 1% fat-free milk in lx PBST; analyte (60 pl/well of E. coli supernatants) was added following washing. Immune complexes were detected using peroxidase-conjugated goat anti -Fab antibodies (1 :7500). Substrate-chromogenic mixture was stained by adding 50 pl of substrate solution (H202-0.02% and TMB in CTLCOONa (pH 5.5)) for 15 minutes. 25 pl of 1% H2SO4 was used to stop the reaction. Color signal was measured at 450 nm using the suitable Tecan Sunrise plate reader. Antibody binding level was proportional to the signal produced.
Thus, all the test Fab fragments of antibodies 1-37 showed specific binding to IL-4Rα- Fclama.
Example 5. Analysis of Fab fragments to human IL-4Rα antigen on Forte Bio
Experiments involving the investigation of the affinity of purified Fab fragments were carried out on the Forte Bio instrument using FAB2G biosensors (ForteBio) that enable the immobilization of Fab of antibodies onto a CH1 domain region. The analysis was carried out at 30 °C using a kinetic buffer solution (4.3 mM Na2HPO4; 136.9 mM NaCl, 1.5 mM KH2PO4; 2.7 mM KC1; the volume fraction of the added Tween 20 was 0.1%; the mass fraction of the added BSA was 0.1%; pH 7.4), this buffer was used at all stages of the analysis. The Fab samples were immobilized onto sensors for 600 seconds, the concentration was 20 pg/ml. The baseline was recorded for 300 seconds. Following the baseline recording, the sensors with immobilized Fab were immersed into wells containing the analyte solution (IL-4Rα-His), where the antigen-Fab complex was associated for 300 seconds. The concentration of IL-4Rα-His antigen was 400 nM, the reference signal of the kinetic buffer solution free of IL-4Rα-His was recorded for each Fab. The complex dissociation in buffer solution was then detected for 600 seconds. To check the nonspecific interaction between the analyte and the sensors (negative control), we used sensors free of Fab fragments of antibodies. At the loading step, the negative control sensors were immersed into Fab-free sodium acetate buffer, and all other steps were similar to those used for the sensors loaded with Fab samples).
Binding curves were analyzed using the Octet Data Analysis (Version 9.0) software and using the 1 : 1 interaction model (Table 2).
Table 2 - Dissociation rate constants for antibodies (01-37) when interacting with human IL-4Rα- His antigen; Response is signal level reached at the end of the association step; negative control is verification of nonspecific interaction between the analyte (IL-4Rα -His) and sensors not loaded with Fab fragments
Figure imgf000108_0001
Figure imgf000109_0001
IL-4Rα-His antigen (Table 2).
Example 6. Analysis of non-specific binding of generated Fabs to other antigens
We employed ELISA to analyze nonspecific binding of test Fab fragments to other antigens. IL5R-Fclama, INFB2a, CD3-FLAG-EPEA in 0.1 M NaHCO3 (pH 9.0) were used as immobilization antigens (the antigen was immobilized overnight at 4 °C). IL-4Rα-Fclama, IL- 4Rα-His were used as a specific binding control (the antigen was immobilized overnight at 4°C). All further steps were carried out at room temperature according to the standard ELISA protocol. Washing was performed after each step, with 300 pl/well lx PBST in three replications. Non- specific binding sites on the plate were blocked with 1% fat-free milk in lx PBST; analyte (60 pl/well of E. coll supernatants) was added following washing. Immune complexes were detected using peroxidase-conjugated goat anti-Fab antibodies (1 :7500). Substrate-chromogenic mixture was stained by adding 50 pl of substrate solution (H202-0.02% and TMB in CHsCOONa (pH 5.5)) for 15 minutes. 25 pl of 1% H2SO4 was used to stop the reaction. Color signal was measured at 450 nm using the suitable Tecan Sunrise plate reader. Antibody binding level was proportional to the signal produced. Thus, all test Fab fragments of antibodies 1-37 showed specific binding to IL-4Rα-Fclama, IL-4Rα-His and did not show non-specific interaction with IL5R-Fclama, INFp2a, CD3-FLAG- EPEA.
Example 7. Production of genetic constructs to synthesize full-length antibodies
Cloning was performed by the standard technique. We generated PCR products comprising the genes of heavy and light chain variable domains of antibodies, with primers comprising restriction sites. The heavy chain variable domain (VH) was cloned into the vector pEE-HC IgG4 (Fig. 2) using the Sall/Nhel restriction sites. The light chain variable domain (VL) was cloned into the vector pEE-CK (Fig. 3) via Sall/BsiWl restriction sites. The resulting gene constructs were transferred for transient production of proteins in CHO-T cell line.
Example 8. Production of full-length antibodies
Full-length antibodies were produced in established cell line obtained from Chinese hamster ovary cells (CHO-T line) by transient transfection in two replicates. Suspension culture was conducted in orbital shake bioreactors using serum-free media supplemented with 8 mM L- glutamine and 1 g/1 Pluronic 68. For transient expression, cells at 2-2.2* 106 cells/ml were transfected using linear polyethyleneimine. DNA/PEI ratio was 1 :7. On day 10 of culturing, the cell suspension was centrifuged under 2000 g for 15 min and filtered through 0.22 pm filter.
Antibodies were purified by affinity chromatography columns using a robotic station. The column was equilibrated with a buffer containing 50 mM NaPB, 150 mM NaCl (pH 7.5), the filtered culture liquid with antibodies was applied, the column was then washed with 8 volumes of a buffer containing 50 mM NaPB, 150 mM NaCl and 4 volumes of 50 mM NaPB (pH 7.5). Protein was eluted with 6 column volumes of a solution of 50 mM NaPB, 100 mM NaCl (pH 3). The resulting samples were neutralized with 25 μl of IM phosphate buffer (pH 8).
Polyacrylamide gel electrophoresis and size-exclusion HPLC were used to control the purity of the antibodies. Electrophoresis was performed in 7.5% polyacrylamide gel under denaturing non-reducing conditions. The protein purity was determined by the intensity of band staining at a protein load of 10 pg per lane. Electrophoregrams are shown in Fig. 4, Fig. 5, Fig.6, Fig.7, Fig.8. Size-exclusion HPLC was performed on a column in a mobile phase of 0.05M NaH2PO4, 0.3 M NaCl pH=7.0.
Example 9. Determination of affinity of full-length antibodies to human IL-4Rα on Forte Bio
Experimental study of antibody binding affinity to the human IL-4Rα-His antigen was performed on the Forte Bio instrument. Antibodies at a concentration of 20 pg/ml were immobilized onto the surface of Protein A biosensors (ForteBio). The analysis was carried out at 30 °C using a kinetic buffer solution (4.3 mM Na2HPO4; 136.9 mM NaCl, 1.5 mM KH2PO4; 2.7 mM KC1; the volume fraction of the added Tween 20 was 0.1%; the mass fraction of the added BSA was 0.1%; pH 7.4). After setting the baseline in the kinetic buffer solution, the sensors with immobilized antibodies were dipped into wells containing the analyte (IL-4Rα-His) solution, where the association of the complex took place for 300 seconds. For each test antibody, we recorded three sensorgrams for the solutions of IL-4Rα-His having the following concentrations: 96.2 nM, 48.1 nM, 24.0 nM, and a reference signal (reference sensorgram) of a kinetic buffer solution free of IL-4Rα-His. The complex dissociation in buffer solution was then detected for 600 seconds. To check the nonspecific interaction between the analyte and the sensors (negative control), we used sensors free of antibodies. At the loading step, the negative control sensors were immersed into antibody-free sodium acetate buffer, and all other steps were similar to those used for the sensors loaded with antibodies).
Binding curves, after subtracting a reference signal, were analyzed using the Octet Data Analysis (Version 9.0) software and using 1 : 1 Global interaction model.
Table 3 - Kinetic constants for antibodies 01 - 37 when interacting with the human IL-4Rα-His antigen; Response is signal level reached at the end of the association step, negative control is verification of the nonspecific interaction between the analyte (IL-4Rα -His) and sensors not loaded with antibodies.
Figure imgf000111_0001
Figure imgf000112_0001
Figure imgf000113_0001
Figure imgf000114_0001
Thus, all test anti-IL-4Rα antibodies specifically bind to the human IL-4Rα antigen (Table
3).
Example 10. Examination of interactions between full-length antibodies and cynomolgus cIL-4Rα on Forte Bio.
This experiment aimed at confirming the interaction between the antibodies and the cynomolgus cIL-4Rα-Fclama antigen (macaca fascicularis, IL-4Rα with Fc fragment and 6His tag at the C-terminus). The interaction between antibodies and the cynomolgus cIL-4Rα-Fclama antigen was verified on the Forte Bio instrument using AR2G biosensors (ForteBio). The experiment consisted of the following steps: activating sensors, loading protein onto sensors, quenching unreacted activated groups, recording baselines, recording analyte association, recording dissociation. The sensors were activated in an aqueous solution comprising 20 mM EDC and 10 mM sNHS for 300 s. The antibodies were loaded onto the surface of biosensors in a sodium- acetate buffer solution with pH 5.0 for 300 s. The concentration of protein to be loaded was 20 pg/ml. Unreacted active centers on the sensor surfaces were quenched in IM aqueous solution of ethanolamine with pH 8.5 for 300 s. To verify the nonspecific interaction between the analyte and the sensors (negative control), we used an antibody-free sensor (at the loading step, the sensor was immersed in sodium acetate buffer solution with pH 5.0; all other steps were similar to those used for the sensors loaded with antibodies). The baseline and all subsequent steps of the experiment were carried out in a kinetic buffer solution (4.3 mM Na2HPO4; 136.9 mM NaCl, 1.5 mM KH2PO4; 2.7 mM KC1; the volume fraction of added Tween 20 was 0.1%; the mass fraction of added BSA was 0.1%; pH 7.4). At the association step (step duration was 300 s), sensors loaded with protein were dipped into wells containing a solution of analyte (IL-4Rα-Fclama) prepared in a kinetic buffer. The analyte concentration was 197.2 nM. At the dissociation step, the sensors were immersed into wells with a kinetic buffer, where the baseline was recorded. The measurements were carried out at 30 °C.
Binding curves were processed using the Octet Data Analysis (Version 9.0) software and using the 1 : 1 interaction model. The results of processing are shown in Table 4 (the value of kdis <1.0E-07 (1/s) and the corresponding thereto value of KD <1.0E-12 (M) means that, within the framework of the applied measurement and processing technique, the given sample shows exceeded limit of sensitivity of the dissociation rate constant for the model curve describing the sensorgram). In view of the use of a bivalent analyte (the antigen is present in the solution in the dimeric form due to the presence of an Fc fragment), the resulting values of the kinetic constants are evaluative and can only be used to compare samples. All anti-IL-4Rα antibodies showed a signal of binding to the cynomolgus cIL-4Rα-Fclama antigen at the association step and a slow decline in the signal at the dissociation step, thereby confirming the interaction with this antigen.
Table 4 - Results of processing of sensorgrams of binding of antibodies 01 - 37 to the cIL-4Rα- Fclama antigen. The value of the constant Kdis <1.0E-07 (1/s) and the corresponding thereto value of KD <1.OE-12 (M) means that, within the framework of the applied measurement and processing technique, the given sample shows exceeded limit of sensitivity of the dissociation rate constant for the model curve describing the sensorgram. Response is the signal level reached at the end of the association step; negative control is verification of the nonspecific interaction between the analyte and sensors not loaded with antibodies. The given values of the constants KD, Kon and Kdis can only be used to compare antibodies, in view of the use of a bivalent analyte.
Figure imgf000115_0001
Figure imgf000116_0001
Thus, all test anti-IL-4Rα antibodies specifically bind to the cynomolgus cIL-4Rα-Fclama antigen (Table 4).
Example 11. Inhibition of biological effect of hIL-4 and hIL-13 in vitro
The analysis is designed to evaluate the ability of antibodies to inhibit the hIL-4Rα- dependent cell cascade in vitro using the transgenic HEK-Blue IL4/IL13 cell line carrying STAT6 and a reporter construct (SEAP) located under the STAT6-inducible promoter.
The cells were plated into 96-well plates (5*104 cells/well), incubated for 4 hours, a pre- prepared antibody titer (serial dilution method, from 0 to 25 pg/ml) and a solution of hIL-4 (1 ng/ml) or hIL-13 (5 ng/ml) were introduced. The cells were incubated for 24 hours under standard conditions. The cellular response was evaluated by determining the activity of alkaline phosphatase. The results are shown in Table 5.
Table 5 - Inhibition of biological effect of hIL-4 and hIL-13 in vitro.
Figure imgf000117_0001
Figure imgf000118_0001
* not measured due to small protein amounts
Accordingly, all test anti-IL-4Rα antibodies inhibit hIL-4- and hIL- 13 -dependent signaling in vitro.
Example 12. Inhibition of hIL-4-dependent proliferation on TF-1 cell line.
The analysis has been designed to assess the ability of antibodies to inhibit TF-1 cell line proliferation mediated by the interaction between IL-4Rα Type 2 expressed on the membrane surface of TF-1 cells and hIL-4.
The cells were plated into 96-well plates (3*104 cells/well), a pre-prepared antibody titer (serial dilution method, from 0 to 100 pg/ml) and a solution of hIL-4 (100 ng/ml) were introduced. The cells were incubated for 3 days under standard conditions. The cellular response was evaluated using the Alamar blue viability dye according to the manufacturer's instructions. The results are shown in Table 6.
Table 6 - Inhibition of hIL-4-dependent proliferation on TF-1 cell line
Figure imgf000118_0002
Figure imgf000119_0001
* not measured due to small protein amounts
Accordingly, all test anti-IL-4Rα antibodies inhibit TF-1 cell line proliferation mediated by interaction between IL-4Rα Type 2 expressed on the membrane surface of the line cells

Claims

Claims
1. An isolated monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα (interleukin-4 receptor subunit alpha), comprising
(a) a heavy chain variable domain comprising:
(i) CDR1 with the amino acid sequence X1YWX2H, wherein
X1= T, Y, S, A or H;
X2 = M, F, L, I or V;
(ii) CDR2 with the amino acid sequence X3IRPAGSSTYYTDSVKG, wherein X3 = T or S; and
(iii) CDR3 with the amino acid sequence GGX4SX5X6WX7X8X9X10X11, wherein X4=Y, R or N;
X5= S, A or R;
X6 = T, Q, E, R or S;
X7= Y, H, G, T or V;
X8=R, V, K or S;
X9=Y, M, A, G, L or Q;
X10=D, S, E, G, Q, H or N;
X11=P, Y or S; and
(b) a light chain variable domain comprising:
(i) CDR1 with the amino acid sequence X12GSX13SNIGX14X15X16VG, wherein
X12 = S or T;
X13 = T, R or S;
X14 = S, G or D;
X15 = N or Y;
X16 = V, T or Y;
(ii) CDR2 with the amino acid sequence X17X18X19X20X21X22S, wherein
X17 = R, I or S;
X18 = D or N;
X19 = N, S or T;
X20= Q, E, N or R; X21= R or H;
X22= P or A; and
(iii) CDR3 with amino acid sequence X23X24WDDX25LX26X27VV, wherein
X23=S, A or T;
X24= T, A or S’
X25 = S or G;
X26 = N or S;
X27 = G or 0.
2. The isolated monoclonal antibody or antigen -binding fragment thereof according to claim 1, wherein the heavy chain variable domain comprises CDR1 with an amino acid sequence selected from the group: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10 or SEQ ID NO: 362.
3. The isolated monoclonal antibody or antigen -binding fragment thereof according to claim 1, wherein the heavy chain variable domain comprises CDR2 with an amino acid sequence selected from the group: SEQ ID NO: 11 or SEQ ID NO: 12.
4. The isolated monoclonal antibody or antigen -binding fragment thereof according to claim 1, wherein the heavy chain variable domain comprises CDR3 with an amino acid sequence selected from the group: SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30 or SEQ ID NO: 363.
5. The isolated monoclonal antibody or antigen -binding fragment thereof according to claim 1, wherein the light chain variable domain comprises CDR1 with an amino acid sequence selected from the group: SEQ ID NO: 31, SEQ ID NO: 32 or SEQ ID NO: 33.
6. The isolated monoclonal antibody or antigen -binding fragment thereof according to claim 1, wherein the light chain variable domain comprises CDR2 with an amino acid sequence selected from the group: SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38 or SEQ ID NO: 39.
7. The isolated monoclonal antibody or antigen -binding fragment thereof according to claim 1, wherein the light chain variable domain comprises CDR3 with an amino acid sequence selected from the group: SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42 or SEQ ID NO: 43.
8. The isolated monoclonal antibody or antigen -binding fragment thereof according to claim 1, comprising:
(a) a heavy chain variable domain comprising:
CDR1 with an amino acid sequence selected from the group: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10 or SEQ ID NO: 362;
CDR2 with an amino acid sequence selected from the group: SEQ ID NO: 11 or SEQ ID NO: 12; and
CDR3 with an amino acid sequence selected from the group: SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30 or SEQ ID NO: 363; and
(b) a light chain variable domain comprising:
CDR1 with an amino acid sequence selected from the group: SEQ ID NO: 31, SEQ ID NO: 32 or SEQ ID NO: 33;
CDR2 with an amino acid sequence selected from the group: SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38 or SEQ ID NO: 39; and
CDR3 with an amino acid sequence selected from the group: SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42 or SEQ ID NO: 43.
9. The isolated monoclonal antibody or antigen -binding fragment thereof according to claim 8, comprising:
(i) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 1, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 13; and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 31, CDR2 with the amino acid sequence of SEQ ID NO: 34 and CDR3 with the amino acid sequence of SEQ ID NO: 40;
(ii) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 1, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 14; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 32, CDR2 with the amino acid sequence of SEQ ID NO: 35 and CDR3 with the amino acid sequence of SEQ ID NO: 41;
(iii) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 1, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 15; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 32, CDR2 with the amino acid sequence of SEQ ID NO: 35 and CDR3 with the amino acid sequence of SEQ ID NO: 41;
(iv) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 1, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 16; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 31, CDR2 with the amino acid sequence of SEQ ID NO: 34 and CDR3 with the amino acid sequence of SEQ ID NO: 40;
(v) (a) a heavy chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 1, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 17; and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 31, CDR2 with the amino acid sequence of SEQ ID NO: 34 and CDR3 with the amino acid sequence of SEQ ID NO: 40;
(vi) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 2, CDR2 with the amino acid sequence of SEQ ID NO: 12 and CDR3 with the amino acid sequence of SEQ ID NO: 18; and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 33, CDR2 with the amino acid sequence of SEQ ID NO: 36 and CDR3 with the amino acid sequence of SEQ ID NO: 42;
(vii) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 3, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 19; and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 33, CDR2 with the amino acid sequence of SEQ ID NO: 37 and CDR3 with the amino acid sequence of SEQ ID NO: 42;
(viii) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 4, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 20, and (b) a light chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 33, CDR2 with the amino acid sequence of SEQ ID NO: 36 and CDR3 with the amino acid sequence of SEQ ID NO: 42;
(ix) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 5, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 21; and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 33, CDR2 with the amino acid sequence of SEQ ID NO: 36 and CDR3 with the amino acid sequence of SEQ ID NO: 42;
(x) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 1, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 22; and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 33, CDR2 with the amino acid sequence of SEQ ID NO: 36 and CDR3 with the amino acid sequence of SEQ ID NO: 42;
(xi) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 6, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 23; and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 33, CDR2 with the amino acid sequence of SEQ ID NO: 36 and CDR3 with the amino acid sequence of SEQ ID NO: 42;
(xii) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 1, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 17; and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 31, CDR2 with the amino acid sequence of SEQ ID NO: 38 and CDR3 with the amino acid sequence of SEQ ID NO: 40;
(xiii)(a) a heavy chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 362, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 363; and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 32, CDR2 with the amino acid sequence of SEQ ID NO: 35 and CDR3 with the amino acid sequence of SEQ ID NO: 41;
(xiv) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 7, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 24; and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 32, CDR2 with the amino acid sequence of SEQ ID NO: 35 and CDR3 with the amino acid sequence of SEQ ID NO: 41;
(xv) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 8, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 25, and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 32, CDR2 with the amino acid sequence of SEQ ID NO: 35 and CDR3 with the amino acid sequence of SEQ ID NO: 41;
(xvi) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 9, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 21; and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 32, CDR2 with the amino acid sequence of SEQ ID NO: 35 and CDR3 with the amino acid sequence of SEQ ID NO: 41; (xvii) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 8, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 26; and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 32, CDR2 with the amino acid sequence of SEQ ID NO: 35 and CDR3 with the amino acid sequence of SEQ ID NO: 41; (xviii) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 8, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 27; and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 32, CDR2 with the amino acid sequence of SEQ ID NO: 35 and CDR3 with the amino acid sequence of SEQ ID NO: 41; (xix) (a) a heavy chain variable domain comprising: CDR1 with the amino acid sequence of SEQ ID NO: 1, CDR2 with the amino acid sequence of SEQ ID NO: 11 and CDR3 with the amino acid sequence of SEQ ID NO: 29; and (b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 31, CDR2 with the amino acid sequence of SEQ ID NO: 34 and CDR3 with the amino acid sequence of SEQ ID NO: 40;
(xx) (a) a heavy chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 8,
CDR2 with the amino acid sequence of SEQ ID NO: 11 and
CDR3 with the amino acid sequence of SEQ ID NO: 30; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 32;
CDR2 with the amino acid sequence of SEQ ID NO: 35 and
CDR3 with the amino acid sequence of SEQ ID NO: 41;
(xxi) CDR1 with the amino acid sequence of SEQ ID NO: 1,
CDR2 with the amino acid sequence of SEQ ID NO: 11 and
CDR3 with the amino acid sequence of SEQ ID NO: 17; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 31,
CDR2 with the amino acid sequence of SEQ ID NO: 34 and
CDR3 with the amino acid sequence of SEQ ID NO: 43; or
(xxii) (a) a heavy chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 1,
CDR2 with the amino acid sequence of SEQ ID NO: 11 and
CDR3 with the amino acid sequence of SEQ ID NO: 17; and
(b) a light chain variable domain comprising:
CDR1 with the amino acid sequence of SEQ ID NO: 31,
CDR2 with the amino acid sequence of SEQ ID NO: 39 and
CDR3 with the amino acid sequence of SEQ ID NO: 40.
10. The isolated monoclonal antibody or antigen -binding fragment thereof according to claim 1 , wherein the heavy chain variable domain comprises an amino acid sequence selected from the group: SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 366, SEQ ID NO: 368, SEQ ID NO: 370 or SEQ ID NO: 372.
11. The isolated monoclonal antibody or antigen -binding fragment thereof according to claim 1 , wherein the light chain variable domain comprises an amino acid sequence selected from the group: SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86 or SEQ ID NO: 87.
12. The isolated monoclonal antibody or antigen -binding fragment thereof according to claim 1, wherein:
(a) the heavy chain variable domain comprises an amino acid sequence selected from the group: SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 366, SEQ ID NO: 368, SEQ ID NO: 370 or SEQ ID NO: 372; and
(b) the light chain variable domain comprises an amino acid sequence selected from the group: SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86 or SEQ ID NO: 87.
13. The isolated monoclonal antibody or antigen -binding fragment thereof according to claim 12, wherein:
(i) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 44 and (b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74;
(ii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 45 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74;
(iii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 46 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 75;
(iv) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 47 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74;
(v) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 48 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 75;
(vi) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 49 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74;
(vii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 50 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74;
(viii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 51 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74; (ix) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 52 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74;
(x) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 53 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74;
(xi) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 54 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 76;
(xii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 55 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 77;
(xiii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 56 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 76;
(xiv) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 57 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 76;
(xv) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 58 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 76;
(xvi) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 59 and (b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 76;
(xvii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 53 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 78;
(xviii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 60 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 75;
(xix) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 61 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 75;
(xx) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 62 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 75;
(xxi) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 63;
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 75;
(xxii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 64 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 75;
(xxiii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 65 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74; (xxiv) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 66 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 75;
(xxv) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 67 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74;
(xxvi) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 53 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 79;
(xxvii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 68 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 75;
(xxviii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 69 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 80;
(xxix) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 70 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 81;
(xxx) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 71 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 75;
(xxxi) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 72 and (b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 82;
(xxxii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 73 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74;
(xxxiii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 53 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 83;
(xxxiv) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 53 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 84;
(xxxv) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 53 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 85;
(xxxvi) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 53 and
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 86;
(xxxvii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 53;
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 87;
(xxxviii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 366;
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74; (xxxix) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 368;
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74;
(xxxx) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 370;
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74; or
(xxxxi) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 372;
(b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 74.
14. The isolated monoclonal antibody according to any one of claims 1 to 13, wherein the antibody that specifically binds to IL-4Rα is a full-length IgG antibody.
15. The isolated monoclonal antibody according to claim 14, wherein the full-length IgG antibody is of human IgGl, IgG2, IgG3 or IgG4 isotype.
16. The isolated monoclonal antibody according to claim 15, wherein the antibody comprises the mutation S228P in the hinge region.
17. The isolated monoclonal antibody according to claim 1, comprising a heavy chain comprising an amino acid sequence selected from the group: SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 367, SEQ ID NO: 369, SEQ ID NO: 371 or , SEQ ID NO: 373.
18. The isolated monoclonal antibody according to claim 1, comprising a light chain comprising an amino acid sequence selected from the group: SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130 or SEQ ID NO: 131.
19. The isolated monoclonal antibody according to claim 1, comprising:
(i) (a) a heavy chain comprising an amino acid sequence that is selected from the group: SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 367, SEQ ID NO: 369, SEQ ID NO: 371 or , SEQ ID NO: 373, and
(b) a light chain comprising an amino acid sequence that is selected from the group: SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130 or SEQ ID NO: 131;
(ii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 88, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(iii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 89, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(iv) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 90, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 119;
(v) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 91, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 118; (vi) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 92, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 119;
(vii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 93, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(viii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 94, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(ix) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 95, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118,
(x) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 96, and
(b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(xi) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(xii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 98, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 120,
(xiii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 99, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 121;
(xiv) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 100, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 120;
(xv) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 101, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 120;
(xvi) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 102, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 120,
(xvii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 103, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 120;
(xviii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 122;
(xix) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 104, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 119;
(xx) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 105, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 119; (xxi) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 106, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 119;
(xxii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 107, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 119;
(xxiii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 108, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 119;
(xxiv) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 109, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(xxv) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 110, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 119;
(xxvi) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 111, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(xxvii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 123;
(xxviii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 112, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 119;
(xxix) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 113, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 124;
(xxx) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 114, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 125;
(xxxi)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 115, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 119;
(xxxii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 116, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 126;
(xxxiii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 117, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(xxxiv) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 127;
(xxxv) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 128; (xxxvi) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 129;
(xxxvii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 130;
(xxxviii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 97, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 131;
(xxxix) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 367, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(xxxx) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 369, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 118;
(xxxxi) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 371, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 11 8; or (xxxxii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 373, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 118.
20. The isolated nucleic acid that encodes the antibody or antigen -binding fragment thereof according to any one of claims 1 to 19.
21. The isolated nucleic acid according to claim 20, wherein the nucleic acid is DNA.
22. An expression vector comprising the nucleic acid according to any one of claims 20 to 21.
23. A method for producing a host cell to produce the antibody or antigen- binding fragment thereof according to any one of claims 1 to 19, comprising cell transformation by the expression vector according to claim 22.
24. A host cell for producing the antibody or antigen -binding fragment thereof according to any one of claims 1 to 19, comprising the nucleic acid according to any one of claims 20 to 21.
25. A method for producing the antibody or antigen -binding fragment thereof according to any one of claims 1 to 19, comprising culturing the host cell according to claim 24 in a growth medium under conditions sufficient to produce said antibody, if necessary, followed by isolation and purification of the resulting antibody.
26. A pharmaceutical composition comprising the antibody or antigen- binding fragment thereof according to any one of claims 1 to 19 in combination with one or more pharmaceutically acceptable excipients.
27. A pharmaceutical composition, comprising the antibody or antigen- binding fragment thereof according to any one of claims 1 to 19 and at least one other therapeutically active compound.
28. The pharmaceutical composition according to claim 27, wherein the other therapeutically active compound is an antibody, a small molecule, a hormone therapy agent or combination thereof.
29. A method for treating a disease or disorder mediated by IL-4Rα, comprising administering to a subject in need of such treatment the antibody or antigen-binding fragment thereof according to any one of claims 1 to 19 or the pharmaceutical composition according to any one of claims 26 to 28 in a therapeutically effective amount.
30. The method for treating a disease or disorder mediated by IL-4Rα according to claim 29, comprising administering to a subject in need of such treatment the antibody or antigen-binding fragment thereof according to any one of claims 1 to 19 or the pharmaceutical composition according to claim 26 and at least one other therapeutically active compound in a therapeutically effective amount.
31. The method for treating a disease or disorder according to claim 30, wherein the other therapeutically active compound is an antibody, a small molecule, a hormone therapy agent or combination thereof.
32. The method for treatment of a disease or disorder according to any one of claims 29-31, wherein the disease or disorder mediated by IL-4Rα is selected from the group: atopic dermatitis, eczema, allergic dermatitis, atopic keratoconjunctivitis, bronchial asthma, chronic obstructive pulmonary disease, allergic bronchopulmonary aspergillosis, chronic rhinosinusitis with nasal polyps, chronic rhinosinusitis, fungal rhinosinusitis, nasal polyps, sleep apnea, allergic rhinitis, Churg-Strauss syndrome, itching associated with chronic liver diseases, hypertrophic scar, alopecia areata, urticaria, allergic dermatitis, focal scleroderma, pemphigus, Netherton syndrome, ichthyosis, eosinophilic gastritis, eosinophilic esophagitis, prurigo nodularis, colitis, filariasis, leishmaniasis, schistosomiasis, arterial hypertension, interstitial lung diseases, metastatic non-small cell lung cancer, metastases of epithelial tumors, malignant neoplasm of the prostate gland, malignant neoplasm of the brain, stomach cancer, colorectal cancer, malignant neoplasm of the colon, malignant neoplasm of the breast, pancreatic cancer, malignant neoplasm of the extrahepatic bile ducts, intrahepatic bile duct cancer, malignant neoplasm of the gallbladder, Hodgkin lymphoma, medulloblastoma, hypereosinophilic syndrome, allergic reactions, acute respiratory distress syndrome.
33. Use of the antibody or antigen -binding fragment thereof according to any one of claims 1 to 19 or the pharmaceutical composition according to any one of claims 26 to 28 for treating a disease or disorder mediated by IL-4Rα .
34. The use according to claim 33, wherein use of the antibody or antigen- binding fragment thereof as claimed in any one of claims 1 to 19 or the pharmaceutical composition according to claim 26 and at least one other therapeutically active compound for the treatment of a disease or disorder mediated by IL-4Rα .
35. The use according to claim 34, wherein the other therapeutically active compound is a small molecule, hormone therapy agent, or any combination thereof.
36. The use according to any one of claims 33-35, wherein the disease or disorder mediated by IL-4Rα is selected from the group: atopic dermatitis, eczema, allergic dermatitis, atopic keratoconjunctivitis, bronchial asthma, chronic obstructive pulmonary disease, allergic bronchopulmonary aspergillosis, chronic rhinosinusitis with nasal polyps, chronic rhinosinusitis, fungal rhinosinusitis, nasal polyps, sleep apnea, allergic rhinitis, Churg-Strauss syndrome, itching associated with chronic liver diseases, hypertrophic scar, alopecia areata, urticaria, allergic dermatitis, focal scleroderma, pemphigus, Netherton syndrome, ichthyosis, eosinophilic gastritis, eosinophilic esophagitis, prurigo nodularis, colitis, filariasis, leishmaniasis, schistosomiasis, arterial hypertension, interstitial lung diseases, metastatic non-small cell lung cancer, metastases of epithelial tumors, malignant neoplasm of the prostate gland, malignant neoplasm of the brain, stomach cancer, colorectal cancer, malignant neoplasm of the colon, malignant neoplasm of the breast, pancreatic cancer, malignant neoplasm of the extrahepatic bile ducts, intrahepatic bile duct cancer, malignant neoplasm of the gallbladder, Hodgkin lymphoma, medulloblastoma, hypereosinophilic syndrome, allergic reactions, acute respiratory distress syndrome.
PCT/RU2022/050351 2021-11-11 2022-11-07 Monoclonal antibody or antigen-binding fragment thereof that specifically binds to il-4ra, and use thereof WO2023085978A1 (en)

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Citations (4)

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WO2001092340A2 (en) * 2000-05-26 2001-12-06 Immunex Corporation Use of interleukin-4 antagonists and compositions thereof
WO2005047331A2 (en) * 2003-11-07 2005-05-26 Immunex Corporation Antibodies that bind interleukin-4 receptor
WO2010053751A1 (en) * 2008-10-29 2010-05-14 Regeneron Pharmaceuticals, Inc. High affinity human antibodies to human il-4 receptor
WO2019148405A1 (en) * 2018-02-01 2019-08-08 北京凯因科技股份有限公司 IL-4Rα ANTIBODY AND USE THEREOF

Patent Citations (4)

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Publication number Priority date Publication date Assignee Title
WO2001092340A2 (en) * 2000-05-26 2001-12-06 Immunex Corporation Use of interleukin-4 antagonists and compositions thereof
WO2005047331A2 (en) * 2003-11-07 2005-05-26 Immunex Corporation Antibodies that bind interleukin-4 receptor
WO2010053751A1 (en) * 2008-10-29 2010-05-14 Regeneron Pharmaceuticals, Inc. High affinity human antibodies to human il-4 receptor
WO2019148405A1 (en) * 2018-02-01 2019-08-08 北京凯因科技股份有限公司 IL-4Rα ANTIBODY AND USE THEREOF

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