Anti-CD79 antibody and application thereof

Abstract

The present disclosure provides humanized and affinity mature antibodies (mAbs) and fragments thereof that specifically bind CD79 with high affinity. Anti-CD79mAb and fragments thereof are useful for the treatment of antibody-related disorders, including autoimmune diseases, allergy, transplant rejection, or immune-mediated rejection of therapeutic agents, optionally in combination with additional therapeutic agents. Furthermore, anti-CD79mAb and fragments thereof can be used for diagnostic purposes, including detection of CD79 cells in a biological sample and diagnosis of B-cell related diseases.

Description

Statement Regarding Federally Funded Research

[0001] This invention was made with government support under Grant No. 1R43AI120433-01 awarded by the National Institute of Allergy and Infectious Diseases (NIAID). The government has certain rights in this invention. Reference to a sequence listing, table or computer program

[0002] The formal text of the sequence listing is submitted together with this specification through EFS-Web in an ASCII text file, with the file name "PRI011_ST25.txt", the creation date of which is September 30, 2019, and the size is 73 kilobytes. The sequence listing submitted through EFS-Web is part of this specification and is hereby incorporated by reference in its entirety. Background Art

[0003] B cells play a major role in the pathogenesis of many autoimmune diseases, including rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), multiple sclerosis, and type 1 diabetes (T1D), as demonstrated by the efficacy of B cell-directed therapies, such as the use of rituximab in these diseases. Unfortunately, current therapies are based on B cell depletion, which is problematic from a safety perspective. Existing standard of care therapies can have adverse effects, particularly opportunistic infections and latent viral reactivation, due to prolonged and severe B cell depletion, due to the consequent immunosuppression.

[0004] CD79 (Cluster of Differentiation 79) is a transmembrane protein that forms a complex with the B-cell receptor (BCR) and generates signals upon antigen recognition by the BCR. CD79 consists of two distinct chains, called CD79A and CD79B (formerly Ig-α and Ig-β); they form heterodimers on the surface of B cells stabilized by disulfide bonds. Both CD79a and CD79b are members of the immunoglobulin superfamily. CD79 has been used as a pan-B cell marker and can be used to detect B-cell neoplasms / tumors.

[0005] Unlike anti-CD20 mAbs, the protective effects of mAbs directed against CD79 do not require cell depletion; instead, they act by inducing a reversible anergy or anergy state and therefore do not participate in the generation of an immune response. In animal models, anti-CD79 antibodies provide immunosuppression and reduce inflammation by inducing B cell anergy. Summary of the invention

[0006] The present disclosure provides antibodies that specifically bind to CD79 with high affinity, including antibody fragments thereof. The antibody can be a monoclonal antibody, and can be a chimeric or humanized antibody. Chimeric anti-CD79 antibodies including fragments thereof can have non-human (e.g., mouse) complementary determining regions (CDRs) and non-human framework regions, and optionally one or more human constant regions. Non-human, heavy chain and light chain variable regions include SEQ ID NOs: 1-2. Humanized anti-CD79 antibodies including fragments thereof can have non-human (e.g., mouse) CDRs and human framework regions, and optionally non-human framework amino acid residues adjacent to CDRs and optionally one or more human constant regions. Non-human CDRs include, for example, VH CDR1-3 of SEQ ID NOs: 3-5 and VL-CDR1-3 of SEQ ID NOs: 6-8.

[0007] The disclosed humanized antibodies represent anti-CD79 antibodies obtained by transplanting the CDRs of SEQ ID NO:3-8 to the human framework of the heavy chain and the human framework of the light chain, together with a selected number of framework residues from mouse antibodies. Nine variable regions (SEQ ID NO:9-17) for humanized heavy chains and six variable regions (SEQ ID NO:18-23) for humanized light chains can be combined to prepare anti-CD79 humanized antibodies. Anti-CD79 antibodies disclosed herein also include those obtained from affinity maturation libraries made from humanized anti-CD79 antibodies. Anti-CD79 antibodies can be prepared with a heavy chain variable region selected from SEQ ID NO:9-17, 24-27, 32-41, 71, 72 and 75-77 and a light chain variable region selected from SEQ ID NO:18-23, 28-31, 42-56 and 73-74. The anti-CD79 antibody may also include a heavy chain variable region having 99%, 95%, 90%, 80% or 70% sequence identity to one of SEQ ID NOs: 9-17, 24-27, 32-41, 71, 72 and 75-77, and a light chain variable region having 99%, 95%, 90%, 80% or 70% sequence identity to one of SEQ ID NOs: 18-23, 28-31, 42-56 and 73-74. The anti-CD79 antibody may bind to CD79 with an affinity of 2.0-5.1 nM, or 45 nM to 300 nM, or 2.0 to 300 nM. The anti-CD79 antibody may bind with an affinity of at least 300 nM, or at least 140 nM, or at least 100 nM, or at least 5.1 nm, at least 3.8 nM, or at least 2.4 nM.

[0008] The anti-CD79 antibodies described herein may include modifications that provide the antibodies with desired properties. For example, the modifications may increase the serum half-life of the antibody, or the modifications may reduce the serum half-life. The modifications may also increase or reduce the effector function of the antibody. The modifications may reduce immunogenicity, or reduce other unwanted side effects or adverse events caused by the anti-CD79 antibodies.

[0009] The anti-CD79 antibodies described herein can induce an anergy in the B-cells of the subject and can therefore be used to treat certain autoimmune diseases. For example, autoimmune diseases associated with anti-autoantibody reactions can be treated with anti-CD79 antibodies because the induced anergy will prevent the production of anti-autoantibodies. The anti-CD79 antibodies described herein can also be used to induce an anergy in any situation with an undesirable antibody response. The anti-CD79 antibodies described herein can be used to induce an anergy in B cells. The anti-CD79 antibodies described herein can be used to inhibit the proliferation of B cells. The anti-CD79 antibodies described herein can be used to prevent infusion reactions caused by pre-existing anti-drug antibodies. The anti-CD79 antibodies described herein can be used to prevent the formation of anti-drug antibodies after injection of biological therapy. Undesirable antibody-related conditions may include, for example, autoimmune diseases, certain allergies (allergies associated with antibodies), certain type I diabetes, etc. Autoimmune diseases that can be treated with anti-CD79 antibodies include, for example, systemic lupus erythematosus (SLE), inflammatory bowel disease (such as Crohn's disease and ulcerative colitis), rheumatoid arthritis, multiple sclerosis, Grave's disease, CREST syndrome, systemic sclerosis, celiac disease, etc. Other autoimmune diseases include,Examples include achalasia, Addison's disease, adult-onset Still's disease, agammaglobulinemia, alopecia areata, amyloidosis, ankylosing spondylitis, anti-GBM / anti-TBM nephritis, antiphospholipid syndrome, autoimmune angioedema, autoimmune dysautonomia, autoimmune encephalomyelitis, autoimmune hepatitis, autoimmune inner ear disease (AIED), autoimmune myocarditis, autoimmune oophoritis, autoimmune orchitis, autoimmune pancreatitis, autoimmune retinopathy, autoimmune urticaria, axonal and neuronal neuropathy (AMAN), Baló disease, Behcet's disease, benign mucous membrane pemphigoid, bullous pemphigoid, Castleman disease (CD), celiac disease, Chagas disease, chronic inflammatory demyelinating polyneuritis (CIDP), chronic recurrent multifocal osteomyelitis (CRMO), Churg-Strauss syndrome (CSS) or eosinophilic granulomatosis (EGPA), cicatricial pemphigoid, Cogan's syndrome, cold agglutinin disease, congenital heart block, Coxsackie myocarditis, CREST syndrome, Crohn's Crohn's disease, dermatitis herpetiformis, dermatomyositis, Devic's disease (neuromyelitis optica), discoid lupus, Dressler's syndrome, endometriosis, eosinophilic esophagitis (EoE), eosinophilic fasciitis, erythema nodosum, essential mixed cryoglobulinemia, Evans syndrome, fibromyalgia, fibrosing alveolitis, giant cell arteritis (temporal arteritis), giant cell myocarditis, glomerulonephritis, Goodpasture's syndrome, granulomatosis with polyangiitis, Graves' disease, Guillain-Barré syndrome e syndrome, Hashimoto's thyroiditis, hemolytic anemia, Henoch-Schonlein purpura (HSP), herpes gestationis or pemphigoid gestationis (PG), hidradenitis suppurativa (HS) (acne inversa), hypogammaglobulinemia, IgA nephropathy, IgG4-related sclerosing disease, immune thrombocytopenic purpura (ITP), inclusion body myositis (IBM), interstitial cystitis (IC), juvenile arthritis, juvenile diabetes mellitus (type 1 diabetes), juvenile myositis (JM), Kawasaki disease,Lambert-Eaton syndrome, leukocytoclastic vasculitis, lichen planus, lichen sclerosus, woody conjunctivitis, linear IgA disease (LAD), lupus, chronic Lyme disease, Meniere's disease, microscopic polyangiitis (MPA), mixed connective tissue disease (MCTD), Mooren's ulcer, Mucha-Habermann disease, multifocal motor neuropathy (MMN) or MMNCB, multiple sclerosis, myasthenia gravis, myositis, narcolepsy, neonatal lupus, neuromyelitis optica, neutropenia, ocular cicatricial pemphigoid, optic neuritis, relapsing rheumatic disease (PR), PANDAS, paraneoplastic cerebellar degeneration (PCD), paroxysmal nocturnal hemoglobinuria (PNH), Parry's Romberg syndrome, pars planitis (peripheral uveitis), Parsonage-Turner syndrome, pemphigus, peripheral neuropathy, peripheral encephalomyelitis, pernicious anemia (PA), POEMS syndrome, polyarteritis nodosa, polyglandular syndromes (type I, II, III), polymyalgia rheumatica, polymyositis, post-myocardial infarction syndrome, postpericardiotomy syndrome, primary biliary cirrhosis, primary sclerosing cholangitis, progesterone dermatitis, psoriasis, psoriatic arthritis, pure red cell agenesis (PRCA), pyoderma gangrenosum, Raynaud's phenomenon, reactive arthritis, reflex sympathetic dystrophy, relapsing polychondritis, restless legs syndrome (RLS), retroperitoneal fibrosis, rheumatic fever, rheumatoid arthritis, sarcoidosis, Schmidt syndrome, scleritis, scleroderma, Syndrome, sperm and testicular autoimmunity, stiff-man syndrome (SPS), subacute bacterial endocarditis (SBE), Susac's syndrome, sympathetic ophthalmia (SO), Takayasu's arteritis, temporal arteritis / giant cell arteritis, thrombocytopenic purpura (TTP), Tolosa-Hunt syndrome (THS), transverse myelitis, type 1 diabetes, ulcerative colitis (UC), undifferentiated connective tissue disease (UCTD), uveitis, vasculitis, vitiligo, Vogt-Koyanagi-Harada disease. These undesirable antibody-related disorders can be treated by administering one or more anti-CD79 antibodies described herein to a subject suffering from an undesirable antibody-related disorder.

[0010] Anti-CD79 antibodies described herein can be used to treat CD79-positive hematopoietic cancers, such as lymphomas and leukemias. They can also be used in chimeric antigen receptors (CARs) to make immune cells with anti-CD79CARs. These anti-CD79CAR immune cells include, for example, T-cells or natural killer cells with anti-CD79 CARs. Anti-CD79 CAR T-cells and / or natural killer cells can be used to treat diseases in which cells that cause diseases display CD79. The diseases include, for example, CD79-positive hematopoietic cancers, such as lymphomas and leukemias. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] A better understanding of the features and advantages of the present disclosure will be obtained by referring to the following detailed description, which sets forth illustrative embodiments of the present disclosure and the accompanying drawings.

[0012] Figure 1 is a graph showing the development of type 1 diabetes over time in VH125NOD mice.

[0013] Figure 2 is a graph showing the binding of soluble CD79 antigen to cells expressing anti-CD79 CAR-T constructs.

[0014] Figure 3a Competitive binding between antibodies LB517 / LB519 (hCur14 FALA) and Curly-14 for binding to B cells is shown. Figure 3b Shown is PTEN in B-cells treated with different anti-hCD79 antibodies. Figure 3c Shown is the B-cell receptor (BCR) expression after treatment of B-cells with different anti-hCD79 antibodies.

[0015] Figure 4a Shown is calcium influx in B-cells after treatment with different anti-hCD79 antibodies. Figure 4b Desensitization of the B-cell receptor after treatment of B-cells with different anti-hCD79 antibodies is shown.

[0016] Figure 5 Shown is the effect of treatment with anti-hCD79 antibodies on the development of arthritis in an arthritis model system.

[0017] Figure 6 The efficacy of treatment with anti-hCD79 antibodies on lupus development in a lupus model system is shown.

[0018] Figure 7 The efficacy of treatment with anti-hCD79 antibodies on the progression of multiple sclerosis in a model system of MS is shown. DETAILED DESCRIPTION

[0019] Although various embodiments of the present disclosure are described herein, it will be apparent to those skilled in the art that these embodiments are provided by way of example only. Without departing from the present disclosure, various modifications and changes and variations and substitutions of the embodiments described herein will be apparent to those skilled in the art. It should be understood that various alternatives to the embodiments described herein may be adopted in practicing the present invention. It should also be understood that each embodiment of the present disclosure may be optionally combined with any one or more other embodiments described herein that are consistent with the embodiment.

[0020] Where elements are presented in list format (eg, in Markush groups), it is understood that every possible subgroup of the elements is also disclosed, and any one or more elements may be removed from the list or group.

[0021] It should also be understood that, unless explicitly indicated to the contrary, in any method described or claimed herein that includes more than one act or step, the order of the acts or steps of the method is not necessarily limited to the order in which the acts or steps of the method are recorded, but the present disclosure covers embodiments of the order that are so limited.

[0022] It should also be understood that, generally, where an embodiment in the specification or claims is referred to as comprising one or more features, the disclosure also covers embodiments consisting of or consisting essentially of the described features.

[0023] It should also be understood that any embodiment of the present disclosure, such as any embodiment found within the prior art, may be explicitly excluded from the claims, regardless of whether a specific exclusion is recited in the specification.

[0024] It should also be understood that the peptides, polypeptides or proteins, such as antibodies or fragments thereof, mentioned herein include pharmaceutically acceptable salts thereof, unless otherwise expressly stated or the context clearly indicates otherwise. Such salts may have a positive net charge, a negative net charge or no net charge.

[0025] Headings are included herein for reference and to aid in locating certain sections. Headings are not intended to limit the scope of the embodiments and concepts described in the sections under those headings, and those embodiments and concepts may be applied in other sections throughout the entire disclosure.

[0026] All patent documents and all non-patent documents cited herein are incorporated by reference in their entirety to the same extent as if each individual patent document or non-patent document was specifically and individually indicated to be incorporated by reference in its entirety. definition

[0027] Unless otherwise defined or clearly indicated otherwise by usage herein, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

[0028] As used in the specification and the appended claims, the indefinite articles "a" and "an" and the definite article "the" may include plural as well as singular referents, unless expressly stated otherwise or the context clearly dictates otherwise.

[0029] The term "about" or "approximately" represents an acceptable error for a particular value determined by a person of ordinary skill in the art, which depends in part on how the value is measured or determined. In certain embodiments, the term "about" or "approximately" means within one standard deviation. In certain embodiments, when a specific error range (e.g., the standard deviation of the mean value given in a chart or data table) is not recorded, the term "about" or "approximately" means that the range of the value will be covered, and considering the important figures, the range that can also be included by rounding up or down to the value. In certain embodiments, the term "about" or "approximately" refers to within ±10%, 5%, 4%, 3%, 2% or 1% of the specified value. Whenever the term "about" or "approximately" is located before the first value in a series of two or more numerical values ​​or a series of two or more numerical ranges, the term "about" or "approximately" applies to each of the numerical values ​​in the series or the numerical value in the series.

[0030] The term "antibody" refers to a protein that is functionally defined as a binding protein and structurally defined as comprising an amino acid sequence that is recognized as being derived from a framework region of an immunoglobulin encoding gene. An antibody can be composed of one or more polypeptides that are substantially encoded by an immunoglobulin gene or a fragment of an immunoglobulin gene. Recognized immunoglobulin genes include κ, λ, α, γ, δ, ε, and μ constant region genes, as well as countless immunoglobulin variable region genes. Light chains are classified as κ or λ. Heavy chains are classified as γ, μ, α, δ, or ε, which in turn define immunoglobulin classes, which are IgG, IgM, IgA, IgD, and IgE, respectively.

[0031] It is known that the typical gamma immunoglobulin (antibody) structural unit comprises a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one "light" chain (about 25 kD) and one "heavy" chain (about 50-70 kD). The N-terminus of each chain defines a variable region of about 100 to 110 or more amino acids that is primarily responsible for antigen recognition. The term variable light chain (V L ) and variable heavy chain (V H ) refer to these light and heavy chains, respectively.

[0032] Antibodies exist as intact immunoglobulins or as a number of well-characterized fragments. Thus, for example, pepsin digests an antibody below the disulfide bonds in the hinge region to produce F(ab)' 2 , which is a dimer of Fab', which itself is a light chain naturally connected to VH-CH1-hinge by a disulfide bond. F(ab)' can be reduced under mild conditions 2 , to break the disulfide bonds in the hinge region, thereby converting (Fab') 2 The dimer is converted into a Fab' monomer. The Fab' monomer is essentially a Fab with a partial hinge region (see, Fundamental Immunology, WE Paul, ed., Raven Press, NY (1993) for a more detailed description of other antibody fragments). Although various antibody fragments are defined in terms of digestion of intact antibodies, it will be appreciated by those skilled in the art that fragments can be synthesized de novo chemically or by utilizing recombinant DNA methods. Therefore, the term antibody as used herein also includes antibody fragments produced by modifying intact antibodies or synthesized using recombinant DNA methods. Preferred antibodies include V H -V L Dimers include single-chain antibodies (antibodies that exist as a single polypeptide chain), such as single-chain Fv antibodies (sFv or scFv), in which the variable heavy chain and variable light chain regions are linked together (directly or through a peptide linker) to form a continuous polypeptide. Single-chain Fv antibodies are covalently linked V H -V L Heterodimers can be derived from V H - and V L - nucleic acid expression of the coding sequence (e.g., Huston et al., Proc. Nat. Acad. Sci. USA, 85:5879-5883, 1988, which is incorporated herein by reference in its entirety). H and V L As a polypeptide chain connected to each other, V H and V LThe domains are non-covalently bound. Alternatively, the antibody can be another fragment. Other fragments can also be produced, including using recombinant technology. For example, if one of the chains (heavy chain or light chain) is fused to the g3 capsid protein and the complementary chain is exported to the periplasm as a soluble molecule, the Fab molecule can be displayed on the phage. The two chains can be encoded on the same or different replicons; the two antibody chains in each Fab molecule will be assembled after translation, and then the dimer will be incorporated into the phage particle by connecting to one chain of g3p (see, for example, U.S. Patent No.: 5,733,743, which is incorporated herein by reference in its entirety). ScFv antibodies and multiple other structures convert naturally aggregated but chemically separated light and heavy polypeptide chains from antibody V regions into molecules folded into a three-dimensional structure, which is substantially similar to the structure of the antigen binding site, which is known to those skilled in the art (see, for example, U.S. Patent Nos. 5,091,513, 5,132,405 and 4,956,778, all of which are incorporated herein by reference in their entirety). Particularly preferred antibodies include all those that have been displayed on phage or produced by recombinant technology using vectors in which the chains are secreted in the form of soluble proteins, such as scFv, Fv, Fab, (Fab') 2 Antibodies can also include diabodies and minibodies.

[0033] Antibodies also include heavy chain dimers, such as antibodies from camelids. H The V region of the heavy chain does not have to make hydrophobic interactions with the light chain, so the region in the heavy chain that normally contacts the light chain is changed to hydrophilic amino acid residues in camelids. H Domain V HH Structural domain.

[0034] In camelids, the diversity of the antibody repertoire depends on V H or V HH The complementarity determining regions (CDRs) 1, 2 and 3 in the camel V HH The CDR3 in the V region is characterized by its relatively long length, averaging 16 amino acids (Muyldermans et al., 1994, Protein Engineering 7(9):1129, which is incorporated herein by reference in its entirety). This is in contrast to the CDR3 regions of antibodies from many other species. For example, the mouse V H The CDR3 of a gene has an average of 9 amino acids.

[0035] A library of camelid-derived antibody variable regions that maintains the in vivo diversity of camelid variable regions can be prepared by, for example, the method disclosed in U.S. Patent Application Publication No. US20050037421, published on February 17, 2005, which is incorporated herein by reference in its entirety.

[0036] As used herein, the term "binding specificity" of an antibody refers to the identity of the antigen to which the antibody binds, preferably the identity of the epitope to which the antibody binds.

[0037] As used herein, the term "chimeric polynucleotide" refers to a polynucleotide comprising a wild-type region and a mutant region. It may also refer to a polynucleotide comprising a wild-type region from one polynucleotide and a wild-type region from another related polynucleotide.

[0038] As used herein, the term "complementarity determining region" or "CDR" refers to the art-recognized term exemplified by Kabat and Chothia. CDRs are also often referred to as hypervariable regions or hypervariable loops (Chothia and Lesk (1987) J Mol. Biol. 196: 901; Chothia et al., (1989) Nature 342: 877; EA Kabat et al., Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md.) (1987); and Tramontano et al., (1990) J Mol. Biol. 215: 175, all of which are incorporated herein by reference in their entirety). "Framework region" or "FR" refers to the region flanking the CDRs of a V domain. The positions of CDRs and framework regions can be determined using various definitions well known in the art, such as Kabat, Chothia, the International ImMunoGeneTics database (IMGT), and AbM (see, e.g., Johnson et al., supra; Chothia & Lesk, 1987, Canonical structures for the hypervariable regions of immunoglobulins. J. Mol. Biol. 196, 901-917; Chothia C. et al., 1989, Conformations of immunoglobulin hypervariable regions. Nature 342, 877-883; Chothia C. et al., 1992, structural repertoire of the human VH segments J. Mol. Biol. 227, 799-817; Al-Lazikani et al., J. Mol. Biol 1997, 273 (4)). The definition of antigen binding sites is also described in Ruiz et al., IMGT, the international ImMunoGeneTics database.Nucleic Acids Res., 28, 219-221 (2000); and Lefranc, M.-P. IMGT, the international ImMunoGeneTics database. Nucleic Acids Res., Jan 1; 29(l): 207-9 (2001); MacCallum et al., Antibody-antigen interactions: Contact analysis and binding site topography, J. Mol. Biol., 262(5), 732-745 (1996); and Martin et al., Proc. Natl Acad. Sci. USA, 86, 9268-9272 (1989); Martin et al., Methods Enzymol., 203, 121-153, (1991); Pedersen et al., Immunomethods, 1, 126, (1992); and Rees et al., In Sternberg MJE (ed.), Protein Structure Prediction. Oxford University Press, Oxford, 141-172, 1996, both of which are incorporated herein by reference in their entirety).

[0039] Whenever the term "at least" or "greater than" precedes the first value in a series of two or more values, the term "at least" or "greater than" applies to every value in the series.

[0040] The term "heterologous" refers to an amino acid or nucleotide sequence that is not found naturally in the context of the amino acid or nucleotide sequence to which it is related.

[0041] Whenever the term "not more than" or "less than" precedes the first value in a series of two or more values, the term "not more than" or "less than" applies to every value in the series of values.

[0042] The term "polynucleotide" refers to a polymer consisting of nucleotide units. Polynucleotides include naturally occurring nucleic acids, such as deoxyribonucleic acid ("DNA") and ribonucleic acid ("RNA"), and nucleic acid analogs. Nucleic acid analogs include those comprising non-naturally occurring bases, nucleotides bonded to other nucleotides other than naturally occurring phosphodiester bonds, or / and bases connected by bonds other than phosphodiester bonds. Non-limiting examples of nucleotide analogs include thiophosphates, dithiophosphates, phosphotriesters, phosphoramides, boric acid phosphates, methylphosphonates, chiral methylphosphonates, 2-O-methyl ribonucleotides, peptide nucleic acids (PNA), etc. Such polynucleotides can be synthesized, for example, using an automated DNA synthesizer. The term "nucleic acid molecule" generally refers to larger polynucleotides. The term "oligonucleotide" generally refers to shorter polynucleotides. In certain embodiments, an oligonucleotide comprises no more than about 50 nucleotides. It should be understood that when a nucleotide sequence is represented by a DNA sequence (ie, A, T, G, C), it also includes an RNA sequence (ie, A, U, G, C) in which "U" replaces "T".

[0043] The term "polypeptide" refers to a polymer composed of natural or / and non-natural amino acid residues, naturally occurring structural variants thereof, or / and synthetic non-naturally occurring analogs thereof connected by peptide bonds. Synthetic polypeptides can be synthesized, for example, using an automated polypeptide synthesizer. Polypeptides can also be recombinantly produced in cells expressing nucleic acid sequences encoding polypeptides. The term "protein" generally refers to larger polypeptides. The term "peptide" generally refers to shorter polypeptides. In certain embodiments, a peptide comprises no more than about 50, 40, or 30 amino acid residues. Polypeptides include antibodies and fragments thereof. Conventional symbols are used herein to depict polypeptide sequences: the left-hand end of a polypeptide sequence is the amino (N)-terminus; the right-hand end of a polypeptide sequence is the carboxyl (C)-terminus.

[0044] The polypeptide may include one or more modifications that may be made during the synthesis or cellular production of the polypeptide, such as one or more post-translational modifications, whether or not the one or more modifications are intentional. Modifications may include, but are not limited to, glycosylation (e.g., N-linked glycosylation and O-linked glycosylation), lipidation, oxidative phosphorylation, sulfation, acetylation (e.g., acetylation of the N-terminus), amidation (e.g., amidation of the C-terminus), hydroxylation, methylation, formation of intramolecular or intermolecular disulfide bonds, formation of lactams between two side chains, formation of pyroglutamic acid, and ubiquitination. As another example, the polypeptide may be linked to a natural polymer (e.g., a polysaccharide) or a synthetic polymer (e.g., polyethylene glycol [PEG]), lipidation (e.g., with C 8 -C 20The polypeptide may be pegylated (acylated by acyl groups) or labeled with a detectable agent (e.g., a radionuclide, a fluorescent dye, or an enzyme). Pegylation can increase protease resistance, stability, and half-life, increase solubility, and reduce aggregation of polypeptides.

[0045] The term "conservative substitution" refers to substitution of an amino acid in a polypeptide with a functionally, structurally or chemically similar natural or non-natural amino acid. In certain embodiments, the following groups each comprise natural amino acids that are conservatively substituted for each other: 1) Glycine (Gly / G), Alanine (Ala / A); 2) Isoleucine (Ile / I), leucine (Leu / L), methionine (Met / M), valine (Val / V); 3) Phenylalanine (Phe / F), tyrosine (Tyr / Y), tryptophan (Trp / W); 4) Serine (Ser / S), threonine (Thr / T), cysteine ​​(Cys / C); 5) Asparagine (Asn / N), glutamine (Gln / Q); 6) Aspartic acid (Asp / D), Glutamic acid (Glu / E); and 7) Arginine (Arg / R), Lysine (Lys / K), Histidine (His / H).

[0046] In further embodiments, the following groups each comprise natural amino acids that are conservative substitutions for one another: 1) Non-polar: Ala, Val, Leu, Ile, Met, Pro (proline / P), Phe, Trp; 2) Hydrophobicity: Val, Leu, Ile, Phe, Tyr, Trp; 3) Aliphatic: Ala, Val, Leu, Ile; 4) Aromatic: Phe, Tyr, Trp, His; 5) uncharged polar or hydrophilic: Gly, Ala, Pro, Ser, Thr, Cys, Asn, Gln, Tyr (tyrosine can be considered a hydrophobic amino acid with a polar side group); 6) Containing aliphatic hydroxyl or sulfhydryl groups: Ser, Thr, Cys; 7) Containing amide: Asn, Gln; 8) Acidic: Asp, Glu; 9) Basic: Lys, Arg, His; and 10) Small: Gly, Ala, Ser, Cys.

[0047] In other embodiments, the amino acids may be listed in groups as follows: 1) Hydrophobicity: Val, Leu, Ile, Met, Phe, Trp, Tyr; 2) Aromatic: Phe, Tyr, Trp, His; 3) Neutral hydrophilic: Gly, Ala, Pro, Ser, Thr, Cys, Asn, Gln; 4) Acidic: Asp, Glu; 5) Basic: Lys, Arg, His; and 6) Residues that affect backbone orientation: Pro, Gly.

[0048] A polypeptide having one or more modifications relative to a parent polypeptide may be appropriately referred to as an "analog," "derivative," or "variant" of the parent polypeptide.

[0049] The present disclosure encompasses pharmaceutically acceptable salts of polypeptides, including those with a positive net charge, those with a negative net charge, and those with no net charge.

[0050] The term "pharmaceutically acceptable" refers to a substance (e.g., active ingredient or excipient) that is suitable for contact with the tissues and organs of a subject without excessive irritation, allergic response, immunogenicity, and toxicity, commensurate with a reasonable benefit / risk ratio, and effective for its intended use. A "pharmaceutically acceptable" excipient or carrier of a pharmaceutical composition is also compatible with the other ingredients of the composition.

[0051] The term "stringent hybridization conditions" refers to hybridization in 50% formamide in 5X SSC at a temperature of 42°C and filter washing in 0.2X SSC at 60°C (1X SSC is 0.15M NaCl, 0.015M sodium citrate). Stringent hybridization conditions also include lower ionic strength and higher washing temperature, such as 0.015M sodium chloride / 0.0015M sodium citrate / 0.1% sodium dodecyl sulfate at 50°C; hybridization with a denaturing agent (e.g., formamide), such as 50% (v / v) formamide at 42°C with 0.1% bovine serum albumin / 0.1% Ficoll / 0.1% polyvinyl pyrrolidone / 50mM sodium phosphate buffer, pH 6.5, with 750mM sodium chloride, 75mM sodium citrate; or 50% formamide, 5X SSC (0.75M NaCl, 0.075M sodium citrate), 50mM sodium phosphate (pH 6.8), 0.1% sodium pyrophosphate, 5X Denhardt's solution, sonicated salmon sperm DNA (50μg / ml), 0.1% SDS and 10% dextran sulfate at 42°C, wash with 0.2X SSC (sodium chloride / sodium citrate) and 50% formamide at 55°C, followed by a high stringency wash at 55°C in 0.1X SSC with EDTA.

[0052] The term "subject" refers to an animal, including but not limited to a mammal, such as a primate (e.g., a human, chimpanzee, or monkey), a rodent (e.g., a rat, a mouse, a guinea pig, a gerbil, or a hamster), a lagomorph (e.g., a rabbit), a porcine (e.g., a pig), an equine (e.g., a horse), a canine (e.g., a dog), or a feline (e.g., a cat).

[0053] In the context of two polypeptides or polynucleotides, the term "substantially homologous" or "substantially identical" refers to two or more sequences or subsequences having at least about 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% amino acid or nucleic acid residue identity when compared and aligned for maximum correspondence using a sequence comparison algorithm or as measured by visual inspection. The term "substantially homologous" or "substantially identical" may mean at least about 70% amino acid or nucleic acid residue identity. The term "substantially homologous" or "substantially identical" may mean at least about 85% amino acid or nucleic acid residue identity. Substantial homology or identity may exist over a sequence region of at least about 20, 30, 40, 50, 100, 150 or 200 residues in length. Sequences may be substantially homologous or identical over the entire length of either or both compared biopolymers.

[0054] Optimal alignment of sequences for comparison can be performed, for example, by the local homology algorithm of Smith and Waterman, Adv. Appl. Math., 2:482 (1981); by the homology alignment algorithm of Needleman and Wunsch, J. Mol. Biol., 48:443 (1970); by the similarity search method of Pearson and Lipman, Proc. Natl. Acad. Sci. USA, 85:2444 (1988); by computerized implementations of these algorithms (e.g., GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computing Group, Madison, Wisconsin, USA); or by visual inspection.

[0055] An example of a useful algorithm is PILEUP. PILEUP creates a multiple sequence alignment from a group of related sequences using progressive pairwise alignments to show relationships and percent sequence identity. It also draws a tree or dendrogram showing the clustering relationships used to create the alignment. PILEUP uses a simplification of the progressive alignment method of Feng and Doolittle, J. Mol. Evol., 35:351-360 (1987). The method used is similar to the method described by Higgins and Sharp, CABIOS, 5:151-153 (1989). The program can align up to about 300 sequences, each with a maximum length of about 5,000 nucleotides or amino acids. The multiple alignment program starts with a pairwise alignment of the two most similar sequences, producing two aligned sequence clusters. The cluster is then aligned with the next most related sequence or aligned sequence cluster. Two sequence clusters can be aligned by a simple extension of the pairwise alignment of two individual sequences. The final alignment is achieved by a series of progressive pairwise alignments. The program is run by specifying specific sequences and their amino acid or nucleotide coordinates for sequence comparison regions and by specifying program parameters. For example, a reference sequence can be compared to other test sequences to determine a percent sequence identity relationship using the following parameters: default gap weight (3.00), default gap length weight (0.10), and weighted end gaps. Another algorithm useful for generating multiple alignments of sequences is Clustal W (see, e.g., Thompson et al., Nucleic Acids Research, 22: 4673-4680

[1994] ).

[0056] Another example of an algorithm suitable for determining percent sequence identity and sequence similarity is the BLAST algorithm, which is described in Altschul et al., J. Mol. Biol., 215: 403-410 (1990). Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information. The algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence that match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is called the neighborhood word score threshold (Altschul 1990). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are then extended in both directions along each sequence for as far as the cumulative alignment score can be increased. For nucleotide sequences, the cumulative score is calculated using the parameters M (reward score for a pair of matching residues; always > 0) and N (penalty score for mismatching residues; always < 0). For amino acid sequences, a scoring matrix is ​​used to calculate the cumulative score. Extension of word hits in each direction is stopped when: the cumulative alignment score falls by the amount X from its maximum achieved value; the cumulative score becomes zero or lower due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses defaults, e.g., a wordlength (W) of 11, an expectation (E) of 10, M=5, N=-4, and a comparison of both strands. For amino acid sequences, the BLASTP program uses defaults, e.g., a wordlength (W) of 3, an expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff and Henikoff, Proc. Natl. Acad. Sci. USA, 89:10915

[1989] ).

[0057] In addition to calculating percent sequence identity, the BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, e.g., Karlin and Altschul, Proc. Natl. Acad. Sci. USA, 90:5873-5787

[1993] ). One measure of similarity provided by the BLAST algorithm is the smallest sum probability [P(N)], which represents the probability that a match between two nucleotide or amino acid sequences would occur by chance. In certain embodiments, a polynucleotide is considered similar to a reference sequence if the smallest sum probability in a comparison of the test polynucleotide to the reference polynucleotide is less than about 0.1, 0.01, or 0.001.

[0058] A polypeptide may be substantially homologous or identical to a second polypeptide if the two polypeptides differ only by conservative amino acid substitutions. As described herein, two nucleic acid sequences may be substantially homologous or identical if the two polynucleotides hybridize to each other under stringent conditions or under highly stringent conditions.

[0059] The term "therapeutically effective amount" refers to an amount of a compound that, when administered to a subject, is sufficient to prevent, reduce the risk of development, delay the onset of, slow the development of, or cause regression of the medical condition being treated, or to alleviate to some extent the medical condition or one or more symptoms or complications of the condition. The term "therapeutically effective amount" also refers to an amount of a compound that is sufficient to elicit the biological or medical response of a cell, tissue, organ, system, animal or human that is being sought by a researcher, veterinarian, medical doctor or clinician.

[0060] The term "treating" includes alleviating, ameliorating or eliminating a medical condition or one or more symptoms or complications associated with the condition, as well as alleviating, ameliorating or eliminating one or more causes of the condition. Reference to "treating" a medical condition includes preventing the condition. The term "preventing" includes preventing, reducing the risk of developing, and delaying the onset of a medical condition or one or more symptoms or complications associated with the condition. Anti-CD79 antibody

[0061] The antibodies described herein are specific for CD79 and include all of the above forms. Antibodies can be engineered for use in specific organisms. The organism can be a human, a canine, or a commercially valuable livestock, such as a pig, a horse, a dog, a cat, a chicken, or other birds. Such engineering modifications of antibodies include, for example, CDR splicing, humanization, humanization, chimerization, or separation of human (or other organism) antibodies using any library technology or monoclonal technology known in the art.

[0062] The anti-CD79 antibody can include a heavy chain variable region selected from SEQ ID NOs: 9-17, 24-27 and 32-41, and a light chain variable region selected from SEQ ID NOs: 18-23, 28-31 and 42-56. For example, a heavy chain having a variable region of SEQ ID NO: 9 can be combined with a light chain having a variable region of SEQ ID NOs: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 71, 72, 75, 76 or 77. A heavy chain having a variable region of SEQ ID NO: 10 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO: 11 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO: 12 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO: 13 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO: 14 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO: 15 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74.A heavy chain having a variable region of SEQ ID NO: 16 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO: 17 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO: 24 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO: 25 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO: 26 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO: 27 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO:28 can be combined with a light chain having a variable region of SEQ ID NO:18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74.A heavy chain having a variable region of SEQ ID NO:33 can be combined with a light chain having a variable region of SEQ ID NO:18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO:34 can be combined with a light chain having a variable region of SEQ ID NO:18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO: 35 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO: 36 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO: 37 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO: 38 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO:39 can be combined with a light chain having a variable region of SEQ ID NO:18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74.A heavy chain having a variable region of SEQ ID NO:40 can be combined with a light chain having a variable region of SEQ ID NO:18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO:41 can be combined with a light chain having a variable region of SEQ ID NO:18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO: 71 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO: 72 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO: 75 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO: 76 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74. A heavy chain having a variable region of SEQ ID NO: 77 can be combined with a light chain having a variable region of SEQ ID NO: 18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 73 or 74.

[0063] The anti-CD79 antibody may also comprise a heavy chain variable region having 99%, 95%, 90%, 80% or 70% sequence identity to one of SEQ ID NOs: 9-17, 24-27, 32-41, 71, 72 and 75-77, and a light chain variable region selected from SEQ ID NOs: 18-23, 28-31, 42-56 and 73-74. The anti-CD79 antibody may also comprise a heavy chain variable region having 99%, 95%, 90%, 80% or 70% sequence identity to one of SEQ ID NOs: 9-17, 24-28 and 33-43, and a light chain variable region having 99%, 95%, 90%, 80% or 70% sequence identity to one of SEQ ID NOs: 18-23, 28-31, 42-56 and 73-74. The anti-CD79 antibody may also comprise a heavy chain variable region selected from SEQ ID NOs: 9-17, 24-27, 32-41, 71, 72 and 75-77, and a light chain variable region having 99%, 95%, 90%, 80% or 70% sequence identity to one of SEQ ID NOs: 18-23, 28-31, 42-56 and 73-74. The anti-CD79 antibody may bind to CD79 with an affinity of 2.0-5.1 nM, or 45 nM to 300 nM, or 2.0 to 300 nM. The anti-CD79 antibody may bind with an affinity of at least 300 nM, or at least 140 nM, or at least 100 nM, or at least 5.1 nm, at least 3.8 nM, or at least 2.4 nM.

[0064] Affinity maturation can be used with the antibodies disclosed herein to obtain anti-CD79 antibodies of desired affinity. When anti-CD79 antibodies are obtained from animals (e.g., transgenic animals carrying human antibody repertoires), antibodies prepared in transgenic animals can undergo affinity maturation. Alternatively, affinity maturation of antibodies from transgenic animals or from other technologies (e.g., display technologies) can be performed using chain shuffling methods and / or mutations of nucleic acids encoding VH and VL, followed by screening and / or selection of antibodies with greater affinity.

[0065] The most widely used method to minimize the immunogenicity of non-human antibodies while retaining specificity and affinity involves grafting the CDRs of non-human antibodies onto human frameworks that are generally selected based on their structural homology to the non-human framework (Jones et al., 1986, Nature 321:522-5; U.S. Pat. No. 5,225,539, both of which are incorporated herein by reference in their entirety). Inclusion of certain non-human residues at key positions in the framework can improve the affinity of the CDR-grafted antibodies (Bajorath et al., 1995, J Biol Chem 270:22081-4; Martin et al., 1991, Methods Enzymol. 203:121-53; Al-Lazikani, 1997, J Mol Biol 273:927-48, both of which are incorporated herein by reference in their entirety). Exemplary methods for humanizing antibodies by CDR grafting are disclosed in, for example, US Pat. No. 6,180,370, which is incorporated herein by reference in its entirety.

[0066] Improvements to the traditional CDR grafting method use a variety of hybrid selection methods, in which parts of non-human antibodies have been combined with libraries of complementary human antibody sequences for antigen binding in successive rounds of selection, during which most non-human sequences are gradually replaced by human sequences. For example, in the chain shuffling technique (Marks et al., 1992, Biotechnology 10: 779-83, which is incorporated herein by reference in its entirety), one chain of a non-human antibody is combined with a natural human library of another chain, the underlying reason being that the affinity of the non-human chain will be sufficient to constrain the selection of a human ligand (partner) to the same epitope on the antigen. The selected human ligand is then used to guide the selection of the human counterpart of the remaining non-human chain.

[0067] Other methods include chain displacement techniques, in which the non-human CDR3 is retained and only the remaining V regions (including the framework and CDR1 and 2) are replaced individually in sequential steps (e.g., U.S. Patent Application No. 20030166871; Rader et al., Proc Natl Acad Sci USA 95:8910-15, 1998; Steinberger et al., J. Biol. Chem. 275:36073-36078, 2000; Rader et al., J. Biol. Chem. 275:13668-13676, 2000, all of which are incorporated herein by reference in their entirety).

[0068] These techniques can be used to prepare antibodies suitable for use in non-human subjects by engineering CDRs into framework regions of the subject species using methods analogous to the CDR grafting methods used to prepare antibodies for use in humans.

[0069] Anti-CD79 antibodies can be prepared using a starting anti-CD79 antibody and grafting portions of the variable regions (e.g., CDRs) of the starting anti-CD79 antibody into the desired variable domain framework. The mouse variable regions are: QVQLQQSGPELVKPGASVKISCKASGYAFSYSWMNWVKQRPGKGLEWIGRIYPENGDTNYNGKFKGKVTLTADKSSSTAYMQLSSSLTSEDSAVYFCARWVYGYPHFDYWGQGTTLTVSS (heavy chain variable region, SEQ ID NO: 1) DVVMTQTPLTLSVTIGQPASISCKSSQSLLDSDGKTYLNWLLQRPGQSPKRLIYLVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPFTFGSGTKLEIK (light chain variable region, SEQ ID NO: 2)

[0070] For example, using CDRs to prepare anti-CD79 humanized antibodies: V H CDR1: YSWMN (SEQ ID NO: 3) V H CDR2:RIYPENGDTNYNGKFKG(SEQ ID NO:4) V H CDR3: WVYGYPHFDY (SEQ ID NO: 5) V L CDR1:KSSQSLLDSDGKTYLN(SEQ ID NO:6) V L CDR2: LVSKLDS (SEQ ID NO: 7) V L CDR3: WQGTHFPFT (SEQ ID NO: 8)

[0071] The three VH CDRs are placed into the framework sequence of the human heavy chain variable region to generate a VH chain: QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPENGDTNYNGKFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARWVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:9)

[0072] This VH was designated as H1. Substitutions were introduced into the framework regions of SEQ ID NO: 9 to form two additional VH chains: QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPENGDTNYNGKFKGRVTMTADTSISTAYMELSRLRSDDTAVYYCARWVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:10) SEQ ID NO: 10 was designated as H2. QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWIGRIYPENGDTNYNGKFKGRVTLTADKSISTAYMELSRLRSDDTAVYYCARWVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:11)

[0073] SEQ ID NO: 11 is designated H3. Additional changes may be introduced into SEQ ID NO: 9 in the framework region and CDR2 to increase affinity and / or stabilize the antibody against, for example, oxidation, deamination and / or protease cleavage. Examples of these heavy chain sequences are: QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPENGDTNYAGKFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARWVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:12)

[0074] SEQ ID NO: 12 was designated as H4. QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPENGDTNYAGKFKGRVTMTADTSISTAYMELSRLRSDDTAVYYCARWVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:13)

[0075] SEQ ID NO: 13 was designated as H5. QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWIGRIYPENGDTNYAGKFKGRVTLTADKSISTAYMELSRLRSDDTAVYYCARWVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:14)

[0076] SEQ ID NO: 14 was designated as H6. QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPESGDTNYAGKFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARWVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:15)

[0077] SEQ ID NO: 15 was designated as H7. QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPESGDTNYAGKFKGRVTMTADTSISTAYMELSRLRSDDTAVYYCARWVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:16)

[0078] SEQ ID NO: 16 was designated as H8. QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWIGRIYPESGDTNYAGKFKGRVTLTADKSISTAYMELSRLRSDDTAVYYCARWVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:17)

[0079] SEQ ID NO: 17 was designated as H9.

[0080] The three VL CDRs are placed into the framework sequence of the human light chain variable region to generate the VL chain: DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSDGKTYLNWFQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPFTFGGGTKVEIK(SEQ ID NO:18)

[0081] SEQ ID NO: 18 was designated as L1. Substitutions were introduced into the framework regions of SEQ ID NO: 18 to form additional VL chains: DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSDGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPFTFGGGTKVEIK(SEQ ID NO:19)

[0082] SEQ ID NO: 19 was designated as L2. DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSDGKTYLNWLQQRPGQSPKRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPFTFGGGTKVEIK(SEQ ID NO:20)

[0083] SEQ ID NO:20 is designated L3. Additional changes may be introduced into SEQ ID NO:18 in the framework regions and CDR1 to increase affinity and / or stabilize the antibody against, for example, oxidation, deamination and / or protease cleavage. Examples of these light chain sequences are: DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSSGKTYLNWFQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPFTFGGGTKVEIK(SEQ ID NO:21)

[0084] SEQ ID NO:21 was designated as L4. DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSSGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPFTFGGGTKVEIK(SEQ ID NO:22)

[0085] SEQ ID NO:22 was designated as L5. DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSSGKTYLNWLQQRPGQSPKRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPFTFGGGTKVEIK(SEQ ID NO:23)

[0086] SEQ ID NO:23 was designated as L6.

[0087] Three VH chains (H1-H3) were combined with three VL chains (L1-L3) in a combinatorial approach, and each antibody was recombinantly expressed and isolated. The H1-H3 and L1-L3 antibody chains have related framework regions that differ in several positions, which can improve affinity. The affinity of these candidates for CD79 was tested. The candidates bound to CD79 with a K range of 45nM to 300nM. d From these results, H1L2 (LB495 / PRI47, SEQ ID NOs: 9 and 19) was selected.

[0088] The present disclosure encompasses pharmaceutically acceptable salts of anti-CD79 antibodies, including those with a positive net charge, those with a negative net charge, and those with no net charge, and includes, but is not limited to, salts of anti-CD79 antibodies (including fragments thereof) as compounds in pharmaceutical compositions, in their therapeutic and diagnostic uses, and in their manufacture. Affinity Maturation

[0089] Humanized antibody H1L2 (SEQ ID NO: 9 and 19) affinity maturation. Four phage display libraries were prepared, two from heavy chain H1 (LB495), and two from light chain L2 (PRI47). Saturation mutagenesis was performed on the CDR3 of heavy chain (H1) and light chain (L2), and the mutagenized heavy chain was combined with L2, and the mutagenized light chain was combined with H1. Error-prone PCR was used for random mutagenesis of heavy chain (H1) and light chain (L2), and the mutagenized heavy chain was combined with L2, and the mutagenized light chain was combined with H1. These four libraries were produced in phage display format for further screening.

[0090] Each library was panned against CD79 in a competitive binding including humanized antibody H1L2. Clones were selected from the libraries in a competitive panning, to which humanized H1L2 antibody was added and linked to CD79 on a substrate, with which the clones had to compete. Thirty-seven (37) clones (five (5) from the CDR3 heavy chain library, four (4) from the CDR3 light chain library, eleven (11) from the heavy chain library and 17 from the light chain library) were obtained from the competitive panning of the four libraries. The sequences of these five clones from the CDR3 heavy chain library were: QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPENGDTNYNGKFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARPVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:24)

[0091] SEQ ID NO:24 was designated as LB509-A7. QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPENGDTNYNGKFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARWVYGLPHFDYWGQGTLVTVSS(SEQ ID NO:25)

[0092] SEQ ID NO:25 was designated as LB509-C2. QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPENGDTNYNGKFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARWVWGYPHFDYWGQGTLVTVSS(SEQ ID NO:26)

[0093] SEQ ID NO:26 was designated as LB509-C10. QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPENGDTNYNGKFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARWVYGYPHLDYWGQGTLVTVSS(SEQ ID NO:27)

[0094] SEQ ID NO:27 was designated as LB509-G2. QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPENGDTNYNGKFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARWVYGYPHLDYWGQGTLVTVSS(SEQ ID NO:71)

[0095] SEQ ID NO:71 is also designated as LB509-H1.

[0096] The sequences of the four clones from the CDR3 light chain library were: DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSDGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHIPFTFGGGTKVEIK(SEQ ID NO:28)

[0097] SEQ ID NO:28 was designated as LB511-A9. DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSDGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHVPFTFGGGTKVEIK(SEQ ID NO:29)

[0098] SEQ ID NO:29 was designated as LB511-B6. DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSDGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHLPFTFGGGTKVEIK(SEQ ID NO:30)

[0099] SEQ ID NO:30 was designated as LB511-F6. DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSDGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHRPFTFGGGTKVEIK(SEQ ID NO:31)

[0100] SEQ ID NO:31 was designated as LB511-F11.

[0101] The sequences of 11 clones from the heavy chain library are as follows: QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWINWVRQAPGQGLEWMGRIYPENGDTNYNGKFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARWVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:32)

[0102] SEQ ID NO:32 is named LB510 - B5. QVQLVQSGAEVKKPGASVKVSCKASGYAFGYSWMNWVRQAPGQGLEWMGRIYPENGDTNYNGKFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARWVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:33)

[0103] SEQ ID NO:33 is named LB510 - C7. QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPENGDTNYNGKFKGRVTMTRDTSISTAYMELSRLRPDDTAVYYCARWVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:34)

[0104] SEQ ID NO:34 is named LB510 - C8. QVQLVQSGAEVKKPGASVKVSCKASGYAFNYSWMNWVRQVPGQGLEWMGRIYPENGDTNYNGKFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARWVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:35)

[0105] SEQ ID NO:35 is named LB510 - F10. QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPENGDTNYNGKFKGRVTMTRDTSINTAYMELSRLRSDDTAVYYCARWVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:36)

[0106] SEQ ID NO:36 is named LB510-G4. QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWVNWVRQAPGQGLEWMGRIYPENGDTNYNGKFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARWVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:37)

[0107] SEQ ID NO:37 is named LB510-G5. QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPENGDTNYNGKFKGRVTMTRDTSISTAYMELSGLRSDDTAVYYCARWVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:38)

[0108] SEQ ID NO:38 is named LB510-G6. QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPENGDTNYNGKFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARWVYGYPHLDYWGQGTLVTVSS(SEQ ID NO:39)

[0109] SEQ ID NO:39 is named LB510-G7. QVQLVQSGAEVKKPGASVKVSCKASGYAFNYSWVNWVRQAPGQGFEWMGRIYPENGDTNYNGKFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARWVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:72)

[0110] SEQ ID NO:72 is also named LB510-H4. QVQLVQSGAEVKKPGASVKVSCKASGYAFRYSWMNWVRQAPGQGLEWMGRIYPENGGTNYNGKFKGRVTMTMDTSISTAYMELSRLRSDDTAVYYCARWVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:40)

[0111] SEQ ID NO:40 was designated as LB510-H7. QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPENGDTNYNGKFRGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARWVYGYPHFDYWGQGTLVTVSS(SEQ ID NO:41)

[0112] SEQ ID NO:41 was designated as LB510-H11.

[0113] The sequences of 17 clones from the light chain library were: DVVMTQSPLSLPVTLGLPASISCKSSQSLLDSDGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPFTFGGGTKVEIK(SEQ ID NO:42)

[0114] SEQ ID NO:42 was designated as LB512-A7. DVVMTQSPLSLPVTLGQPASISCKSSKSLLDSDGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPFTFGGGTKVEIK(SEQ ID NO:43)

[0115] SEQ ID NO:43 was designated as LB512-A10. DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSDGKTYLNWLQQRPGQSPRRIIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPFTFGGGTKVEIK(SEQ ID NO:44)

[0116] SEQ ID NO:44 was designated as LB512-B8. DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSDGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEADDVGVYYCWQGTHLPFTFGGGTKVEIK(SEQ ID NO:45)

[0117] SEQ ID NO:45 is named LB512-B10. DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSDGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGFYYCWQGTHLPFTFGGGTKVEIK(SEQ ID NO:46)

[0118] SEQ ID NO:46 is named LB512-C2. DVVMTQSPLSLPVTLGQTASISCKSSQSLLDSDGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHLPFTFGGGTKVEIK(SEQ ID NO:47)

[0119] SEQ ID NO:47 is named LB512-E2. DVVMTQSPLSLPVTLGRPASISCKSSQSLLDSGGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPFTFGGGTKVEIK(SEQ ID NO:48)

[0120] SEQ ID NO:48 is named LB512-E5. DVVMTQSPLSLPVTLGQPASISCKSSRSLLDSDGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPFTFGGGTKVEIK(SEQ ID NO:49)

[0121] SEQ ID NO:49 is named LB512-E8. DVVMTQSPLSLPVTLGLPASISCKSSQSLLDTDGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFIGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPFTFGGGTKVEIK(SEQ ID NO:50)

[0122] SEQ ID NO:50 is named LB512-E10. DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSDGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHLPFTFGGGTKVEIK(SEQ ID NO:73)

[0123] SEQ ID NO:73 is also named LB512-F7. DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSDGKTYLNWLQQRPGQPPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPFTFGGGTKVEIK(SEQ ID NO:51)

[0124] SEQ ID NO:51 is named LB512-F11. DVVMTQSPLSLPVTLGQPASITCKSSQSLLDSDGKTYLNWLQQRPGQSPRRLIYLVSKPDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPFTFGGGTKVEIK(SEQ ID NO:52)

[0125] SEQ ID NO:52 is named LB512-G2. DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSDGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHLPFTFGGGTKVEIK(SEQ ID NO:74)

[0126] SEQ ID NO:74 is also named LB512-G5. DVVMTQSPPSLPVTLGQPASISCKSSQSLLDSDGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHVPFTFGGGTKVEIK(SEQ ID NO:53)

[0127] SEQ ID NO:53 is named LB512-H5. DVVMTQSPLSLPVTLGQTASISCKSSQSLLDRDGKTYLNWLQQRPGQSPRRIIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPFTFGGGTKVEIK(SEQ ID NO:54)

[0128] SEQ ID NO:54 is named LB512-H7. DVVMTQSPLSMPVTLGLPASISCKSSQSLLDSHGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPFTFGGGTKVEIK(SEQ ID NO:55)

[0129] SEQ ID NO:55 is named LB512-H8. DVVMTQSPLSLPVTLGLPASISCKSSQSLLDSDGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHIPFTFSGGTKVEIK(SEQ ID NO:56)

[0130] SEQ ID NO:56 is named LB512-H11.

[0131] These 37 clones were screened in a competitive ELISA. Clones with better binding than the parent antibody (H1L2) were selected. Nine clones with better affinity were obtained from the competitive ELISA. Two were from the heavy chain CDR3 library (LB509-C2, SEQ ID NO:25; and LB509-G2, SEQ ID NO:27), four were from the light chain CDR3 library (LB511-A9, SEQ ID NO:28; LB511-B6, SEQ ID NO:29; LB511-F6, SEQ ID NO:30; LB511-F11, SEQ ID NO:31), two were from the heavy chain library (LB510-C7, SEQ ID NO:33; LB510-G5, SEQ ID NO:37), and one was from the light chain library (A7, SEQ ID NO:42). These clones were subjected to confirmatory dilution ELISA, and five clones that showed better binding were selected (heavy chain CDR clone LB509-C2, SEQ ID NO: 25; light chain CDR clone LB511-A9, SEQ ID NO: 28, LB511-B6, SEQ ID NO: 29, LB511-F6, SEQ ID NO: 30, and LB511-F11, SEQ ID NO: 31). Heavy chain CDR clone C2 had a change from Y to L in CDR3, while light chain clones A9, B6, F6, and F11 had an amino acid change from F to L, V, I, or R at the same position in CDR3. Two heavy chains with the H1 sequence (SEQ ID NO: 10) (named LB495), or a change from Y to L (SEQ ID NO: 25) (named LB517) were prepared, and five light chains with the L2 sequence (SEQ ID NO: 19) (named PRI47), or a change from F to I in the CDR3 of L2 (SEQ ID NO: 28) (named LB518), from F to L in the CDR3 of L2 (SEQ ID NO: 30) (named LB519), from F to R in the CDR3 of L2 (SEQ ID NO: 31) (named LB520), or from F to V in the CDR3 of L2 (SEQ ID NO: 29) (named LB521) were prepared. The two heavy chains were combined with the five light chains separately by the combination method, and the binding kinetics of these antibodies were tested.

[0132] All affinity matured antibody combinations had higher affinity for binding to CD79 than the parental antibody H1L2. d The K of affinity-matured antibodies to CD79 was 5.1 nM. d 3.6nM to 2.0nM. Anti-CD79 antibody modification

[0133] Anti-CD79 antibodies may include a method for extending the half-life (T 1 / 2 ) or / and duration of action. This portion can extend the circulating T 1 / 2 , Blood T 1 / 2 , plasma T 1 / 2 , serum T 1 / 2 , terminal T 1 / 2 、Biology 1 / 2 , eliminate T 1 / 2 or functional T 1 / 2 , or any combination thereof.

[0134] Anti-CD79 antibodies can be modified by a single part. Alternatively, anti-CD79 antibodies can be modified by two or more substantially similar or identical parts or two or more parts of the same type. Anti-CD79 antibodies can include two or more different types of parts, or different types of two or more parts. Two or more anti-CD79 antibodies can also be connected to one part. The connection between the anti-CD79 antibody and the part can be covalent or non-covalent.

[0135] The polypeptide portion may be recombinantly fused to the N-terminus or C-terminus of the heavy chain or light chain of the anti-CD79 antibody, optionally via a linker. The linker may comprise about 4-30 amino acid residues. The linker may comprise from about 6 or 8 amino acid residues to about 20 amino acid residues, or from about 6 or 8 amino acid residues to about 15 amino acid residues.

[0136] The extension portion can be human serum albumin (HSA) or a portion thereof that binds to a neonatal Fc receptor (FcRn) (e.g., domain III). HSA or its FcRn binding portion can optionally have one or more mutations that confer beneficial properties or effects. In certain embodiments, HSA or its FcRn binding portion has one or more mutations that enhance pH-dependent HSA binding to FcRn or / and increase HSA half-life, such as K573P or / and E505G / V547A. The extension portion can be an unstructured polypeptide.

[0137] The extension may be a carboxyl terminal peptide (CTP) derived from the β-subunit of human chorionic gonadotropin (hCG). In humans, the fourth, fifth, seventh and eighth serine residues of the 34 amino acid CTP of hCG-β are usually linked to O-glycans terminating in a sialic acid residue.

[0138] The extension can be 1, 2, 3, 4, 5 or more parts of a synthetic polymer. The synthetic polymer can be biodegradable or non-biodegradable. Biodegradable polymers that can be used as extensions include but are not limited to poly (2-methacryloyloxyethyl phosphorylcholine) (PMPC) and poly [oligo (ethylene glycol) methyl ether methacrylate] (POEGMA). Non-biodegradable polymers that can be used as extensions include but are not limited to poly (ethylene glycol) (PEG), polyglycerol, poly (N- (2-hydroxypropyl) methacrylamide) (PHPMA), polyoxazoline and poly (N-vinyl pyrrolidone) (PVP). The synthetic polymer can be polyethylene glycol (PEG). Pegylation can be accomplished by chemical or enzymatic, site-specific coupling or by random coupling.

[0139] The single mass (e.g., average molecular weight) or total mass of one or more synthetic polymer parts can be about 10-50, 10-20, 20-30, 30-40 or 40-50 kDa, or about 10, 20, 30, 40 or 50 kDa. The single mass (e.g., average MW) or total mass of one or more synthetic polymer parts can also be greater than about 50 kDa, such as about 50-100, 50-60, 60-70, 70-80, 80-90 or 90-100 kDa, or about 60, 70, 80, 90 or 100 kDa. Moreover, the mass (e.g., average MW) of a single synthetic polymer part can be less than about 10 kDa, such as about 1-5 or 5-10 kDa, or about 5 kDa. The single mass (e.g., average MW) or total mass of one or more synthetic polymer (e.g., PEG) parts can be about 20 or 40 kDa. Pharmaceutical composition

[0140] Other embodiments of the present disclosure relate to pharmaceutical compositions, including anti-CD79 antibodies or pharmaceutically acceptable salts, solvates or hydrates thereof, and one or more pharmaceutically acceptable excipients or carriers. The composition may optionally contain other therapeutic agents. Generally, the pharmaceutical composition comprises a therapeutically effective amount of an anti-CD79 antibody or fragment thereof, one or more pharmaceutically acceptable excipients or carriers, and optionally a therapeutically effective amount of other therapeutic agents, and is formulated for administration to a subject for therapeutic use.

[0141] Pharmaceutical compositions are generally prepared in accordance with current good manufacturing practices (GMP), for example, as recommended or required by §501(a)(2)(B) of the Federal Food, Drug, and Cosmetic Act and the International Conference on Harmonization Q7 guidelines.

[0142] Pharmaceutical compositions / preparations (formulations) can be prepared in sterile form. For example, pharmaceutical compositions / preparations for parenteral administration by injection or infusion are generally sterile. According to pharmaceutical grade sterilization standards known to those skilled in the art, such as those disclosed or required in U.S. Pharmacopoeia Chapters 797, 1072, and 1211 and 21 Code of Federal Regulations 211, sterile pharmaceutical compositions / preparations are synthesized or manufactured.

[0143] Pharmaceutically acceptable excipients and carriers include pharmaceutically acceptable substances, materials and vehicles. The limiting examples of excipient types include liquid and solid fillers, diluents, adhesives, lubricants, glidants, surfactants, dispersants, disintegrants, emulsifiers, wetting agents, suspending agents, thickeners, solvents, isotonic agents, buffers, pH regulators, absorption delay agents, stabilizers, antioxidants, preservatives, antimicrobials, antibacterial agents, antifungal agents, chelating agents, adjuvants, sweeteners, flavoring agents, coloring agents, encapsulating materials and coatings. The use of such excipients in pharmaceutical preparations / formulations is known in the art. For example, conventional vehicles and carriers include but are not limited to oils (e.g., vegetable oils, such as olive oil and sesame oil), aqueous solvents {e.g., saline, buffered saline (e.g., phosphate buffered saline [PBS]) and isotonic solutions (e.g., Ringer's solution)} and organic solvents (e.g., dimethyl sulfoxide [DMSO] and alcohols [e.g., ethanol, glycerol and propylene glycol]). Except to the extent that any conventional excipient or carrier is incompatible with the anti-CD79 antibody or fragment thereof, the present disclosure encompasses the use of conventional excipients and carriers in formulations / formulations containing the anti-CD79 antibody or fragment thereof. See, e.g., Remington: The Science and Practice of Pharmacy, 21st ed., Lippincott Williams & Wilkins (Philadelphia, Pennsylvania) (2005); Handbook of Pharmaceutical Excipients, 5th ed., Rowe et al., Eds., The Pharmaceutical Press and the American Pharmaceutical Association (2005); Handbook of Pharmaceutical Additives, 3rd ed., Ash and Ash, Eds., Gower Publishing Co. (2007); and Pharmaceutical Pre-formulation and Formulation, Gibson, Ed., CRC Press (Boca Raton, Florida) (2004).

[0144] Appropriate formulation / formulation can depend on a variety of factors, such as the route of administration selected. Potential routes of administration of pharmaceutical compositions comprising anti-CD79 antibodies or fragments thereof include, but are not limited to, oral, parenteral (including intradermal, subcutaneous, intramuscular, intravascular, intravenous, intraarterial, intraperitoneal, intramedullary, intrathecal and local), intracavitary and local (including skin / epidermis, transdermal, mucosal, transmucosal, intranasal (e.g., by nasal spray or drops), intraocular (e.g., by eye drops), pulmonary (e.g., by oral or nasal inhalation), oral, sublingual, rectal [e.g., by suppository] and vaginal [e.g., by suppository]). Local formulations / formulations can be designed to produce local or systemic therapeutic effects. In certain embodiments, anti-CD79 antibodies or fragments thereof are administered by injection (e.g., as a single dose) or infusion parenterally (e.g., intravenous, subcutaneous, intramuscular or intraperitoneal) over a period of time.

[0145] Excipients and carriers that can be used to prepare parenteral formulations include, but are not limited to, solvents (e.g., aqueous solvents such as water, saline, physiological saline, buffered saline (e.g., phosphate-buffered saline), balanced salt solutions (e.g., Ringer's BSS), and aqueous dextrose), isotonic / isotonic agents (e.g., salts [e.g., NaCl, KCl, and CaCl 2 ] and sugars [e.g., sucrose]), buffers and pH adjusters (e.g., sodium dihydrogen phosphate [monosodium phosphate] / disodium hydrogen phosphate [disodium phosphate], citric acid / sodium citrate and L-histidine / L-histidine HCl), and emulsifiers (e.g., nonionic surfactants, such as polysorbates [e.g., polysorbate 20 and 80] and poloxamers [e.g., poloxamer 188]). Protein formulations and delivery systems are discussed, for example, in AJ Banga, Therapeutic Peptides and Proteins: Formulation, Processing, and Delivery Systems, 3rd ed., CRC Press (Boca Raton, Florida) (2015).

[0146] Excipients may optionally include one or more substances that increase protein stability, increase protein solubility, inhibit protein aggregation, or reduce solution viscosity, or any combination or all thereof. Examples of such substances include, but are not limited to, hydrophilic amino acids (e.g., arginine and histidine), polyols (e.g., myo-inositol, mannitol, and sorbitol), carbohydrates {e.g., glucose (including D-glucose [dextrose]), lactose, sucrose, and trehalose}, osmotic agents (e.g., trehalose, taurine, amino acids [e.g., glycine, sarcosine, alanine, proline, serine, β-alanine, and γ-aminobutyric acid], and betaines [e.g., trimethylglycine and trimethylamine N-oxide]), and nonionic surfactants {e.g., alkyl polyglycosides, Alkyl sugars (e.g., monosaccharides [e.g., glucose] or disaccharides [e.g., maltose or sucrose] coupled to long-chain fatty acids or corresponding long-chain alcohols) and polypropylene glycol / polyethylene glycol block copolymers (e.g., poloxamers [e.g., Pluronic TM F-68] and PF-10 and its variants)}. Since such substances increase protein solubility, they can be used to increase the protein concentration in the formulation / formulation. Higher protein concentrations in the formulation / formulation are particularly advantageous for subcutaneous administration, which has limited dosing (e.g., ≤ about 1.5 mL). In addition, such substances can be used to stabilize the protein during preparation, storage, and reconstitution of lyophilized proteins.

[0147] For parenteral (e.g., intravenous, subcutaneous or intramuscular) administration, a sterile solution or suspension of an anti-CD79 antibody in an aqueous solvent containing one or more excipients can be prepared in advance and can be provided in, for example, a prefilled syringe. Alternatively, the anti-CD79 antibody can be dissolved or suspended in an aqueous solvent, which can optionally contain one or more excipients, before freeze drying (freeze-drying). Shortly before parenteral administration, a lyophilized anti-CD79 antibody stored in a suitable container (e.g., a vial) can be reconstituted with, for example, sterile water, which can optionally contain one or more excipients. If the anti-CD79 antibody is to be administered by infusion (e.g., intravenous), the reconstituted anti-CD79 antibody solution or suspension can be added to an infusion bag containing, for example, sterile saline (e.g., about 0.9% NaCl) and diluted.

[0148] Excipients that enhance transmucosal permeability of smaller proteins include, but are not limited to, cyclodextrins, alkyl saccharides (e.g., alkyl glycosides and alkyl maltosides [e.g., tetradecyl maltoside]), and bile acids (e.g., cholic acid, glycocholic acid, taurocholic acid, deoxycholic acid, deoxyglycocholic acid, chenodeoxycholic acid, and dehydrocholic acid).

[0149] Excipients that enhance the transepithelial or percutaneous permeability of smaller proteins include, but are not limited to, chemical permeation enhancers (CPEs, including fatty acids [e.g., oleic acid]), cell penetrating peptides {CPPs, including arginine-rich CPPs [e.g., polyarginine, e.g., R 6 -R 11 (For example, R 6 and R 9) and CPPs related to TAT, such as TAT ​​(49-57)] and amphiphilic CPPs [such as Pep-1 and penetrants (penetratin)]} and peptides that penetrate the skin (SPPs, such as [SPACE] peptides that penetrate the skin and enter cells). The transdermal permeability of smaller proteins can be further enhanced by using physical enhancement techniques, such as iontophoresis, cavitation or non-cavitation ultrasound, electroporation, thermal ablation, radiofrequency, microdermabrasion, microneedle or jet injection. US2007 / 0269379 provides a wide range of CPE lists. F. Milletti, Drug Discov. Today, 17: 850-860 (2012) is a review of CPPs. R. Ruan et al., Ther. Deliv., 7:89-100 (2016) discussed CPP and SPP for transdermal delivery of macromolecules, and M. Prausnitz and R. Langer, Nat. Biotechnol., 26:1261-1268 (2008) discussed various transdermal drug delivery methods.

[0150] Anti-CD79 antibodies can be delivered from a sustained release composition. As used herein, the term "slow release composition" encompasses sustained release, delayed release (prolonged-release), extended release (extended-release), slow release and controlled release compositions, systems and devices. Protein delivery systems are discussed in, for example, Banga (supra). The sustained release composition can deliver a therapeutically effective amount of anti-CD79 antibodies over an extended period of time. In certain embodiments, the sustained release composition delivers anti-CD79 antibodies in at least about 3 days, 1 week, 2 weeks, 3 weeks, 1 month (4 weeks), 6 weeks, 2 months, 3 months or longer. The sustained release composition can be administered, for example, parenterally (e.g., intravenously, subcutaneously or intramuscularly).

[0151] The sustained-release composition of protein can be in the form of, for example, a particle system, a lipid or oily composition, or an implant. The particle system includes, but is not limited to, nanoparticles, nanospheres, nanocapsules, microparticles, microspheres, and microcapsules. The nanoparticle system generally has a diameter or equivalent size less than about 1 μm. In certain embodiments, the diameter or equivalent size of the nanoparticle, nanosphere, or nanocapsule is no more than about 500, 400, or 300 nm, or no more than about 200, 150, or 100 nm. In certain embodiments, the diameter or equivalent size of the microparticle, microsphere, or microcapsule is about 1-200, 100-200, or 50-150 μm, or about 1-100, 1-50, or 50-100 μm. The nanocapsule or microcapsule usually contains a therapeutic agent in a central core, and the therapeutic agent is usually dispersed throughout the nanoparticle or microparticle or nanosphere or microsphere. In certain embodiments, the nanoparticle system is administered intravenously, while the microparticle system is administered subcutaneously or intramuscularly.

[0152] In certain embodiments, the sustained-release particle system or implant is made of a biodegradable polymer or / and a hydrogel. In certain embodiments, the biodegradable polymer includes lactic acid or / and glycolic acid [e.g., a copolymer based on L-lactic acid, such as poly (L-lactide-co-glycolide) or poly (L-lactic acid-co-D, L-2-hydroxyoctanoic acid)]. Non-limiting examples of polymers that can form hydrogels include polyvinyl alcohol, acrylate polymers (e.g., sodium polyacrylate) and other homopolymers and copolymers with relatively large amounts of hydrophilic groups (e.g., hydroxyl groups or / and carboxylate groups). The biodegradable polymer of the particle system or implant can be selected so that the polymer is substantially completely degraded at the end of the expected treatment period, and the byproducts of polymer degradation (e.g., polymers) are biocompatible.

[0153] Alternatively, the sustained-release composition of the protein can be composed of a non-biodegradable polymer. Examples of non-biodegradable polymers include, but are not limited to, poloxamers (e.g., poloxamer 407). The sustained-release composition of the protein can be composed of other natural or synthetic substances or materials, such as hydroxyapatite.

[0154] Sustained-release lipid or oily compositions of proteins can be in the form of, for example, liposomes, micelles (eg, those composed of biodegradable natural and / or synthetic polymers, such as lactosomes), and emulsions in oil.

[0155] The sustained release composition can be formulated or designed as a depot, which can be injected or implanted, for example, subcutaneously or intramuscularly. The depot can be in the form of, for example, a polymer particle system, a polymer implant, or a lipid or oily composition. The depot formulation can comprise a mixture of a protein and, for example, a biodegradable polymer [for example, poly-(lactide-co-glycolide)] or a semi-biodegradable polymer (for example, a block copolymer of lactic acid and PEG) in a biocompatible solvent system, whether or not this mixture forms a particle system or an implant.

[0156] The pharmaceutical composition can be present as a single dose in a unit dosage form, wherein all active and inactive ingredients are combined in a suitable system, and no mixing of the components is required to form the composition to be administered. The unit dosage form generally contains an effective dose of the therapeutic agent. A representative example of a unit dosage form is a single-use pen, including a pre-filled syringe, needle, and needle cover for parenteral (e.g., intravenous, subcutaneous, or intramuscular) injection of the therapeutic agent.

[0157] Alternatively, the pharmaceutical composition can be provided as a kit, wherein the therapeutic agent, excipient and carrier (e.g., solvent) are provided in two or more separate containers (e.g., ampoules, vials, tubes, bottles or syringes) and need to be combined to form the composition to be administered. The kit can include instructions for storing, preparing and administering the composition (e.g., a solution to be injected intravenously or subcutaneously).

[0158] The kit may contain all active and inactive ingredients in unit dosage form, or in two or more separate containers, and may include instructions for administering or using the pharmaceutical composition to treat a medical condition.

[0159] In certain embodiments, the kit comprises an anti-CD79 antibody or a pharmaceutical composition containing an anti-CD79 antibody, and instructions for administering or using the anti-CD79 antibody or a pharmaceutical composition containing an anti-CD79 antibody to treat an antibody-related disorder. Application of anti-CD79 antibodies

[0160] The above-mentioned anti-CD79 antibodies can be administered to subjects suffering from antibody-related disorders (e.g., diseases, disorders and / or syndromes). When the subject is a human, the anti-CD79 antibody can be a chimeric mouse-human antibody or a humanized antibody. Such chimeric or humanized antibodies are as described above. Antibody-related disorders include, for example, autoimmune diseases, certain allergies (allergies associated with antibodies), certain types of type I diabetes, etc. Autoimmune diseases that can be treated with anti-CD79 antibodies include, for example, systemic lupus erythematosus (SLE), inflammatory bowel disease (e.g., Crohn's disease and ulcerative colitis), rheumatoid arthritis, multiple sclerosis, Grave's disease, CREST syndrome, systemic sclerosis, celiac disease, achalasia, Addison's disease, adult-onset Still's disease, agammaglobulinemia, alopecia areata, amyloidosis, ankylosing spondylitis, anti-GBM / anti-TBM nephritis, antiphospholipid syndrome, autoimmune angioedema, autoimmune dysautonomia, autoimmune encephalomyelitis, autoimmune hepatitis, autoimmune inner ear disease (AIID), and ED), autoimmune myocarditis, autoimmune oophoritis, autoimmune orchitis, autoimmune pancreatitis, autoimmune retinopathy, autoimmune urticaria, axonal and neuronal neuropathy (AMAN), Baló disease, Behcet's disease, benign mucous membrane pemphigoid, bullous pemphigoid, Castleman disease (CD), celiac disease, Chagas disease, chronic inflammatory demyelinating polyneuropathy (CIDP), chronic recurrent multifocal osteomyelitis (CRMO), Churg-Strauss syndrome (CSS) or eosinophilic granulomatosis (EGPA), scar Scarring pemphigoid, Cogan's syndrome, cold agglutinin disease, congenital heart block, Coxsackie myocarditis, CREST syndrome, Crohn's disease, dermatitis herpetiformis, dermatomyositis, Devic's disease (neuromyelitis optica), discoid lupus, Dressler's syndrome, endometriosis, eosinophilic esophagitis (EoE), eosinophilic fasciitis, erythema nodosum, essential mixed cryoglobulinemia, Evans syndrome, fibromyalgia, fibrosing alveolitis, giant cell arteritis (temporal arteritis), giant cell myocarditis, glomerulonephritis, Goodpasture's syndrome, meat Granulomatosis with polyangiitis, Graves' disease, Guillain-Barre syndrome, Hashimoto's thyroiditis, hemolytic anemia, Henoch-Schonlein purpura (HSP), herpes gestationis or pemphigoid gestationis (PG), hidradenitis suppurativa (HS) (acne inversa), hypogammaglobulinemia, IgA nephropathy, IgG4-related sclerosing disease, immune thrombocytopenic purpura (ITP), inclusion body myositis (IBM), interstitial cystitis (IC), juvenile arthritis, juvenile diabetes mellitus (type 1 diabetes), juvenile myositis (JM), Kawasaki disease,Lambert-Eaton syndrome, leukocytoclastic vasculitis, lichen planus, lichen sclerosus, woody conjunctivitis, linear IgA disease (LAD), lupus, chronic Lyme disease, Meniere's disease, microscopic polyangiitis (MPA), mixed connective tissue disease (MCTD), Mooren's ulcer, Mucha-Habermann disease, multifocal motor neuropathy (MMN) or MMNCB, multiple sclerosis, myasthenia gravis, myositis, narcolepsy, neonatal lupus, neuromyelitis optica, neutropenia, ocular cicatricial pemphigoid, optic neuritis, relapsing rheumatic disease (PR), PANDAS, paraneoplastic cerebellar degeneration (PCD), paroxysmal nocturnal hemoglobinuria (PNH), Parry's Romberg syndrome, pars planitis (peripheral uveitis), Parsonage-Turner syndrome, pemphigus, peripheral neuropathy, peripheral encephalomyelitis, pernicious anemia (PA), POEMS syndrome, polyarteritis nodosa, polyglandular syndromes (type I, II, III), polymyalgia rheumatica, polymyositis, post-myocardial infarction syndrome, postpericardiotomy syndrome, primary biliary cirrhosis, primary sclerosing cholangitis, progesterone dermatitis, psoriasis, psoriatic arthritis, pure red cell agenesis (PRCA), pyoderma gangrenosum, Raynaud phenomenon, reactive arthritis, reflex sympathetic dystrophy, relapsing polychondritis, restless legs syndrome (RLS), retroperitoneal fibrosis, rheumatic fever, rheumatoid arthritis, sarcoidosis, Schmidt syndrome, scleritis, scleroderma, Syndrome, sperm and testicular autoimmunity, stiff-man syndrome (SPS), subacute bacterial endocarditis (SBE), Susac's syndrome, sympathetic ophthalmia (SO), Takayasu's arteritis, temporal arteritis / giant cell arteritis, thrombocytopenic purpura (TTP), Tolosa-Hunt syndrome (THS), transverse myelitis, type 1 diabetes, ulcerative colitis (UC), undifferentiated connective tissue disease (UCTD), uveitis, vasculitis, vitiligo, Vogt-Koyanagi-Harada disease, etc. Other antibody-related disorders that can be treated with anti-CD79 antibodies include, for example, allergies (allergies associated with antibodies), amyloidosis, certain forms of transplant rejection, etc. These and other undesirable antibody-related disorders can be treated by administering one or more anti-CD79 antibodies described herein to a subject suffering from an undesirable antibody-related disorder.

[0161] Anti-CD79 antibodies described herein can induce an anergy in the B-cells of a subject and are therefore useful for treating certain autoimmune diseases. For example, autoimmune diseases associated with anti-autoantibody responses can be treated with anti-CD79 antibodies because the induced anergy will prevent the generation of anti-autoantibodies. Anti-CD79 antibodies described herein can also be used to induce an anergy in any situation with an undesirable antibody response. Anti-CD79 antibodies described herein can be used to induce an anergy in B-cells. Anti-CD79 antibodies described herein can be used to inhibit the proliferation of B-cells.

[0162] The anti-CD79 antibodies described herein can be used to identify and / or separate B-cells in a sample and / or subject. For example, the anti-CD79 antibodies described herein can be used to diagnose B-cell malignancies or other lymphoproliferative diseases, and / or can be used as a vehicle to selectively transport agents to B-cell malignancies.

[0163] The anti-CD79 antibodies described herein can be used to directly treat B-cell malignancies, and / or can be used to construct cytotoxic T cells expressing chimeric T cell receptors (CAR-T) for treating CD79-positive B-cell malignancies. The antigen binding portion of the anti-CD79 antibodies described herein can be used as the antigen binding domain / part of CAR to prepare chimeric antigen receptors (CAR). These anti-CD79 CARs can be placed in immune cells (eg, T-cells or natural killer cells), and anti-CD79 immune cells can be used to treat diseases caused by cells expressing CD79. Such diseases include, for example, CD79-positive hematopoietic cancers (eg, lymphomas, leukemias, myeloma).

[0164] The therapeutically effective amount and frequency of administration of the anti-CD79 antibodies disclosed herein for treating antibody-related disorders, as well as the length of time for treatment with them, may depend on a variety of factors, including the nature and severity of the disorder, the efficacy of the antibody, the mode of administration, the age, weight, general health, sex and diet of the subject, and the subject's response to treatment, and may be determined by the attending physician. The therapeutically effective amount of an antibody (e.g., anti-CD79 antibody LB517 / LB519) for treating an antibody-related disorder may be from about 1, 5 or 10 mg to about 200 mg, from about 1, 5 or 10 mg to about 150 mg, from about 1, 5 or 10 mg to about 100 mg, or from about 1, 5 or 10 mg to about 50 mg, or as deemed appropriate by the attending physician, which may be administered in a single dose or divided doses. The therapeutically effective amount of the antibody can be about 1-5 mg, 5-10 mg, 10-20 mg, 20-30 mg, 30-40 mg, 40-50 mg, 50-100 mg, 100-150 mg or 150-200 mg. The therapeutically effective amount of the antibody can be about 1, 5, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 150 or 200 mg. The therapeutically effective amount of the antibody can be about 1-5 mg, 5-10 mg or 10-50 mg. The therapeutically effective amount of the antibody (e.g., anti-CD79 antibody LB517 / LB519) for treating antibody-related disorders can be about 0.01-0.1 mg / kg, 0.1-0.5 mg / kg, 0.5-1 mg / kg, 1-2 mg / kg or 2-3 mg / kg body weight, or an amount deemed appropriate by the attending physician. A therapeutically effective amount of the antibody may be about 0.01-0.1 mg / kg, 0.1-0.5 mg / kg, or 0.5-1 mg / kg body weight.

[0165] Anti-CD79 antibodies can be administered at any suitable frequency to treat antibody-related disorders. Antibodies (e.g., anti-CD79 antibodies LB517 / LB519) can be administered once a day, once every 2 days, once every 3 days, twice a week, once a week, once every 2 weeks, once every 3 weeks, once a month, once every 6 weeks, once every 2 months, or once every 3 months, or at a frequency deemed appropriate by the attending physician. Antibodies can be administered once a week or once every 2 weeks.

[0166] Similarly, anti-CD79 antibodies can be used in any suitable length of time or in any suitable total dosage to treat antibody-related conditions. Antibodies (e.g., anti-CD79 antibodies LB517 / LB519) are used in at least about 1 week, 2 weeks, 1 month (4 weeks), 6 weeks, 2 months, 3 months, 6 months, 1 year, 2 years, 3 years or longer time periods, or are considered appropriate by the attending physician. Antibody-related conditions can be chronic diseases. Chronic diseases can exist, for example, at least about 6 weeks or 2 months or longer. Antibodies can be used in a time period of at least about 6 weeks, 2 months, 3 months or 6 months. In the entire treatment regimen, 1, 2, 3, 4, 5 or 6 dosages of antibodies (e.g., anti-CD79 antibodies LB517 / LB519) can be used. In the entire treatment regimen, 1, 2 or 3 dosages of antibodies can be used.

[0167] Anti-CD79 antibodies (e.g., anti-CD79 antibodies LB517 / LB519) can also be administered in an irregular manner to treat antibody-related disorders. For example, antibodies or fragments thereof can be administered 1, 2, 3, 4, 5 or more times in an irregular manner over a period of 1 week, 2 weeks, 3 weeks, 1 month, 2 months or 3 months. In addition, anti-CD79 antibodies (e.g., anti-CD79 antibodies LB517 / LB519) can be taken when necessary (as needed) to treat antibody-related disorders. For example, antibodies can be administered 1, 2, 3, 4, 5 or more times to treat hypertension, whether in a regular or irregular manner, until blood pressure is reduced to a certain level. Once blood pressure is reduced to a certain level, it is possible to choose to stop the administration of antibodies. If blood pressure reaches or exceeds a certain level, the administration of antibodies can be resumed, whether in a regular or irregular manner. The appropriate dosage, frequency of administration and length of treatment time of the antibody can be determined by the attending physician.

[0168] In order to establish the anti-CD79 antibody of therapeutic level more quickly, at least one loading dose of antibody or its fragment can be applied before the maintenance dose. The antibody (such as anti-CD79 antibody LB517 / LB519) of loading dose can be applied, and then (i) one or more additional loading doses are applied, and then one or more therapeutically effective maintenance doses are applied, or (ii) one or more therapeutically effective maintenance doses are applied without additional loading doses, as the attending physician deems appropriate. The loading dose of the drug can be larger (for example, about 1.5, 2, 3, 4 or 5 times) than the subsequent maintenance dose, and is designed to establish the therapeutic level of the drug faster. One or more therapeutically effective maintenance doses can be any therapeutically effective amount described herein. The loading dose can be about 2 times or 3 times than the maintenance dose. The loading dose of the antibody can be applied on the 1st day, and the maintenance dose of the antibody can be applied later, for example, once a week or once every 2 weeks during the treatment. The antibody (e.g., anti-CD79 antibody LB517 / LB519) can be administered at a loading dose of about 2-10 mg, 10-20 mg, or 20-100 mg, or about 3-15 mg, 15-30 mg, or 30-150 mg on day 1, followed by a maintenance dose of about 1-5 mg, 5-10 mg, or 10-50 mg once a week or once every 2 weeks during the treatment period (e.g., at least about 2, 3, or 6 months), wherein the loading dose is approximately 2-fold or 3-fold greater than the maintenance dose, and the antibody or fragment thereof is administered parenterally (e.g., intravenously, subcutaneously, or intramuscularly).

[0169] Two (or more) loading doses of the antibody may be administered before the maintenance dose. The first loading dose of the antibody or fragment thereof may be administered on day 1, the second loading dose may be administered, for example, after about 1 or 2 weeks, and the maintenance dose may then be administered, for example, once a week or once every 2 weeks during treatment. The first loading dose may be about 3 or 4 times greater than the maintenance dose, and the second loading dose may be about 2 times greater than the maintenance dose. The antibody (e.g., anti-CD79 antibody LB517 / LB519) can be administered with a first loading dose of about 3-15 mg, 15-30 mg, or 30-150 mg, or about 4-20 mg, 20-40 mg, or 40-200 mg on day 1, a second loading dose of about 2-10 mg, 10-20 mg, or 20-100 mg about 1 or 2 weeks later, and then a maintenance dose of about 1-5 mg, 5-10 mg, or 10-50 mg once a week or once every 2 weeks during the treatment period (e.g., at least about 2, 3, or 6 months), wherein the first loading dose can be about 3 or 4 times larger than the maintenance dose, and the second loading dose can be about 2 times larger than the maintenance dose, and the antibody or fragment thereof can be administered parenterally (e.g., intravenously, subcutaneously, or intramuscularly). Combination therapy with other therapeutic agents

[0170] The present disclosure provides a method for treating an antibody-related disorder, comprising administering a therapeutically effective amount of an anti-CD79 antibody as described herein to a subject in need of treatment, optionally in combination with another therapeutic agent. The present disclosure further provides an anti-CD79 antibody as described herein, or a composition comprising an anti-CD79 antibody as described herein, which is used as a medicament, optionally in combination with another therapeutic agent. In addition, the present disclosure provides the use of an anti-CD79 antibody as described herein in the preparation of a medicament, optionally in combination with another therapeutic agent. In certain embodiments, the medicament is used to treat an antibody-related disorder.

[0171] One or more additional therapeutic agents may optionally be used in combination with an anti-CD79 antibody (e.g., anti-CD79 antibody LB517 / LB519) to treat antibody-related disorders. The optional additional therapeutic agent may be administered to a subject simultaneously with the antibody (e.g., in the same composition as the antibody or fragment thereof or in a separate composition) or sequentially (before or after) to a subject.

[0172] Optional additional therapeutic agents can be selected from immunosuppressants, anti-inflammatory agents, allergy drugs and combinations thereof. One or more immunosuppressants can be used in combination with anti-CD79 antibodies (e.g., anti-CD79 antibodies LB517 / LB519) to treat antibody-related disorders. Such immunosuppressants may include, for example, anti-CD20 antibodies (e.g., rituximab), calcineurin inhibitors (e.g., tacrolimus, cyclosporine, etc.), antiproliferative agents or IDMH inhibitors (e.g., mycophenolate mofetil, mycophenolate sodium, azathioprine, leflunomide, etc.), mTOR inhibitors (e.g., rapamycin, everolimus, etc.), steroids (e.g., corticosteroids, such as prednisone, budesonide, prednisolone, etc.) and biologics (e.g., ababatcept, adalimumab, anakinra, certolizumab, etanercept, infliximab, ixoximet, oxazolidinone ... The biologics may also include, for example, CTLA4 fusion protein, anti-TNFα antibody, IL-1 receptor antagonist protein, TNF receptor fusion protein, anti-IL17A antibody, anti-α4 integrin antibody, anti-IL6 receptor antibody, anti-p40 subunit of IL12 / IL23 antibody, anti-α 4 β 7 Integrin antibodies, anti-CD25 antibodies, and anti-CD3 antibodies.

[0173] One or more anti-inflammatory agents can be used in combination with anti-CD79 antibodies (e.g., anti-CD79 antibodies LB517 / LB519) to treat antibody-related disorders with inflammatory components. One or more anti-inflammatory agents can include, for example, inhibitors of proinflammatory cytokines or their receptors or their production (e.g., TNF-α or / and IL-6 or IL-6R). Other anti-inflammatory agents include, for example, nonsteroidal anti-inflammatory drugs (NSAIDs), immunomodulators, immunosuppressants, anti-inflammatory cytokines and compounds that increase their production, inhibitors of proinflammatory cytokines or their receptors, inhibitors of the production of proinflammatory cytokines or their receptors, inhibitors of proinflammatory transcription factors or their activation or expression, proinflammatory prostaglandins (e.g., prostaglandin E 2 [PGE 2 ]) or its receptors (e.g., EP 3) or inhibitors of their production, leukotrienes or their receptors or inhibitors of their production, inhibitors of phospholipase A2 (e.g., secreted and cytoplasmic PLA2), suppressors of C-reactive protein (CRP) activity or levels, mast cell stabilizers, phosphodiesterase inhibitors, specialized pro-resolving mediators (SPMs), other types of anti-inflammatory agents and analogs, derivatives, fragments and salts thereof.

[0174] Nonsteroidal anti-inflammatory drugs (NSAIDs) include, but are not limited to, acetic acid derivatives, fenamates, oxicams, propionic acid derivatives, salicylates, COX-2 selective inhibitors, other classes of NSAIDs, such as monoterpenoids (e.g., eucalyptol and phenols (e.g., carvacrol)), aniline pyridine carboxylic acids (e.g., clonixin), sulfonamides (e.g., nimesulide), and dual inhibitors of lipoxygenase (e.g., 5-LOX) and cyclooxygenase (e.g., COX-2) (e.g., chebulagic acid, licofelone, 2-(3,4,5-trimethoxyphenyl)-4-(N-methylindol-3-yl)thiophene, and di-tert-butylphenol-based compounds [e.g., DTPBHZ, DTPINH, DTPNHZ, and DTPSAL]); and analogs, derivatives, and salts thereof.

[0175] The glucocorticoid class of corticosteroids has anti-inflammatory effects and immunosuppressive properties. Glucocorticoids include, but are not limited to, hydrocortisone types, halogenated steroids, carbonates and their analogs, derivatives and salts.

[0176] Optional additional therapeutic agents can be used alone in any suitable manner. Potential modes of administration include, but are not limited to, oral, parenteral (including intradermal, subcutaneous, intramuscular, intravascular, intravenous, intraarterial, intraperitoneal, intramedullary, intrathecal and local), intracavitary and local (including skin / epidermis, transdermal, mucosal, transmucosal, intranasal (e.g., by nasal spray or drops), intraocular (e.g., by eye drops), pulmonary (e.g., by oral or nasal inhalation), oral, sublingual, rectal [e.g., by suppository] and vaginal [e.g., by suppository]). In certain embodiments, optional additional therapeutic agents are administered orally or parenterally (e.g., intravenously, subcutaneously or intramuscularly) alone.

[0177] One or more anti-allergic agents can be used in combination with anti-CD79 antibodies (eg, anti-CD79 antibodies LB517 / LB519) to treat antibody-related disorders. Such anti-allergic agents may include, for example, antihistamines (e.g., cetirizine, fexofenadine, levocetirizine, loratadine, bormpheniramine, chlorpheniramine, celmastine, diphenhydramine, ketotifen, naphazoline, pheniramine, desloratadine, azelastine, epinastine, olopatadine), decongestants (e.g., pseudoephedrine, phenylephrine, oxymetazoline), steroids (e.g., beclomethasone, ciclesonide, fluticasone furoate, mometasone, budesonide, triamcinolone acetonide, dexamethasone, loteprednol, prednisone epocrates), mast cell stabilizers (e.g., sodium cromolyn, lodoxamide-tromethamine, nedocromil, pemirolast) and leukotriene modifiers (e.g., monteleukast).

[0178] One or more anti-rejection drugs for transplantation can be used in combination with anti-CD79 antibodies (e.g., anti-CD79 antibodies LB517 / LB519) to treat subjects after a transplantation procedure. Such anti-rejection drugs can include, for example, calcineurin inhibitors, antiproliferative agents or IDMH inhibitors, mTOR inhibitors, and steroids.

[0179] Optional additional therapeutic agents can be used alone at any suitable frequency, including but not limited to every day (1, 2 or multiple times per day), every two or three days, twice a week, once a week, once every two weeks, once every three weeks, monthly, every two months or every three months, or in an irregular manner or as needed. The frequency of administration can depend on, for example, the selected mode of administration. The time length for treatment with optional additional therapeutic agents can be determined by the attending physician, and can be, for example, independently, at least about 1 day, 2 days, 3 days, 4 days, 1 week, 2 weeks, 3 weeks, 4 weeks (1 month), 6 weeks, 2 months, 3 months, 6 months, 1 year, 2 years, 3 years or longer. Diagnostic Applications of Anti-CD79 Antibodies

[0180] Anti-CD79 antibodies disclosed herein can also be used for the diagnosis and prognosis assessment of B-cell related diseases. In addition, such antibodies can be used to facilitate treatment decisions.

[0181] Anti-CD79 antibodies can be used to detect the presence of B-cell lineage cells in raw or processed samples obtained from a subject. Biological samples can include, for example, blood, plasma, serum, urine, cerebrospinal fluid (CSF), cells or tissues. Samples can be analyzed directly, extracted prior to analysis, or expanded in volume by adding a suitable solvent.

[0182] Biological samples can be contacted with anti-CD79 antibodies, and the samples can be screened to detect the binding of antibodies or fragments thereof to B-cells. Detection of such binding indicates the presence of B-cells in the sample. Anti-CD79 antibodies can be labeled with detectable agents (e.g., fluorescent dyes) so that the binding of antibodies to B-cells causes signals. Before the introduction of labeled anti-CD79 antibodies, B-cells in the biological sample can be fixed on a surface (direct assay), and the amount of signal equivalent to the number of labeled antibodies or fragments thereof bound to B-cells is related to the number of B-cells in the sample. B-cells in the biological sample can be captured by an unlabeled first antibody fixed on a surface, and then detected by a labeled second antibody that binds to the captured B-cells and produces a signal proportional to the number of captured B-cells (sandwich assay), wherein the unlabeled first antibody and the labeled second antibody bind to different epitopes on the B-cells, and the unlabeled first antibody or / and the labeled second antibody can independently be an anti-CD79 antibody disclosed herein.

[0183] B-cells in biological samples can be detected by competitive analysis. The sample (optionally suspended in a buffer) can be mixed with labeled anti-CD79. The resulting mixture can then be contacted with a matrix coated with a B-cell marker. The more B-cells there are in the sample, the more antibody / B-cell complexes are formed, and the less likely unbound (free) antibodies are to bind to the B-cell marker on the matrix ("competition"), and therefore the lower the signal generated.

[0184] In the above-mentioned direct, sandwich and competitive assays, the first anti-CD79 antibody can be labeled with a detectable agent. Alternatively, in direct, sandwich and competitive assays, the first anti-CD79 antibody can be unlabeled and can be bound by a labeled second antibody (e.g., an antibody that binds to the Fc region of the first antibody) after the first antibody binds to the B-cell. If the second antibody is conjugated to an enzyme, the addition of the enzyme substrate results in an enzyme / substrate reaction that produces a signal (e.g., a chromogenic, fluorescent or electrochemical signal). The absorbance, fluorescence or electrochemical signal (e.g., current) of the solid support (e.g., plate wells or beads) is measured to determine the presence and number of B-cells in the sample. Non-limiting examples of substrates for horseradish peroxidase (HRP) include 3-amino-9-ethylcarbazole (AEC), 3,3'-diaminobenzidine (DAB) and 3,3',5,5'-tetramethylbenzidine (TMB), those for alkaline phosphatase include 5-bromo-4-chloro-3-indolyl phosphate (BCIP), and those for β-glucuronidase include 5-bromo-4-chloro-3-indolyl-β-D-glucuronic acid (X-Gluc). This assay is called an enzyme-linked immunosorbent assay (ELISA).

[0185] Detectable agents include, but are not limited to, chromophores {e.g., dyes, stains, pigments, and chromophores (e.g., 3-amino-9-ethylcarbazole [AEC], 5-bromo-4-chloro-3-indolyl phosphate [BCIP], 3,3'-diaminobenzidine [DAB], and 3,3',5,5'-tetramethylbenzidine [TMB])}, fluorophores / fluorescent dyes (e.g., fluorescent dyes such as fluorescein, fluorescein isothiocyanate, and rhodamine), chemiluminescent compounds (e.g., fluorescein and luminol), radioactive isotopes (e.g., 3 H. 14 C. 32 P. 35 S and 125 I), radioactive elements (e.g., technetium), electron-dense compounds, magnetic materials and particles (e.g., paramagnetic and superparamagnetic materials and particles), magnetic resonance imaging (MRI) contrast agents (e.g., gadolinium-containing contrast agents), enzymes (e.g., horseradish peroxidase [HRP], alkaline phosphatase, luciferase, β-glucuronidase, and β-galactosidase), haptens, and toxins.

[0186] The anti-CD79 antibodies described herein can be used in a variety of immunoassays. Such assays include, but are not limited to, chromogenic, fluorescent, chemiluminescent, light scattering, radiolabeled, electrochemical, enzyme, precipitation, agglutination, coagulation, protein blot, grid blot, tissue blot, dot blot, dipstick, and biosensor assays. See, e.g., Principles and Practice of Immunoassays, C. Price and D. Newman (Eds.), Stockton Press (1997); and The Immunoassay Handbook, 2nd Ed., D. Wild (Ed.), Nature Publishing Group (2001). In addition, anti-CD79 antibodies can be used in imaging, such as by MRI.

[0187] The amount of B-cell in the detection and measurement biological sample can promote the diagnosis, prognosis and treatment of B-cell related diseases.In certain embodiments, illness is related to the B-cell level increase, and compared with the B-cell level in the corresponding sample of other experimenters without illness, the B-cell level increase in the sample of experimenter represents the disease of diagnosis experimenter.The reference B-cell level for diagnosing illness can be determined based on the B-cell level in the corresponding sample of the experimenter without disease that is numerous statistically or epidemiologically and the B-cell level in the corresponding sample of the experimenter with disease that is numerous statistically or epidemiologically.Similarly, for treatment and prognosis purposes, the B-cell level in the sample from the experimenter can be compared with the scale of the B-cell level related to the severity of B-cell related diseases, to determine the current severity of disease and predict the possible course of disease or result (for example, progress or disappearance) of disease.

[0188] In addition, detecting and measuring the number of B-cells in a biological sample can facilitate treatment decisions. In certain embodiments, based on the presence, absence, number or level of B-cells in a sample from a subject, the amount of anti-CD79 antibody administered to the subject or / and the frequency of administering the antibody to the subject are maintained or adjusted (increased or decreased), or the administration of the antibody or its fragment to the subject is stopped.

[0189] In certain embodiments, the kit comprises an anti-CD79 antibody, which may optionally be labeled with a detectable agent, and instructions for using the anti-CD79 antibody in a diagnostic application (eg, in an immunoassay). Generation of anti-CD79 antibodies

[0190] The present disclosure provides polynucleotides comprising nucleic acid sequences encoding anti-CD79 antibodies described herein. The polynucleotides may include a V sequence encoding an anti-CD79 mAb. H Domain and / or V L The polynucleotide may include a nucleic acid sequence encoding the heavy chain and / or light chain of an anti-CD79 mAb.

[0191] The disclosure further provides constructs (also referred to as expression or cloning constructs) comprising nucleic acid sequences encoding anti-CD79 antibodies as described herein. Suitable constructs include, but are not limited to, plasmids, cosmids, bacterial artificial chromosomes, yeast artificial chromosomes, lambda phages (e.g., those having deleted lysogen genes), and viruses. The construct can be free in the cell or integrated into the chromosome (either way the construct is retained and remains a construct, plasmid, and vector).

[0192] Various construct systems can be used. One type of construct utilizes DNA elements derived from animal viruses such as adenovirus, baculovirus, bovine papilloma virus, polyoma virus, SV40 virus, vaccinia virus, and retroviruses (e.g., MMTV, MOMLV, and Rous sarcoma virus). Another type of construct utilizes RNA elements derived from RNA viruses such as Eastern Equine Encephalitis Virus, Flavivirus, and Semliki Forest Virus.

[0193] In addition to V encoding for example anti-Cd79 mAb H Domain and / or V L In addition to the nucleic acid sequences of the polypeptide, polypeptide, or heavy chain, or light chain, the construct may also include various other elements for optimal expression of mRNA. For example, the construct may include a transcription promoter, a promoter plus an operator, an enhancer, an open reading frame with or without introns and / or exons, a termination signal, a splicing signal, a secretion signal sequence, or a selection marker (e.g., a gene conferring resistance to antibiotics or cytotoxic agents), or any combination or all thereof.

[0194] The disclosure also provides host cells comprising or expressing constructs encoding anti-CD79 antibodies described herein. Suitable host cells include, but are not limited to, eukaryotic cells, mammalian cells (e.g., BHK, CHO, COS, HEK293, HeLa, MDCKII, and Vero cells), insect cells (e.g., Sf9 cells), yeast cells, and bacterial cells (e.g., E. coli cells). The host cell can be a mammalian cell (e.g., a CHO cell or a HEK293 cell).

[0195] The host cell may include or express a V encoding an anti-CD79 mAb. H Domain or V L The host cell may include or express a V encoding an anti-CD79 mAb. H Domain and V L The same host cell or separate host cells may include or express V domains encoding anti-CD79 mAbs. H domain or heavy chain constructs, and V or V encoding the mAb L Individual constructs of either the structural domain or the light chain.

[0196] The construct can be transfected or introduced into the host cell by any method known in the art. Transfection agents and methods include, but are not limited to, calcium phosphate, cationic polymers (e.g., DEAE-dextran and polyethylenediamine), dendrimers, fugene, cationic liposomes, electroporation, sonoporation, cell squeezing, gene gun, viral transfection, and retroviral transduction.

[0197] Methods and conditions for culturing transfected host cells and recovering recombinantly produced anti-CD79 antibodies are known in the art and may be varied or optimized, for example, depending on the particular expression vector and / or host cell used. H Domain and / or V L Domain, or heavy chain or / and light chain. The heavy and light chains of anti-CD79 complete IgG1, IgG2 or IgG4, or the heavy and light chains of anti-CD79 Fab fragments optionally fused to extension parts are recombinantly produced.

[0198] All publications and patents cited in this specification are herein incorporated by reference as if each individual publication or patent was specifically and individually indicated to be incorporated by reference and are incorporated herein to disclose and describe the methods and / or materials in connection with the cited publication.

[0199] The following examples are intended only to illustrate the present disclosure. Other assays, studies, processes, protocols, procedures, methods, reagents and conditions may also be used alternatively as appropriate. Examples Example 1. Generation of chimeric anti-CD79 antibodies

[0200] The chimeric antibody HcLc was generated from a mouse antibody that binds to CD79 with high specificity and affinity. H Domain and V L The structural domains (SEQ ID NOs: 1 and 2) are respectively identical to those of human IgG2 C H 1. C H 2 and C H 3 domains or human κC L Domain fusion. Example 2. Generation of humanized anti-CD79 antibodies

[0201] Mouse antibodies that bind to CD79 were also humanized. The CDRs of heavy chain (IgG1) and light chain (κ) SEQ ID NO: 3-8 were grafted onto acceptor human framework sequences. Three of these CDRs (CDR-L1, CDR-L2, and CDR-H2) contained amino acid motifs (DG, DS, and NG) that may not be desirable. Different CDR-L1 was used for KSSQSLLDS S GKTYLN (SEQ ID NO: 57), two different CDR-H2 for RIYPENGDTNY A GKFKG (SEQ ID NO: 58) or RIYPE S GDTNY AGKFKG (SEQ ID NO: 59). In addition, certain framework amino acid residues of the mouse antibody are retained, including those directly adjacent to the CDR sequence or predicted to be about 100 amino acids below the CDR in the 3D immunoglobulin model. The amino acids within 30 μg / ml of the CDR are retained and may contact the antigen and support the binding of the CDR to the antigen. Approximately three mouse framework amino acid residues adjacent to the CDR are retained.

[0202] Human framework sequences were selected by aligning the mouse framework sequences with a database of human framework sequences to find the closest human homolog for each chain (usually about 65-70% sequence identity). The human VH1-2 framework was used as the human acceptor framework, which is most homologous to the mouse framework sequence of the heavy chain, and VK2-30 was used as the human acceptor framework, which is most homologous to the mouse framework sequence of the light chain. Three different VL-FR2s were used: WFQQRPGQSPRRLIY (SEQ ID NO: 60), W L QQRPGQSPRRLIY (SEQ ID NO: 61) or W L QQRPGQSP K RLIY (SEQ ID NO: 62). VH-FR1 has two amino acid changes: QVQLVQSGAEVKKPGASVKVSCKASGY A F S (SEQ ID NO:63). Three different combinations of VH-FR2 and VH-FR3 were used: VH-FR2 WVRQAPGQGLEWMG (SEQ ID NO:64) and VH-FR3 RVTMTRDTSISTAYMELSRLRSDDTAVYYCAR (SEQ ID NO:65), VH-FR2 WVRQAPGQGLEWMG (SEQ ID NO:64) and VH-FR3 RVTMTRDTSISTAYMELSRLRSDDTAVYYCAR (SEQ ID NO:65). A DTSISTAYMELSRLRSDDTAVYYCAR (SEQ ID NO: 66), and VH-FR2WVRQAPGQGLEW I G (SEQ ID NO: 67) and VH-FR3RVT L T A D K SISTAYMELSRLRSDDTAVYYCAR (SEQ ID NO: 68).

[0203] Humanized heavy chains H1-H3 (SEQ ID NO: 9-11) and L1-L3 (SEQ ID NO: 18-20) were prepared and mixed and matched using a combinatorial approach to construct a humanized clone library (H1-H3 and L1-L3 were mixed using a combinatorial approach). Each member was isolated and tested for affinity binding to CD79. The binding data obtained are listed in Table 1 below: Load Sample ID KD(M) kon(1 / Ms) kdis(1 / s) Full X^2 Full R^2 H1L2 4.5E-08 9.1E+04 4.1E-03 0.0208 0.9973 H2L2 4.6E-08 1.0E+05 4.6E-03 0.0315 0.9943 H1L3 4.9E-08 8.2E+04 4.0E-03 0.0264 0.9966 H1L1 5.9E-08 9.2E+04 5.4E-03 0.0408 0.9922 HkDJ 6.0E-08 9.3E+04 5.6E-03 0.0833 0.9857 H2L3 6.1E-08 8.4E+04 5.1E-03 0.0679 0.9869 H2L1 9.0E-08 7.9E+04 7.1E-03 0.0444 0.9895 H3L3 1.3E-07 7.9E+04 1.1E-02 0.1020 0.9611 H3L2 1.4E-07 6.1E+04 8.6E-03 0.1402 0.9726 H3L1 2.9E-07 4.3E+04 1.3E-02 0.1566 0.9472 Table 1. Binding affinity of humanized antibodies

[0204] The lead antibody H1L2 (LB495 / PRI47, SEQ ID NOs: 9 and 19) was selected. Example 3. Affinity maturation of humanized antibody H1L2 (LB495 / PRI47, SEQ ID NOs: 9 and 19)

[0205] Humanized antibody H1L2 (SEQ ID NO: 9 and 19) affinity maturation. Four phage display libraries were prepared, two from heavy chain H1 (SEQ ID NO: 10), and two from light chain L2 (SEQ ID NO: 20). Saturation mutagenesis was performed on CDR3 to prepare CDR3 heavy chain and CDR3 light chain libraries, and error-prone PCR was also used for random mutagenesis of each of H1 heavy chain and L2 light chain to prepare heavy chain library and light chain library. Each heavy chain library was combined with parental light chain L2 (SEQ ID NO: 19), and each light chain library was paired with parental heavy chain H1 (SEQ ID NO: 9) to prepare phage display libraries for candidates for affinity maturation. Each library was panned for CD79 in a competitive analysis with parental H1L2 antibody, and the affinity of the binding clones obtained in the panning was tested in ELISA. A VH clone with a mutation in CDR3 (SEQ ID NO: 25) was selected, and a VL clone with a mutation at the same position of CDR3 (SEQ ID NO: 28-31) was selected. These heavy and light chains were reformatted into full-length IgGs, which were produced by 293 cells, purified, and then tested for binding. The binding data for these full-length IgGs are listed in Table 2 below: Load Sample ID KD(M) Kon(1 / Ms) Kdis(1 / s) Full X^2 Full R^2 PRI43 / PRI47 5.10E-09 8.41E+04 4.29E-04 0.1219 0.9989 LB517 / PRI47 3.55E-09 8.75E+04 3.11E-04 0.1046 0.9991 LB495 / 518 3.64E-09 9.12E+04 3.32E-04 0.0725 0.9994 LB495 / 519 3.14E-09 8.03E+04 2.52E-04 0.1107 0.9989 LB495 / 520 3.76E-09 9.96E+04 3.74E-04 0.0723 0.9993 LB495 / 521 3.65E-09 9.91E+04 3.62E-04 0.0502 0.9993 LB517 / 518 2.24E-09 8.85E+04 1.99E-04 0.0533 0.9994 LB517 / 519 2.00E-09 7.97E+04 1.59E-04 0.0834 0.9994 LB517 / 520 2.40E-09 9.78E+04 2.34E-04 0.0720 0.9993 LB517 / 521 2.30E-09 8.75E+04 2.01E-04 0.0736 0.9992 Table 2. Binding affinities of affinity matured antibodies

[0206] PRI 47 is light chain L2 (SEQ ID NO: 19), PRI 43 and LB495 are heavy chains H1 (SEQ ID NO: 9), LB517 is heavy chain LB509-C2 (SEQ ID NO: 25), LB518 is light chain LB511-A9 (SEQ ID NO: 28), LB519 is light chain LB511-F6 (SEQ ID NO: 30), LB520 is light chain LB511-F11 (SEQ ID NO: 31), and LB521 is light chain LB511-B6 (SEQ ID NO: 29).

[0207] Affinity matured antibodies exhibit higher affinity than the parental humanized antibody. Example 4. Development of Antibody Cell Lines

[0208] CHO cells (Invitrogen) were cultured in serum-free medium (CD FortiCHO, Invitrogen) and co-transfected with separate plasmids encoding the heavy and light chains of the anti-CD79 antibody using Freestyle Max transfection reagent (Invitrogen). Prior to transfection, the antibody expression plasmids were linearized by restriction digestion with Sea I. Antibody expression in conditioned medium was measured by ELISA.

[0209] Transfected cells were stably selected for 2 weeks with 10 μg / ml puromycin and 500 μg / ml G418. After single cell cloning, high-producing CHO cell clones were screened and isolated. Antibody production in shake flask cultures was estimated to be >500 mg / L. Example 5. Recombinant production of humanized anti-CD79 antibodies

[0210] HEK293F cells (Invitrogen) were cultured in serum-free medium and co-transfected with plasmids encoding LB517 and LB519 or plasmids expressing other antibody variants. On day 5 after transfection, cell culture supernatants were harvested and subjected to protein A chromatography to purify antibodies. Example 6. Human CD79a / 79b knock-in mice

[0211] C57BL / 6 embryonic stem cells were engineered to replace the mouse CD79a and CD79b loci with nucleic acids encoding human CD79a and human CD79b. The engineered C57BL / 6 ES cells were implanted into female C57BL / 6 mice to obtain human CD79a / b positive C57BL / 6 offspring. The expression of human CD79a and CD79b on B-cells was confirmed using FACS with anti-human CD79 antibodies. Example 7. Anti-CD79 antibody-induced B-cell anergy in human CD79 C57BL / 6 mice

[0212] Human CD79 knock-in mice were treated with humanized anti-CD79 antibodies LB517 / LB519 for 18 hours prior to the assay. Then, ex vivo RBC-lysed splenocytes (1E6 / 100 μL) were stained with anti-B220 and fluorescently labeled anti-hCD79 or PTEN or B cell receptors. LB517 / LB519 (hCur14 FALA) is able to bind to B cells and compete with Curly-14 ( Figure 3a ). The intracellular levels of PTEN and cell surface BCR were also characterized by staining with appropriate antibodies. LB517 / LB519 was able to induce PTEN expression ( Figure 3b ) and downregulate BCR expression ( Figure 3c ).

[0213] The calcium flux of the B-cells of these mice was tested after activation by the B-cell receptor. RBC lysed spleen cells (1E7 / mL) were stained with anti-B220 (B-cells) and loaded with a calcium-sensitive dye (Indo-1 AM) for 1 hour before flow analysis. Changes in intracellular Ca2+ levels were measured in FORTESSA (BD Bioscience) running Flow-Jo software (Tree Star), and the fluorescence emission at 405nm and 485nm was recorded. Baseline calcium was obtained for 30 seconds before adding the stimulus to 100μL culture medium. The basal calcium measurements were subtracted from the AUCs after stimulation to generate acute stimulation quantification. LB517 / LB519 stimulated calcium influx ( Figure 4a ) and desensitizes B-cell receptors ( Figure 4b ).

[0214] B-cells from C57BL / 6 mice were also tested for tyrosine phosphorylation following B-cell receptor stimulation. These studies showed that pretreatment of B-cells with humanized anti-CD79 antibodies LB517 / LB519 desensitized B-cells to activation through the B-cell receptor because tyrosine phosphorylation was inhibited in anti-CD79 antibody-treated B-cells. Example 8. Treatment of Type 1 Diabetes

[0215] Mice expressing the IgM heavy chain transgenes VH125 and VH281 on a NOD and C57BL / 6 background were obtained. VH125.C57BL / 6 mice were backcrossed to C57BL / 6-H2g7 (JAX) to generate VH125.C57BL / 6.H2g7. Two consecutive blood glucose readings > 250 mg / dL (one touch) confirmed that the VH125.C57BL / 6.H2g7 mice were diabetic.

[0216] Prediabetic (serial blood glucose readings >150-<200 mg / dL) VH125NOD mice were treated with anti-CD79 antibody or saline. Figure 1 It was shown that mice treated with normal saline developed type I diabetes (80% by week 5). Figure 1 It was also shown that mice treated with anti-CD79 antibody developed less type I diabetes (25-30% by week 5). Example 9. Treatment of type I diabetes with humanized anti-CD79 antibody LB517 / LB519

[0217] C57BL / 6 mice expressing human CD79a / b in their B-cells were prepared according to Example 6. hCD79a / b C57BL / 6 mice have a PTPn22 R620W knock-in ROSA26 locus with an intervening floxed stop cassette. Cre of the PTPn22 R620W autoimmune risk allele induced by tamoxifen tam Driven expression of β-cells, combined with streptozotocin (STZ) treatment to destroy pancreatic β cells, activated the disease in adult mice.

[0218] Six to eight week old mice were injected (ip) with 40 mg / kg of STZ (Sigma-Aldrich) for 4 consecutive days. Blood glucose levels were measured twice a week using a Bayer Contour Meter (Bayer) starting 2 weeks after the last STZ injection. Diabetes was defined by an increase in glucose levels >500 mg / dL in two consecutive tests.

[0219] PTPn22 R620W hCD79a / b C57BL / 6 mice were treated with humanized anti-CD79 antibodies LB517 / LB519 or saline before / during / after treatment of mice with STZ. Mice were monitored by blood glucose levels twice a week. Diabetes was defined by an increase in glucose levels >500 mg / dL on two consecutive tests. Example 10. Treatment of arthritis

[0220] On day 0, C57BL / 6 mice were immunized with bovine or chicken type II collagen (CII) emulsified in complete Freund's adjuvant (CFA). 21 days later, mice were immunized a second time with CII emulsified in incomplete Freund's adjuvant (IFA). 1 mg of anti-CD79 or isotype control immunoglobulin was administered subcutaneously (sc) on day 0. Two hours after mAb injection, mice were immunized with collagen. Clinical scores were assessed after the secondary immunization of each paw, using a score range of 0 to 4, as previously described (Hardy, 2014). Anti-CD79 significantly inhibited the development of arthritis ( Figure 5 ).

[0221] hCD79a / b C57BL / 6 mice expressing human CD79a / b in their B-cells were prepared according to Example 6. hCD79a / b C57BL / 6 mice were immunized with bovine or chicken type II collagen (CII) emulsified in Freund's complete adjuvant (CFA) on day 0. 21 days later, mice were secondary immunized with CII emulsified in Freund's incomplete adjuvant (IFA).

[0222] Anti-mouse CD79 D265A, anti-human CD79 (humanized anti-CD79 antibody LB517 / LB519), anti-CD20 (18B12), or isotype control immunoglobulins will be administered subcutaneously (sc) on day 0. Two hours after mAb injection, mice will be immunized with collagen.

[0223] Clinical scores will be assessed after the second immunization of each paw using a score range of 0 to 4 as previously described (Hardy, 2014). Example 11. Treatment of systemic lupus

[0224] MRL / lpr,MRL / lpr-Thy1.1 mice were injected with 0.5mg anti-CD79 weekly for 6-17 weeks. Anti-CD79 reduced inflammation in the kidneys and mandibular salivary glands and improved survival at 17 weeks qwk ( Figure 6 ).

[0225] hCD79a / b C57BL / 6 mice expressing human CD79a / b in their B-cells were prepared according to Example 6. hCD79a / b C57BL / 6 mice were treated with acute tamoxifen to delete SH2-containing inositol lipid phosphatase (SHIP-1) and / or inositol lipid phosphatase PTEN in B-cells. Getahun et al., J Exp Med. 2016 May 2; 213(5):751-69, which is incorporated herein by reference in its entirety for all purposes.

[0226] In tamoxifen-treated human CD79 C57BL6 mice, humanized anti-CD79 antibodies LB517 / LB519, anti-CD20 (18B12), or isotype control immunoglobulin will be administered subcutaneously once a week starting at 8 weeks of age (after the appearance of autoantibodies). Anti-chromatin autoantibody production, glomerular deposition, and mouse health will be monitored. Example 12. Epitope Mapping of Humanized Anti-CD79 Antibodies

[0227] The binding epitope of the anti-CD79 antibody was located by competition and structural analysis of various fragments of the CD79 antigen. Example 13. Construction of anti-CD79 CAR-T for cancer treatment

[0228] The anti-CD79 antibody L1H2 was converted into a scFv antibody and fused with the transmembrane domain 4-1BB and the intracellular domain of CD3zeta in the vector LB586. Plasmid LB586 was transfected into CHO cells and appropriate antibiotics were selected for 2 weeks. Stable cells were stained with soluble biotinylated CD79 antigen and then conjugated with streptavidin PE. The binding of CD79 antigen was confirmed by flow cytometry analysis ( Figure 2 ). The nucleic acid sequence of the anti-CD79 chimeric antigen receptor is:

[0229] The amino acid sequence of the anti-CD79 chimeric antigen receptor is: DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSDGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKSRVEAEDVGVYYCWQGTHLPFTFGGGTKVEIKGGGGSGG GGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPENGDTNYNGKFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARWVYGLPHFDYWGQ GTLVTVSSASTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGG CELRVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR(SEQ ID NO:70)

[0230] Anti-CD79 CAR-T vectors are used to generate cytotoxic T-cells to treat CD79-positive cancers. Example 14. Treatment of multiple sclerosis

[0231] C57BL6 / J mice were immunized with 200 μg of myelin oligodendrocyte glycoprotein (MOG) 35-55 emulsified in 200 μL of complete Freund's adjuvant (CFA) according to standard procedures. The emulsifier was injected subcutaneously at two sites, followed by two intraperitoneal (ip) injections of 200 ng pertussis toxin (PTX) in phosphate buffered saline (PBS), the first 1-2 hours after MOG35-55 and the second 24 hours thereafter. 1 mg of anti-CD79 was administered weekly starting on day 19. EAE scores and body weight were assessed daily to assess the severity and stage of the disease. Treatment with anti-CD79 slowed the progression of the disease model ( Figure 7 ). Example 15 Engineering of affinity matured humanized antibody LB517 / 519

[0232] There are 2 "NG" motifs in VH CDR2 (SEQ ID NO:4), which are associated with high Asn deamidation propensity leading to complications. Asn residues can be mutated to eliminate the risk of deamidation if the mutation does not negatively affect antigen interaction. The first NG motif was engineered to NS (SEQ ID NO:75) and cloned into expression vector LB630. The second NG motif was engineered to NA (SEQ ID NO:76) and cloned into expression vector LB631. Both NG motifs were engineered (SEQ ID NO:77) and cloned into expression vector LB632. QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPESGDTNYNGKFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARWVYGLPHFDYWGQGTLVTVSS(SEQ ID NO:75)

[0233] SEQ ID NO:75 was designated as LB630. QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPENGDTNYAGKFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARWVYGLPHFDYWGQGTLVTVSS(SEQ ID NO:76)

[0234] SEQ ID NO:76 was designated as LB631. QVQLVQSGAEVKKPGASVKVSCKASGYAFSYSWMNWVRQAPGQGLEWMGRIYPESGDTNYAGKFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARWVYGLPHFDYWGQGTLVTVSS(SEQ ID NO:77)

[0235] SEQ ID NO:77 was designated as LB632.

[0236] After pairing with the light chain expression vector LB519, the engineered antibodies were produced and purified from 293 cells by transient transfection. Antigen binding kinetics were characterized and presented in Table 3 below: Load Sample ID KD(M) Kon(1 / Ms) Kdis(1 / s) Full X^2 Full R^2 PRI43 / PRI47 9.22E-09 4.17E+04 3.84E-04 0.8803 0.9934 LB517 / 519 7.06E-10 5.87E+04 4.14E-05 0.6118 0.9964 LB630 / 519 <1.0E-12 5.58E+04 <1.0E-07 0.6271 0.9972 LB631 / 519 1.04E-09 5.86E+04 6.10E-05 0.3757 0.9982 LB632 / 519 8.37E-10 6.10E+04 5.11E-05 0.3287 0.9983 Table 3. Binding affinity of affinity matured antibodies

[0237] Among the 3 engineered antibodies, LB631 / 519 and LB632 / 519 had slightly less effective binding compared to LB517 / 519, whereas antigen binding of LB630 / 519 was greatly improved.

[0238] The biological activities of these engineered antibodies were characterized similarly to those in Example 7. Compared to LB517 / 519, the engineered antibodies exhibited similar B-cell desensitization activity ( FIGS. 3 and 4 ).

[0239] It should be understood that, although specific embodiments have been illustrated and described, various modifications may be made thereto and are contemplated herein. It should also be understood that the present disclosure is not limited to the specific examples provided herein. The description and illustration of the embodiments and examples of the present disclosure herein are not intended to be interpreted in a restrictive sense. It should also be understood that all aspects of the present disclosure are not limited to the specific description, construction or relative proportions set forth herein, which may depend on different conditions and variables. Various modifications and changes in the form and details of the embodiments and examples of the present disclosure will be apparent to those skilled in the art. Therefore, it is contemplated that the present disclosure also encompasses any and all such modifications, changes and equivalents.

Claims

1. Use of an anti-CD79 antibody in the preparation of a medicament for administration to a human subject suffering from rheumatoid arthritis, in, The anti-CD79 antibody comprises a heavy chain and a light chain, wherein the variable region of the heavy chain has at least 95% sequence identity with a sequence selected from the group consisting of SEQ ID NOs: 9-17, 24-27, 32-41, 71, 72 and 75-77, and the variable region of the light chain has at least 95% sequence identity with the sequence of SEQ ID NO:

30.

2. The use according to claim 1, in, The variable region of the heavy chain has at least 99% sequence identity with a sequence selected from the group consisting of SEQ ID NOs: 9-17, 24-27, 32-41, 71, 72 and 75-77, and the variable region of the light chain has at least 99% sequence identity with the sequence of SEQ ID NO:

30.

3. The use according to claim 1, in, The sequence of the variable region of the heavy chain is SEQ ID NO:25, and the sequence of the variable region of the light chain is SEQ ID NO:

30.

4. The use according to claim 1, in, The sequence of the variable region of the heavy chain is SEQ ID NO:75, and the sequence of the variable region of the light chain is SEQ ID NO:

30.

5. The use according to claim 1, in, The sequence of the variable region of the heavy chain is SEQ ID NO:76, and the sequence of the variable region of the light chain is SEQ ID NO:

30.

6. The use according to claim 1, in, The sequence of the variable region of the heavy chain is SEQ ID NO:77, and the sequence of the variable region of the light chain is SEQ ID NO:

30.

7. Use of an anti-CD79 antibody in the preparation of a medicament for administration to a human subject suffering from systemic lupus, in, The anti-CD79 antibody comprises a heavy chain and a light chain, wherein the variable region of the heavy chain has at least 95% sequence identity with a sequence selected from the group consisting of SEQ ID NOs: 9-17, 24-27, 32-41, 71, 72 and 75-77, and the variable region of the light chain has at least 95% sequence identity with the sequence of SEQ ID NO:

30.

8. The use according to claim 7, in, The variable region of the heavy chain has at least 99% sequence identity with a sequence selected from the group consisting of SEQ ID NOs: 9-17, 24-27, 32-41, 71, 72 and 75-77, and the variable region of the light chain has at least 99% sequence identity with the sequence of SEQ ID NO:

30.

9. The use according to claim 7, in, The sequence of the variable region of the heavy chain is SEQ ID NO:25, and the sequence of the variable region of the light chain is SEQ ID NO:

30.

10. The use according to claim 7, in, The sequence of the variable region of the heavy chain is SEQ ID NO:75, and the sequence of the variable region of the light chain is SEQ ID NO:

30.

11. The use according to claim 7, in, The sequence of the variable region of the heavy chain is SEQ ID NO:76, and the sequence of the variable region of the light chain is SEQ ID NO:

30.

12. The use according to claim 7, in, The sequence of the variable region of the heavy chain is SEQ ID NO:77, and the sequence of the variable region of the light chain is SEQ ID NO:

30.

13. Use of an anti-CD79 antibody in the preparation of a medicament for administration to a human subject suffering from type 1 diabetes, in, The anti-CD79 antibody comprises a heavy chain and a light chain, wherein the variable region of the heavy chain has at least 95% sequence identity with a sequence selected from the group consisting of SEQ ID NOs: 9-17, 24-27, 32-41, 71, 72 and 75-77, and the variable region of the light chain has at least 95% sequence identity with the sequence of SEQ ID NO:

30.

14. The use according to claim 13, in, The variable region of the heavy chain has at least 99% sequence identity with a sequence selected from the group consisting of SEQ ID NOs: 9-17, 24-27, 32-41, 71, 72 and 75-77, and the variable region of the light chain has at least 99% sequence identity with the sequence of SEQ ID NO:

30.

15. The use according to claim 13, in, The sequence of the variable region of the heavy chain is SEQ ID NO:25, and the sequence of the variable region of the light chain is SEQ ID NO:

30.

16. The use according to claim 13, in, The sequence of the variable region of the heavy chain is SEQ ID NO:75, and the sequence of the variable region of the light chain is SEQ ID NO:

30.

17. The use according to claim 13, in, The sequence of the variable region of the heavy chain is SEQ ID NO:76, and the sequence of the variable region of the light chain is SEQ ID NO:

30.

18. The use according to claim 13, in, The sequence of the variable region of the heavy chain is SEQ ID NO:77, and the sequence of the variable region of the light chain is SEQ ID NO:

30.

19. Use of an anti-CD79 antibody in the preparation of a medicament for administration to a human subject suffering from multiple sclerosis, wherein the anti-CD79 antibody comprises a heavy chain and a light chain, wherein the variable region of the heavy chain has at least 95% sequence identity with a sequence selected from the group consisting of SEQ ID NO: 9-17, 24-27, 32-41, 71, 72 and 75-77, and the variable region of the light chain has at least 95% sequence identity with the sequence of SEQ ID NO:

30.

20. The use according to claim 19, in, The variable region of the heavy chain has at least 99% sequence identity with a sequence selected from the group consisting of SEQ ID NOs: 9-17, 24-27, 32-41, 71, 72 and 75-77, and the variable region of the light chain has at least 99% sequence identity with the sequence of SEQ ID NO:

30.

21. The use according to claim 19, in, The sequence of the variable region of the heavy chain is SEQ ID NO:25, and the sequence of the variable region of the light chain is SEQ ID NO:

30.

22. The use according to claim 19, in, The sequence of the variable region of the heavy chain is SEQ ID NO:75, and the sequence of the variable region of the light chain is SEQ ID NO:

30.

23. The use according to claim 19, in, The sequence of the variable region of the heavy chain is SEQ ID NO:76, and the sequence of the variable region of the light chain is SEQ ID NO:

30.

24. The use according to claim 19, in, The sequence of the variable region of the heavy chain is SEQ ID NO:77, and the sequence of the variable region of the light chain is SEQ ID NO:30.

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